An outbreak of multidrug-resistant, quinolone-resistant Salmonella enterica serotype typhimurium DT104

DEFF Research Database (Denmark)

Molbak, K.; Baggesen, Dorte Lau; Aarestrup, Frank Møller

1999-01-01

Background Food-borne salmonella infections have become a major problem in industrialized countries. The strain of Salmonella enterica serotype typhimurium known as definitive phage type 104 (DT104) is usually resistant to five drugs: ampicillin, chloramphenicol, streptomycin, sulfonamides......, and tetracycline. An increasing proportion of DT104 isolates also have reduced susceptibility to fluoroquinolones. Methods The Danish salmonella surveillance program determines the phage types of all typhimurium strains from the food chain, and in the case of suspected outbreaks, five-drug-resistant strains...... are characterized by molecular methods. All patients infected with five-drug-resistant typhimurium are interviewed to obtain clinical and epidemiologic data. In 1998, an outbreak of salmonella occurred, in which the strain of typhimurium DT104 was new to Denmark. We investigated this outbreak and report our...

Isolation of Salmonella enterica subsp. enterica (O:4,5:i and Salmonella enterica subsp. Typhimurium from free-living domestic pigeons (Columba livia

Directory of Open Access Journals (Sweden)

R.C. Rocha-e-Silva

2014-10-01

Full Text Available The present study reports the isolation of Salmonella enterica in organs of free-living domestic pigeons. In the clinic examination, the presence of feces in the peri-cloacal and abdominal regions were observed, as well as symptoms such as cachexy, incoordination and opisthotonos. Before any therapeutic protocol was applied the bird died and a necropsy was then performed for the removal of spleen, liver, kidney and intestine for bacteriological examination and antibiotic sensitivity test. Salmonella enterica subsp.enterica (O:4,5:i- and Salmonella enterica subsp. enterica serovar Typhimurium were isolated from the liver and intestine and the sensitivity test demonstrated that these strains are sensitive to several antibiotics.

Detection of Salmonella enterica Serovar Typhimurium from Avians Using Multiplex-PCR

Directory of Open Access Journals (Sweden)

Alireza Talebi

2011-09-01

Full Text Available Abstract Salmonella enterica serovar Typhimurium and S.enterica serovar Enteritidis are the most frequently isolated serovars from food-borne diseases throughout the world. According to their antigenic profiles, salmonella shows different disease syndromes and host specificities. It is necessary and important to discriminate salmonella serovars from each other in order to ensure that each pathogen and its epidemiology are correctly recognized. Many PCR-based methods have been developed to identify salmonella serovars. The objective of present study was to identify S. Typhimurium in avians from different regions including: North, Northwest and capital city (Tehran of Iran. Also in this research, the quality of CHROMagar™ Salmonella medium (CAS medium in veterinary medicine was evaluated. The results of present study showed that out of 1870 intestine samples, fifty two S. Typhimurium including broiler (n=13, layer (n=12, duck (n=5, goose (n=5, sparrow (n=8, canary (n=3, pigeon (n=5 and African grey parrot (n=1 were identified using serotyping as well as multiplex-PCR. In conclusion, important measures must be taken on prevention and propagation of S. Typhimurium among avians. CHROMagar™ Salmonella medium has high levels of sensitivity and specificity and reduced the time to final identification of salmonella spp. in comparison with biochemical tests.

Defining the Core Genome of Salmonella enterica Serovar Typhimurium for Genomic Surveillance and Epidemiological Typing

Science.gov (United States)

Fu, Songzhe; Octavia, Sophie; Tanaka, Mark M.; Sintchenko, Vitali

2015-01-01

Salmonella enterica serovar Typhimurium is the most common Salmonella serovar causing foodborne infections in Australia and many other countries. Twenty-one S. Typhimurium strains from Salmonella reference collection A (SARA) were analyzed using Illumina high-throughput genome sequencing. Single nucleotide polymorphisms (SNPs) in 21 SARA strains ranged from 46 to 11,916 SNPs, with an average of 1,577 SNPs per strain. Together with 47 strains selected from publicly available S. Typhimurium genomes, the S. Typhimurium core genes (STCG) were determined. The STCG consist of 3,846 genes, a set that is much larger than that of the 2,882 Salmonella core genes (SCG) found previously. The STCG together with 1,576 core intergenic regions (IGRs) were defined as the S. Typhimurium core genome. Using 93 S. Typhimurium genomes from 13 epidemiologically confirmed community outbreaks, we demonstrated that typing based on the S. Typhimurium core genome (STCG plus core IGRs) provides superior resolution and higher discriminatory power than that based on SCG for outbreak investigation and molecular epidemiology of S. Typhimurium. STCG and STCG plus core IGR typing achieved 100% separation of all outbreaks compared to that of SCG typing, which failed to separate isolates from two outbreaks from background isolates. Defining the S. Typhimurium core genome allows standardization of genes/regions to be used for high-resolution epidemiological typing and genomic surveillance of S. Typhimurium. PMID:26019201

Sequential acquisition of R-plasmids in vivo by Salmonella typhimurium.

Science.gov (United States)

Platt, D J; Sommerville, J S; Gribben, J

1984-01-01

Salmonella typhimurium, resistant only to trimethoprim and sulphamethoxazole, was isolated from the faeces and blood of a chronic alcoholic patient in acute renal failure. The isolates harboured an 18 Md non-conjugative plasmid. He was dialysed peritoneally and treated with ampicillin; four days later there was no clinical improvement and his peritoneal dialysis fluid (PDF) had become infected. Salm. typhimurium was isolated from faeces and PDF. Both isolates were additionally resistant to ampicillin and contained two plasmids (55 Md and 18 Md). Therapy was changed to chloramphenicol and gentamicin was added to the PDF. Two weeks later Salm. typhimurium was again isolated from PDF and faeces. The PDF isolate was unchanged but 4% of the colonies isolated from this faecal specimen were resistant to chloramphenicol and had acquired an additional 62 Md plasmid. From all PDF and faecal specimens two different strains of Escherichia coli and one strain of Klebsiella pneumoniae were isolated which contained plasmids indistinguishable, on the basis of molecular weight and transferable resistance markers, from those acquired by Salm. typhimurium. The transferability of these plasmids in vitro to E. coli K12 and to the patient's initial Salm. typhimurium was studied and the results discussed.

Plasma-treated polyethylene film: A smart material applied for Salmonella Typhimurium detection

International Nuclear Information System (INIS)

Peng-Ubol, Triranat; Phinyocheep, Pranee; Daniel, Philippe; Panbangred, Watanalai; Pilard, Jean-François; Thouand, Gerald; Durand-Thouand, Marie-José

2012-01-01

Salmonella is a major cause of foodborne illness worldwide and is not allowed to be present in any food in all countries. The purpose of this study is to develop a simple alternative method for the detection of Salmonella based on functionalized polyethylene (PE) surfaces. Salmonella Typhimurium was used as a model bacterium. PE film was treated using dielectric plasma in order to alter the wettability of the PE surface and consequently introduce functionality on the surface. The PE film characterized by ATR-FTIR spectroscopy revealed the presence of C=O stretching of ketones, aldehydes and carboxylic acids. The antibodies against O or H antigens of Salmonella and S. Typhimurium were then respectively immobilized on the PE surface after activation of the carboxylic group using NHS/EDC followed by protein A. The evidences from ATR-FTIR, scanning electron microscopy and optical microscopy showed the presence of S. Typhimurium attached to the plasma treated PE surfaces via the two types of anti-Salmonella antibody. The plasma treated PE film developed is simple and allows efficient association of bacterial cells on the treated surfaces without the necessity of time-consuming centrifugation and washing steps for isolation of the cells. This material is considered to be a smart material applicable for S. Typhimurium detection. Highlights: ► We developed a functionalized polyethylene film for bacterial detection. ► We modified the surface of polyethylene film by plasma treatment. ► ATR-FTIR spectroscopy was used to analyze the functionality on the PE surface. ► We introduced Salmonella Typhimurium on the modified PE film. ► SEM revealed the presence of S. Typhimurium on the plasma treated PE film.

Phage typing or CRISPR typing for epidemiological surveillance of Salmonella Typhimurium?

Science.gov (United States)

Mohammed, Manal

2017-11-07

Salmonella Typhimurium is the most dominant Salmonella serovar around the world. It is associated with foodborne gastroenteritis outbreaks but has recently been associated with invasive illness and deaths. Characterization of S. Typhimurium is therefore very crucial for epidemiological surveillance. Phage typing has been used for decades for subtyping of S. Typhimurium to determine the epidemiological relation among isolates. Recent studies however have suggested that high throughput clustered regular interspaced short palindromic repeats (CRISPR) typing has the potential to replace phage typing. This study aimed to determine the efficacy of high-throughput CRISPR typing over conventional phage typing in epidemiological surveillance and outbreak investigation of S. Typhimurium. In silico analysis of whole genome sequences (WGS) of well-documented phage types of S. Typhimurium reveals the presence of different CRISPR type among strains belong to the same phage type. Furthermore, different phage types of S. Typhimurium share identical CRISPR type. Interestingly, identical spacers were detected among outbreak and non-outbreak associated DT8 strains of S. Typhimurium. Therefore, CRISPR typing is not useful for the epidemiological surveillance and outbreak investigation of S. Typhimurium and phage typing, until it is replaced by WGS, is still the gold standard method for epidemiological surveillance of S. Typhimurium.

Plasma-treated polyethylene film: A smart material applied for Salmonella Typhimurium detection

Energy Technology Data Exchange (ETDEWEB)

Peng-Ubol, Triranat [Department of Chemistry, Faculty of Science, Mahidol University, Rama 6 Rd, Phayathai, Bangkok 10400 (Thailand); Phinyocheep, Pranee, E-mail: scppo@mahidol.ac.th [Department of Chemistry, Faculty of Science, Mahidol University, Rama 6 Rd, Phayathai, Bangkok 10400 (Thailand); Daniel, Philippe [Laboratoire de Physique de l' Etat Condense (LPEC-UMR CNRS 6087), Universite du Maine, Avenue Olivier Messiaen, 72085, Le Mans Cedex 9 (France); Panbangred, Watanalai [Department of Biotechnology and Mahidol University-Osaka University Collaborative Research Center for Bioscience and Biotechnology (MU-OU: CRC), Faculty of Science, Mahidol University, Rama 6 Rd, Phayathai, Bangkok 10400 (Thailand); Pilard, Jean-Francois [Unite de Chimie Organique Moleculaire et Macromoleculaire (UCO2M-UMR CNRS 6011), Universite du Maine, Avenue Olivier Messiaen, 72085 Le Mans Cedex 9 (France); Thouand, Gerald; Durand-Thouand, Marie-Jose [Genie des Procedes Environnement et Agroalimentaire (GEPEA UMR CNRS 6144), Departement Genie Biologique, IUT de la Roche/Yon, Universite de Nantes, 18 Bd G. Defferre, 85035 La Roche sur Yon (France)

2012-12-01

Salmonella is a major cause of foodborne illness worldwide and is not allowed to be present in any food in all countries. The purpose of this study is to develop a simple alternative method for the detection of Salmonella based on functionalized polyethylene (PE) surfaces. Salmonella Typhimurium was used as a model bacterium. PE film was treated using dielectric plasma in order to alter the wettability of the PE surface and consequently introduce functionality on the surface. The PE film characterized by ATR-FTIR spectroscopy revealed the presence of C=O stretching of ketones, aldehydes and carboxylic acids. The antibodies against O or H antigens of Salmonella and S. Typhimurium were then respectively immobilized on the PE surface after activation of the carboxylic group using NHS/EDC followed by protein A. The evidences from ATR-FTIR, scanning electron microscopy and optical microscopy showed the presence of S. Typhimurium attached to the plasma treated PE surfaces via the two types of anti-Salmonella antibody. The plasma treated PE film developed is simple and allows efficient association of bacterial cells on the treated surfaces without the necessity of time-consuming centrifugation and washing steps for isolation of the cells. This material is considered to be a smart material applicable for S. Typhimurium detection. Highlights: Black-Right-Pointing-Pointer We developed a functionalized polyethylene film for bacterial detection. Black-Right-Pointing-Pointer We modified the surface of polyethylene film by plasma treatment. Black-Right-Pointing-Pointer ATR-FTIR spectroscopy was used to analyze the functionality on the PE surface. Black-Right-Pointing-Pointer We introduced Salmonella Typhimurium on the modified PE film. Black-Right-Pointing-Pointer SEM revealed the presence of S. Typhimurium on the plasma treated PE film.

Phenotypic and genotypic characterization of Salmonella Typhimurium variant 5- isolates associated with an outbreak of food-borne disease in Paz de Rio, Boyacá, Colombia, in 2010 = Caracterización fenotípica y genotípica de Salmonella Typhimurium variante 5- asociada a un brote de enfermedad transmitida por alimentos en el municipio de Paz de Río, Boyacá, 2010 = Caracterización fenotípica y genotípica de Salmonella Typhimurium variante 5- asociada a un brote de enfermedad transmitida por alimentos en el municipio de Paz de Río, Boyacá, 2010

Directory of Open Access Journals (Sweden)

Díaz Osorio, Miguel Ángel

2014-01-01

Full Text Available Introduction: Salmonella enterica serotype Typhimuri-um variant 5- is a pathogen closely related to animals, especially pigeons, which has been also associ- ated in rare cases with sporadic infections in humans. However, epidemiological surveillance systems have enabled the detection of this variant in human out- breaks. Objective: To characterize by means of phenotypic and genotypic techniques the isolates of Salmonella Typhimurium variant 5- associated with an outbreak of food-borne disease in Paz de Rio, Boyacá, Colombia (2010, in order to establish their molecular relationships. Materials and methods: Twelve isolates of Salmonella -spp., were analyzed by biochemical, serotyping and antimicrobial susceptibility tests. Pulsed-field gel electrophoresis (PFGE with Xball Blnl enzymes was used to establish their molecular relationships Results: All isolates were confirmed as Salmonella spp. They were resistant to tetracycline and streptomycin and sensitive to the rest of antibiotics tested. Eleven isolates were identified as Salmonella Typhimurium variant 5- and grouped in COIN10.JPX.X01.0168 pattern using the enzyme Xball two isolates in this group were confirmed using the enzyme Blnl with the COIN10.JPX.A26.0002 pattern. One isolate was identified as Salmonella Typhimurium with COIN10.JPX.X01.0221 pattern with the enzyme Xbal Conclusion: This is the first outbreak in Colombia of foodborne illness epidemiologically associated with isolates of Typhimurium variant 5 -, which Epidemiologic Sur were phenotypically and genetically related.

Acid environments affect biofilm formation and gene expression in isolates of Salmonella enterica Typhimurium DT104.

Science.gov (United States)

O'Leary, Denis; McCabe, Evonne M; McCusker, Matthew P; Martins, Marta; Fanning, Séamus; Duffy, Geraldine

2015-08-03

The aim of this study was to examine the survival and potential virulence of biofilm-forming Salmonella Typhimurium DT104 under mild acid conditions. Salmonella Typhimurium DT104 employs an acid tolerance response (ATR) allowing it to adapt to acidic environments. The threat that these acid adapted cells pose to food safety could be enhanced if they also produce biofilms in acidic conditions. The cells were acid-adapted by culturing them in 1% glucose and their ability to form biofilms on stainless steel and on the surface of Luria Bertani (LB) broth at pH7 and pH5 was examined. Plate counts were performed to examine cell survival. RNA was isolated from cells to examine changes in the expression of genes associated with virulence, invasion, biofilm formation and global gene regulation in response to acid stress. Of the 4 isolates that were examined only one (1481) that produced a rigid biofilm in LB broth at pH7 also formed this same structure at pH5. This indicated that the lactic acid severely impeded the biofilm producing capabilities of the other isolates examined under these conditions. Isolate 1481 also had higher expression of genes associated with virulence (hilA) and invasion (invA) with a 24.34-fold and 13.68-fold increase in relative gene expression respectively at pH5 compared to pH7. Although genes associated with biofilm formation had increased expression in response to acid stress for all the isolates this only resulted in the formation of a biofilm by isolate 1481. This suggests that in addition to the range of genes associated with biofilm production at neutral pH, there are genes whose protein products specifically aid in biofilm production in acidic environments. Furthermore, it highlights the potential for the use of lactic acid for the inhibition of Salmonella biofilms. Copyright © 2015 Elsevier B.V. All rights reserved.

A comparison of cecal colonization of Salmonella enterica serotype Typhimurium in white leghorn chicks and Salmonella-resistant mice

Directory of Open Access Journals (Sweden)

Bogomolnaya Lydia M

2008-10-01

Full Text Available Abstract Background Salmonellosis is one of the most important bacterial food borne illnesses worldwide. A major source of infection for humans is consumption of chicken or egg products that have been contaminated with Salmonella enterica serotype Typhimurium, however our knowledge regarding colonization and persistence factors in the chicken is small. Results We compared intestinal and systemic colonization of 1-week-old White Leghorn chicks and Salmonella-resistant CBA/J mice during infection with Salmonella enterica serotype Typhimurium ATCC14028, one of the most commonly studied isolates. We also studied the distribution of wild type serotype Typhimurium ATCC14028 and an isogenic invA mutant during competitive infection in the cecum of 1-week-old White Leghorn chicks and 8-week-old CBA/J mice. We found that although the systemic levels of serotype Typhimurium in both infected animal models are low, infected mice have significant splenomegaly beginning at 15 days post infection. In the intestinal tract itself, the cecal contents are the major site for recovery of serotype Typhimurium in the cecum of 1-week-old chicks and Salmonella-resistant mice. Additionally we show that only a small minority of Salmonellae are intracellular in the cecal epithelium of both infected animal models, and while SPI-1 is important for successful infection in the murine model, it is important for association with the cecal epithelium of 1-week-old chicks. Finally, we show that in chicks infected with serotype Typhimurium at 1 week of age, the level of fecal shedding of this organism does not reflect the level of cecal colonization as it does in murine models. Conclusion In our study, we highlight important differences in systemic and intestinal colonization levels between chick and murine serotype Typhimurium infections, and provide evidence that suggests that the role of SPI-1 may not be the same during colonization of both animal models.

Development of an Immunomagnetic Separation Method for Viable Salmonella Typhimurium Detected by Flow Cytometry

DEFF Research Database (Denmark)

Ahmed, Shakil; Rubahn, Horst-Günter; Erdmann, Helmut

2016-01-01

for detection of food-related bacteria. In this study, a flow cytometry based immunomagnetic separation (IMS) method for the isolation and enrichment of Salmonella Typhimurium from liquid samples was developed and optimized. Both polyclonal and monoclonal antibodies have been used to couple with 1 micron sized...... and bacteria, immunocapture time, staining and buffering conditions for the viability assays were optimized. The capture efficiency of IMS was>98% for a range of Salmonella Typhimurium cell concentrations from 103 to 105/mL using 108/mL bead concentration. The method proved to have high (98%) specificity...

Genome and Transcriptome Adaptation Accompanying Emergence of the Definitive Type 2 Host-Restricted Salmonella enterica Serovar Typhimurium Pathovar

OpenAIRE

Kingsley, Robert A.; Kay, Sally; Connor, Thomas; Barquist, Lars; Sait, Leanne; Holt, Kathryn E.; Sivaraman, Karthi; Wileman, Thomas; Goulding, David; Clare, Simon; Hale, Christine; Seshasayee, Aswin; Harris, Simon; Thomson, Nicholas R.; Gardner, Paul

2013-01-01

Salmonella enterica serovar Typhimurium definitive type 2 (DT2) is host restricted to Columba livia (rock or feral pigeon) but is also closely related to S. Typhimurium isolates that circulate in livestock and cause a zoonosis characterized by gastroenteritis in humans. DT2 isolates formed a distinct phylogenetic cluster within S. Typhimurium based on whole-genome-sequence polymorphisms. Comparative genome analysis of DT2 94-213 and S. Typhimurium SL1344, DT104, and D23580 identified few diff...

Genomics of an emerging clone of Salmonella serovar Typhimurium ST313 from Nigeria and the Democratic Republic of Congo.

Science.gov (United States)

Leekitcharoenphon, Pimlapas; Friis, Carsten; Zankari, Ea; Svendsen, Christina Aaby; Price, Lance B; Rahmani, Maral; Herrero-Fresno, Ana; Fashae, Kayode; Vandenberg, Olivier; Aarestrup, Frank M; Hendriksen, Rene S

2013-10-15

Salmonella enterica serovar Typhimurium ST313 is an invasive and phylogenetically distinct lineage present in sub-Saharan Africa. We report the presence of S. Typhimurium ST313 from patients in the Democratic Republic of Congo and Nigeria. Eighteen S. Typhimurium ST313 isolates were characterized by antimicrobial susceptibility testing, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing (MLST). Additionally, six of the isolates were characterized by whole genome sequence typing (WGST). The presence of a putative virulence determinant was examined in 177 Salmonella isolates belonging to 57 different serovars. All S. Typhimurium ST313 isolates harbored resistant genes encoded by blaTEM1b, catA1, strA/B, sul1, and dfrA1. Additionally, aac(6')1aa gene was detected. Phylogenetic analyses revealed close genetic relationships among Congolese and Nigerian isolates from both blood and stool. Comparative genomic analyses identified a putative virulence fragment (ST313-TD) unique to S. Typhimurium ST313 and S. Dublin. We showed in a limited number of isolates that S. Typhimurium ST313 is a prevalent sequence-type causing gastrointestinal diseases and septicemia in patients from Nigeria and DRC. We found three distinct phylogenetic clusters based on the origin of isolation suggesting some spatial evolution. Comparative genomics showed an interesting putative virulence fragment (ST313-TD) unique to S. Typhimurium ST313 and invasive S. Dublin.

Comparação dos caldos selenito cistina, tetrationato Muller-Kauffmann e Rappaport-Vassiliadis no isolamento de Salmonella Typhimurium

Directory of Open Access Journals (Sweden)

L.G. Ávila

2012-06-01

Full Text Available Three selective enrichment broths - selenite cystine (SC, Muller-Kauffmann tetrathionate (MKT and Rappaport-Vassiliadis (RV - were compared, for Salmonella Typhimurium isolation from rectal swabs of a calf experimentally infected. The bacteriological procedure involved pre-enrichment in Hajna-GN broth (only for the samples inoculated in RV broth, selective enrichment (SC, MKT and RV broths, culture in modified brilliant green agar (BGA, presumptive biochemistry tests (using triple-sugar-iron agar and lysine-agar and slide agglutination test with poli-O and poli-H Salmonella antisera. SC and MKT broths were more efficient in the isolation of Salmonella Typhimurium (12 positive samples, whereas RV broth had a lower efficiency in the microbiological isolation (ten positive samples.

House Sparrows Do Not Constitute a Significant Salmonella Typhimurium Reservoir across Urban Gradients in Flanders, Belgium.

Science.gov (United States)

Rouffaer, Lieze Oscar; Lens, Luc; Haesendonck, Roel; Teyssier, Aimeric; Hudin, Noraine Salleh; Strubbe, Diederik; Haesebrouck, Freddy; Pasmans, Frank; Martel, An

2016-01-01

In recent decades major declines in urban house sparrow (Passer domesticus) populations have been observed in north-western European cities, whereas suburban and rural house sparrow populations have remained relatively stable or are recovering from previous declines. Differential exposure to avian pathogens known to cause epidemics in house sparrows may in part explain this spatial pattern of declines. Here we investigate the potential effect of urbanization on the development of a bacterial pathogen reservoir in free-ranging house sparrows. This was achieved by comparing the prevalence of Salmonella enterica subspecies enterica serotype Typhimurium in 364 apparently healthy house sparrows captured in urban, suburban and rural regions across Flanders, Belgium between September 2013 and March 2014. In addition 12 dead birds, received from bird rescue centers, were necropsied. The apparent absence of Salmonella Typhimurium in fecal samples of healthy birds, and the identification of only one house sparrow seropositive for Salmonella spp., suggests that during the winter of 2013-2014 these birds did not represent any considerable Salmonella Typhimurium reservoir in Belgium and thus may be considered naïve hosts, susceptible to clinical infection. This susceptibility is demonstrated by the isolation of two different Salmonella Typhimurium strains from two of the deceased house sparrows: one DT99, typically associated with disease in pigeons, and one DT195, previously associated with a passerine decline. The apparent absence (prevalence: house sparrows and the association of infection with clinical disease suggests that the impact of Salmonella Typhimurium on house sparrows is largely driven by the risk of exogenous exposure to pathogenic Salmonella Typhimurium strains. However, no inference could be made on a causal relationship between Salmonella infection and the observed house sparrow population declines.

Polisakarida Mengandung Mannan dari Bungkil Inti Sawit Sebagai Antimikroba Salmonella typhimurium pada Ayam

Directory of Open Access Journals (Sweden)

M. Tafsin

2007-08-01

Full Text Available Mannan containing polysaccharides could be used as an alternative to replace antibiotics due to their capacity to block the colonization of pathogenic bacteria in the intestine of poultry. The aim of this study was to investigate mannan containing polysaccharides from palm kernel meal (PKM and its inhibitory effect against Salmonella typhimurium. Hot water extractions were used to isolate mannan containing polysaccharides from cell wall of PKM. In vivo studies were conducted using broiler and layer chicks that were challenged orally with 104 cfu Salmonella typhimurium on third day. Split plot design was used as experimental design with strain as main plot and level of mannan polysaccharides as sub plot. The levels of mannan containing polysaccharides that were used consisted of 0 (R0; 1000 (R1; 2000 (R2; 3000 (R3; 4000 (R4 ppm, in term of total sugar. The results indicated that compared to the control group, feeding PKM containing mannan 4000 ppm decreased (P<0.01 Salmonella typhimurium incidence. The addition of mannan did not affect feed consumption. On the contrary, the addition of 4000 ppm mannan gave significantly higher feed/weight gain ratio of the chicks (P<0.05. The administration of feed supplemented with mannan from PKM did not influence weight gain of poultry. It is concluded that mannan from PKM can prevent the colonization of Salmonella typhimurium in poultry.

Characterization of a Salmonella typhimurium mutant defective in phosphoribosylpyrophosphate synthetase

DEFF Research Database (Denmark)

Jochimsen, Bjarne; Hove-Jensen, Bjarne; Garber, Bruce B.

1985-01-01

This study describes the isolation and characterization of a mutant (strain GP122) of Salmonella typhimurium with a partial deficiency of phosphoribosylpyrophosphate (PRPP) synthetase activity. This strain was isolated in a purE deoD gpt purine auxotroph by a procedure designed to select guanosin...

Prevalence and antimicrobial susceptibility of salmonellae isolates from reptiles in Taiwan.

Science.gov (United States)

Chen, Chun-Yu; Chen, Wan-Ching; Chin, Shih-Chien; Lai, Yen-Hsueh; Tung, Kwong-Chung; Chiou, Chien-Shun; Hsu, Yuan-Man; Chang, Chao-Chin

2010-01-01

Pets, including reptiles, have been shown to be a source of Salmonella infection in humans. Due to increasing popularity and variety of exotic reptiles as pets in recent years, more human clinical cases of reptile-associated Salmonella infection have been identified. However, limited information is available with regard to serotypes in different reptiles (turtles, snakes, and lizards) and antimicrobial resistance of Salmonella in pet reptiles. The current study was thus conducted to determine the prevalence of Salmonella colonization in pet reptiles. Salmonella organisms were isolated from 30.9% of 476 reptiles investigated. The isolation prevalences were 69.7% (23/33), 62.8% (27/43), and 24.3% (97/400) in snakes, lizards, and turtles, respectively. A total of 44 different Salmonella serovars were identified. Compared with S. Heron, Bredeney, Treforest, and 4,[5],12:i:-, S. Typhimurium isolates were resistant to many antimicrobials tested, and notably 61.1% of the isolates were resistant to cephalothin. The results indicated that raising reptiles as pets could be a possible source of Salmonella infection in humans, particularly zoonotic Salmonella serovars such as S. Typhimurium that may be resistant to antimicrobials.

Persistence of a Salmonella enterica serovar typhimurium DT12 clone in a piggery and in agricultural soil amended with Salmonella-contaminated slurry

DEFF Research Database (Denmark)

Baloda, Suraj B.; Christensen, Lise; Trajcevska, Silvija

2001-01-01

Prevalence of Salmonella enterica on a Danish pig farm presenting recurrent infections was investigated. A comparison of the pulsed-held gel electrophoresis patterns of fecal isolates from piggeries, waste slurry, and agricultural soil amended with Salmonella-contaminated animal waste (slurry......) and subclinical isolates from the same farm (collected in 1996 and later) showed identical patterns, indicating long-term persistence of the Salmonella enterica serovar Typhimurium DT12 clone in the herd environment. Furthermore, when Salmonella-contaminated slurry was disposed of on the agricultural soil (a...... common waste disposal practice), the pathogen was isolated up to 14 days after the spread, indicating potentially high risks of transmission of the pathogen in the environment, animals, and humans....

Prevalence and antibiotic resistance of Salmonella Enteritidis and Salmonella Typhimurium in raw chicken meat at retail markets in Malaysia.

Science.gov (United States)

Thung, T Y; Mahyudin, N A; Basri, D F; Wan Mohamed Radzi, C W J; Nakaguchi, Y; Nishibuchi, M; Radu, S

2016-08-01

Salmonellosis is one of the major food-borne diseases in many countries. This study was carried out to determine the occurrence of Salmonella spp., Salmonella Enteritidis, and Salmonella Typhimurium in raw chicken meat from wet markets and hypermarkets in Selangor, as well as to determine the antibiotic susceptibility profile of S. Enteritidis and S. Typhimurium. The most probable number (MPN) in combination with multiplex polymerase chain reaction (mPCR) method was used to quantify the Salmonella spp., S. Enteritidis, and S. Typhimurium in the samples. The occurrence of Salmonella spp., S. Enteritidis, and S. Typhimurium in 120 chicken meat samples were 20.80%, 6.70%, and 2.50%, respectively with estimated quantity varying from retail chicken meat could be a source of multiple antimicrobial-resistance Salmonella and may constitute a public health concern in Malaysia. © 2016 Poultry Science Association Inc.

Characterization of Salmonella enterica Serovar Typhimurium DT104 Isolated from Denmark and Comparison with Isolates from Europe and the United States

DEFF Research Database (Denmark)

Baggesen, Dorte Lau; Sandvang, D.; Aarestrup, Frank Møller

2000-01-01

A total of 136 isolates of Salmonella enterica serovar Typhimurium DT104 from Denmark (n = 93), Germany (n = 10), Italy (n = 4), Spain (n = 5), and the United Kingdom (n = 9) were characterized by antimicrobial resistance analysis, plasmid profiling, pulsed-field gel electrophoresis (PFGE......) with the restriction enzymes XbaI and BlnI, and analysis for the presence of integrons and antibiotic resistance genes. The isolates from Denmark were from nine pig herds, while the isolates from other countries were both of animal and of human origin. All but 10 isolates were resistant to ampicillin, chloramphenicol......, spectinomycin, streptomycin, sulfonamides, and tetracycline. Five isolates from the United Kingdom and Spain were sensitive to all antibiotics examined, whereas four isolates from the United Kingdom and the United States were also resistant to one or more of the antibiotics, namely, gentamicin, neomycin...

Antimicrobial resistance in Salmonella enterica subsp. enterica serovar typhimurium from humans and production animals

DEFF Research Database (Denmark)

Seyfarth, Anne Mette; Wegener, Henrik Caspar; FrimodtMoller, N.

1997-01-01

: Poultry strains were usually resistant only to ampicillin, white pig and cattle isolates were most often resistant to sulphonamide, tetracycline and streptomycin. Typing of the strains showed that some animal strains and human strains were indistinguishable. In conclusion, while antimicrobial resistance......We have studied the frequency of antimicrobial resistance and epidemiological relatedness among 473 isolates of Salmonella enterica subsp, enterica serovar typhimurium (S. typhimurium) from human and veterinary sources. The human strains were clinical isolates from patients with diarrhoea sent...... to the State Serum Institute during August 1993 (228 isolates). The animal strains were isolated from clinical or subclinical infections in cattle (48 isolates), pigs (99 isolates) or poultry (98 isolates), all from 1993. All strains were tested against 22 different antimicrobial agents used in both human...

Salmonella Typhimurium infection in the porcine intestine

DEFF Research Database (Denmark)

Schauser, Kirsten; Olsen, John Elmerdahl; Larsson, Lars-Inge

2005-01-01

The normal intestinal epithelium is renewed with a turnover rate of 3-5 days. During Salmonella infection increased cell loss is observed, possibly as a result of programmed cell death (PCD). We have, therefore, studied the effects of Salmonella Typhimurium infection on three elements involved...... in scattered epithelial cells and the number of positive cells increased with increasing times of exposure to Salmonella (P

Human isolates of Salmonella enterica serovar Typhimurium from Taiwan displayed significantly higher levels of antimicrobial resistance than those from Denmark.

Science.gov (United States)

Torpdahl, Mia; Lauderdale, Tsai-Ling; Liang, Shiu-Yun; Li, Ishien; Wei, Sung-Hsi; Chiou, Chien-Shun

2013-02-01

Salmonella enterica serovar Typhimurium is a major zoonotic pathogen with a high prevalence of antimicrobial resistance. This pathogen can disseminate across borders and spread far distances via the food trade and international travel. In this study, we compared the genotypes and antimicrobial resistance of 378 S. Typhimurium isolates collected in Taiwan and Denmark between 2009 and 2010. Genotyping revealed that many S. Typhimurium strains were concurrently circulating in Taiwan, Denmark and other countries in 2009 and 2010. When compared to the isolates collected from Denmark, the isolates from Taiwan displayed a significantly higher level of resistance to 11 of the 12 tested antimicrobials. Seven genetic clusters (A-G) were designated for the isolates. A high percentage of the isolates in genetic clusters C, F and G were multidrug-resistant. Of the isolates in cluster C, 79.2% were ASSuT-resistant, characterized by resistance to ampicillin, streptomycin, sulfamethoxazole, and tetracycline. In cluster F, 84.1% of the isolates were ACSSuT-resistant (resistant to ASSuT and chloramphenicol). Cluster G was unique to Taiwan and characterized in most isolates by the absence of three VNTRs (ST20, ST30 and STTR6) as well as a variety of multidrug resistance profiles. This cluster exhibited very high to extremely high levels of resistance to several first-line drugs, and among the seven clusters, it displayed the highest levels of resistance to cefotaxime and ceftazidime, ciprofloxacin and gentamicin. The high prevalence of antimicrobial resistance in S. Typhimurium from Taiwan highlights the necessity to strictly regulate the use of antimicrobials in the agriculture and human health care sectors. Copyright © 2012 Elsevier B.V. All rights reserved.

Simultaneous oral administration of Salmonella Infantis and S. Typhimurium in chicks.

Science.gov (United States)

Murakami, Koichi; Maeda-Mitani, Eriko; Onozuka, Daisuke; Noda, Tamie; Sera, Nobuyuki; Kimura, Hirokazu; Fujimoto, Shuji; Murakami, Satoshi

2017-01-01

To confirm the hypothesis that Salmonella enterica subspecies enterica serovar ( S. ) Infantis has higher basic reproductive rates in chicks compared with other Salmonella serovars, 1-day-old specific-pathogen-free chicks ( n  = 8) were challenged simultaneously with S. Infantis and S. Typhimurium per os . Challenged chicks (Group A) were then housed with non-infected chicks (Group B, n  = 4) for 6 days (from 2 to 8 days of age). Group B birds were then housed with other non-infected birds (Group C, n  = 4), which were then transferred to cages containing a further group of untreated chicks (Group D, n  = 2). A control group consisting of four non-infected chicks was used for comparison. All chickens were humanely sacrificed at 18 days of age, and Salmonella from bowel and liver samples were enumerated. Both serovars were isolated from all groups except the control group. S. Typhimurium was isolated at a greater frequency than S. Infantis from the bowel samples of chicks from Groups B, C and D, while no differences in colonisation rates were observed between the two serovars in liver samples from Groups B, C and D. S. Typhimurium, but not S. Infantis, was immunohistochemically detected in the lamina propria of the cecum and rectum in five birds of Group A. Despite the competitive administration, neither of the two serovars completely excluded the other, and no differences were observed in basic reproductive rates between the two serovars. These findings, together with data from previous studies, suggest that the initial quantitative domination of S. Infantis in chicken flocks may explain why this serovar is predominant in broiler chickens.

Isolation and identification of Salmonella typhimurium from raw cow, sheep and goat milk in Chahamaha Va Bakhteyari Province

Directory of Open Access Journals (Sweden)

F Tajbakhsh

2014-05-01

Full Text Available Salmonella typhimurium and S. enteritidisare known as the major causes of food-borne infection throughout the world. The present study was carried out to investigate the prevalence of S. typhimurium in raw milks of Chahamaha Va Bakhteyari province. For this reason, a total of 550 raw milks (consisting of 200 cow, 175 sheep and 175 goat milk samples were collected through October 2011 to March 2012 from dairy herds around Shahrekord. The samples were cultured and the isolated colonies were confirmed by PCR using species-specific ST11 and ST15 primers. According to the results, a total of 20 samples (3.63% were found positive for Salmonellaspp.Amongst, 14 (2.54% of cow milk, 2 (0.36% of sheep milk and 4 (0.72% of goat milk samples were contaminated. Using PCR, 9 (1.63% samples were contaminated with S. typhimurium. The results indicated a relatively high occurrence of S. typhimurium in raw milks. Therefore, it is essential to maintain hygienic measures during milking and handling. Besides, it is recommended not to use raw milk for the manufacturing of dairy products such as cheese and ice-cream.

Genome and transcriptome adaptation accompanying emergence of the definitive type 2 host-restricted Salmonella enterica serovar Typhimurium pathovar.

Science.gov (United States)

Kingsley, Robert A; Kay, Sally; Connor, Thomas; Barquist, Lars; Sait, Leanne; Holt, Kathryn E; Sivaraman, Karthi; Wileman, Thomas; Goulding, David; Clare, Simon; Hale, Christine; Seshasayee, Aswin; Harris, Simon; Thomson, Nicholas R; Gardner, Paul; Rabsch, Wolfgang; Wigley, Paul; Humphrey, Tom; Parkhill, Julian; Dougan, Gordon

2013-08-27

Salmonella enterica serovar Typhimurium definitive type 2 (DT2) is host restricted to Columba livia (rock or feral pigeon) but is also closely related to S. Typhimurium isolates that circulate in livestock and cause a zoonosis characterized by gastroenteritis in humans. DT2 isolates formed a distinct phylogenetic cluster within S. Typhimurium based on whole-genome-sequence polymorphisms. Comparative genome analysis of DT2 94-213 and S. Typhimurium SL1344, DT104, and D23580 identified few differences in gene content with the exception of variations within prophages. However, DT2 94-213 harbored 22 pseudogenes that were intact in other closely related S. Typhimurium strains. We report a novel in silico approach to identify single amino acid substitutions in proteins that have a high probability of a functional impact. One polymorphism identified using this method, a single-residue deletion in the Tar protein, abrogated chemotaxis to aspartate in vitro. DT2 94-213 also exhibited an altered transcriptional profile in response to culture at 42°C compared to that of SL1344. Such differentially regulated genes included a number involved in flagellum biosynthesis and motility. IMPORTANCE Whereas Salmonella enterica serovar Typhimurium can infect a wide range of animal species, some variants within this serovar exhibit a more limited host range and altered disease potential. Phylogenetic analysis based on whole-genome sequences can identify lineages associated with specific virulence traits, including host adaptation. This study represents one of the first to link pathogen-specific genetic signatures, including coding capacity, genome degradation, and transcriptional responses to host adaptation within a Salmonella serovar. We performed comparative genome analysis of reference and pigeon-adapted definitive type 2 (DT2) S. Typhimurium isolates alongside phenotypic and transcriptome analyses, to identify genetic signatures linked to host adaptation within the DT2 lineage.

PLASMID PROFILES AND PHAGE TYPES OF SALMONELLA-TYPHIMURIUM ISOLATED FROM SUCCESSIVE FLOCKS OF CHICKENS ON 3 PARENT STOCK FARMS

DEFF Research Database (Denmark)

Baggesen, Dorte Lau; Olsen, J. E.; Bisgaard, M.

1992-01-01

Three-hundred-and-eighty-seven strains of Salmonella typhimurium obtained from successive generations of parent stock originating from three different rearing farms were characterized by phage typing and plasmid profiling. Seventy-six strains representing dominant types were selected for restrict......Three-hundred-and-eighty-seven strains of Salmonella typhimurium obtained from successive generations of parent stock originating from three different rearing farms were characterized by phage typing and plasmid profiling. Seventy-six strains representing dominant types were selected...

Colicinogeny in Salmonella serovars isolated in Brazil

Directory of Open Access Journals (Sweden)

Leila Carvalho Campos

1988-06-01

Full Text Available A study of colicinogeny was made in 748 strains of Salmonella (97 serovars isolated from different sources; human (291, animal (119, environmental (141, food (102 and animal feed (95. Colicin production was detected in 64 strains (8.6%, particularly isolated from foods (30.4%. Col. E1 (53 and Ia (44 were the most frequently observed, especially in S. agona for environment and food sources. Col V production was identified in 5 strains of S. typhimurium within 8 producer cultures isolated from humans. Its relationship with the sources and serovars of Salmonella are discussed.Investigou-se a produção de colicina em 748 amostras de Salmonella (97 sorovares advindas de díferentes fontes: humana (291, animal (119, ambiental (141, de alimentos (102 e rações (95. Detectaram-se 64 amostras (8,6% colicinogênicas, particularmente isoladas de alimentos (30,4%. ColE1 (53 e Ia (44 foram as mais freqüentes, especialmente no sorovar S, agona, de origem ambiental e de alimentos. Identificou-se também a produção de col V em 5 amostras de S. typhimurium dentre 8 culturas produtoras de origem humana. Discute-se a relação entre a capacidade colicinogênica e as fontes e sorovares de Salmonella.

Characterization of integron mediated antimicrobial resistance in Salmonella isolated from diseased swine

Science.gov (United States)

White, David G.; Zhao, Shaohua; McDermott, Patrick F.; Ayers, Sherry; Friedman, Sharon; Sherwood, Julie; Breider-Foley, Missy; Nolan, Lisa K.

2003-01-01

Forty-two Salmonella isolates obtained from diseased swine were genetically characterized for the presence of specific antimicrobial resistance mechanisms. Twenty of these isolates were characterized as S. Typhimurium DT104 strains. Pulsed-field gel electrophoresis was used to determine genetic relatedness and revealed 20 distinct genetic patterns among the 42 isolates. However, all DT104 isolates fell within 2 closely related genetic clusters. Other Salmonella isolates were genetically grouped together according to serotype. All DT104 isolates displayed the penta-resistance phenotype to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline. Resistance to sulfamethoxazole, tetracycline, streptomycin, kanamycin, and ampicillin was most common among the non-DT104 Salmonella isolates. All DT104 strains contained 2 chromosomal integrons of 1000 and 1200 base pairs. The DNA sequencing revealed that the 2 integrons contained genes encoding a resistance to streptomycin and ampicillin, respectively. None of the non-DT104 strains showed the same pattern, although several strains possessed integrons of 1000 base pairs or larger. However, the majority of non-DT104 Salmonella strains did not possess any integrons. Two Salmonella isolates displayed tolerance to the organic solvent cyclohexane, indicating the possibility that they are overexpressing chromosomal regulatory genes marA or soxS or the associated multidrug efflux pump, acrAB. This research suggests that integrons contribute to antimicrobial resistance among specific swine Salmonella serotypes; however, they are not as widely disseminated among non-Typhimurium swine Salmonella serotypes as previously thought. PMID:12528827

Survival and transmission of Salmonella enterica serovar typhimurium in an outdoor organic pig farming environment

DEFF Research Database (Denmark)

Jensen, Annette Nygaard; Dalsgaard, Anders; Stockmarr, Anders

2006-01-01

It was investigated how organic rearing conditions influence the Salmonella enterica infection dynamics in pigs and whether Salmonella persists in the paddock environment. Pigs inoculated with S. enterica serovar Typhimurium were grouped with Salmonella-negative tracer pigs. Bacteriological...... the seroprevalence. Salmonella persisted in the paddock environment, as Salmonella was isolated from 46% of soil and water samples (n = 294). After removal of pigs, Salmonella was found in soil samples for up to. 5 weeks and in shelter huts during the entire test period (7 weeks). Subsequent introduction...... of Salmonella-negative pigs into four naturally Salmonella-contaminated paddocks caused Salmonella infections of pigs in two paddocks. In one of these paddocks, all tracer pigs (n = 10) became infected, coinciding with a previous high Salmonella infection rate and high Salmonella excretion level. Our results...

Salmonella typhimurium DT104: a virulent and drug-resistant pathogen.

OpenAIRE

Poppe, C; Smart, N; Khakhria, R; Johnson, W; Spika, J; Prescott, J

1998-01-01

Salmonella typhimurium phage type (PT) or definitive type (DT) 104 is a virulent pathogen for humans and animals, particularly cattle. It has been isolated increasingly from humans and animals in the United Kingdom and several other European countries and, more recently, in the United States and Canada. Humans may acquire the infection from foods of animal origin contaminated with the infective organism. Farm families are particularly at risk of acquiring the infection by contact with infecte...

Arginine-dependent acid resistance in Salmonella enterica serovar Typhimurium

NARCIS (Netherlands)

Kieboom, J.; Abee, T.

2006-01-01

Salmonella enterica serovar Typhimurium does not survive a pH 2.5 acid challenge under conditions similar to those used for Escherichia coli (J. W. Foster, Nat. Rev. Microbiol. 2:898-907, 2004). Here, we provide evidence that S. enterica serovar Typhimurium can display arginine-dependent acid

Molecular Characterization and Antimicrobial Susceptibility of Salmonella Isolates from Infections in Humans in Henan Province, China

DEFF Research Database (Denmark)

Xia, S.L.; Hendriksen, Rene S.; Xie, Z.Q.

2009-01-01

We characterized 208 human Salmonella isolates from 2006 to 2007 and 27 human Salmonella enterica serovar Typhimurium isolates from 1987 to 1993 from Henan Province, China, by serotyping, by antimicrobial susceptibility testing, and, for the most common serovars, by pulsed-field gel electrophoresis...... (PFGE). The most common serovars among the 2006-2007 isolates were S. enterica serovar Typhimurium (27%), S. enterica serovar Enteritidis (17%), S. enterica serovar Derby (10%), S. enterica serovar Indiana (6%), and S. enterica serovar Litchfield (6%). A high percentage of the isolates were multiple-drug...

Resistance of Salmonella enteritidis variety typhimurium to gamma radiation

International Nuclear Information System (INIS)

Norberg, A.N.; Maliska, C.

1988-01-01

The use of ionizing radiations to kill microrganisms responsible for food deterioration, and toxinfections is an example of peaceful use of nuclear energy. Food toxinfections are, amongus, produced mostly by Salmonella enteritidis var. typhimurium. Due to the pauncity of information on the resistance to gamma radiation of Salmonella enteritidis var. typhimurium this paper has the aim to define the 60-Cobalt gamma radiation lethal dose to these bacteria, in experimentally contaminated milk by samples recovered from our geographycal area. One hundred nineteen samples of milk containing about 150.000 bacteria per ml were irradiated with doses ranging from 100 to 1.100 Gy. Two samples of surving bacteria were again irradiated by doses up to 2.500 Gy. The bacteria not previously irradiated were killed by doses of 1.100 Gy. It was concluded that the 60-Cobalt gamma radiation minimal lethal dose to Salmonella enteritidis var. typhimurium is 1.200 Gy. The surviving strains to smaller doses than 1.200 Gy when re-irradiated prompt the forthcoming of more radio-resistant germs. (author) [pt

Pathogenicity of Salmonella Strains Isolated from Egg Shells and the Layer Farm Environment in Australia

Science.gov (United States)

McWhorter, Andrea R.; Davos, Dianne

2014-01-01

In Australia, the egg industry is periodically implicated during outbreaks of Salmonella food poisoning. Salmonella enterica serovar Typhimurium and other nontyphoidal Salmonella spp., in particular, are a major concern for Australian public health. Several definitive types of Salmonella Typhimurium strains, but primarily Salmonella Typhimurium definitive type 9 (DT9), have been frequently reported during egg-related food poisoning outbreaks in Australia. The aim of the present study was to generate a pathogenicity profile of nontyphoidal Salmonella isolates obtained from Australian egg farms. To achieve this, we assessed the capacity of Salmonella isolates to cause gastrointestinal disease using both in vitro and in vivo model systems. Data from in vitro experiments demonstrated that the invasion capacity of Salmonella serovars cultured to stationary phase (liquid phase) in LB medium was between 90- and 300-fold higher than bacterial suspensions in normal saline (cultured in solid phase). During the in vivo infection trial, clinical signs of infection and mortality were observed only for mice infected with either 103 or 105 CFU of S. Typhimurium DT9. No mortality was observed for mice infected with Salmonella serovars with medium or low invasive capacity in Caco-2 cells. Pathogenicity gene profiles were also generated for all serovars included in this study. The majority of serovars tested were positive for selected virulence genes. No relationship between the presence or absence of virulence genes by PCR and either in vitro invasive capacity or in vivo pathogenicity was detected. Our data expand the knowledge of strain-to-strain variation in the pathogenicity of Australian egg industry-related Salmonella spp. PMID:25362057

Comparative genome analysis of the high pathogenicity Salmonella Typhimurium strain UK-1.

Directory of Open Access Journals (Sweden)

Yingqin Luo

Full Text Available Salmonella enterica serovar Typhimurium, a gram-negative facultative rod-shaped bacterium causing salmonellosis and foodborne disease, is one of the most common isolated Salmonella serovars in both developed and developing nations. Several S. Typhimurium genomes have been completed and many more genome-sequencing projects are underway. Comparative genome analysis of the multiple strains leads to a better understanding of the evolution of S. Typhimurium and its pathogenesis. S. Typhimurium strain UK-1 (belongs to phage type 1 is highly virulent when orally administered to mice and chickens and efficiently colonizes lymphoid tissues of these species. These characteristics make this strain a good choice for use in vaccine development. In fact, UK-1 has been used as the parent strain for a number of nonrecombinant and recombinant vaccine strains, including several commercial vaccines for poultry. In this study, we conducted a thorough comparative genome analysis of the UK-1 strain with other S. Typhimurium strains and examined the phenotypic impact of several genomic differences. Whole genomic comparison highlights an extremely close relationship between the UK-1 strain and other S. Typhimurium strains; however, many interesting genetic and genomic variations specific to UK-1 were explored. In particular, the deletion of a UK-1-specific gene that is highly similar to the gene encoding the T3SS effector protein NleC exhibited a significant decrease in oral virulence in BALB/c mice. The complete genetic complements in UK-1, especially those elements that contribute to virulence or aid in determining the diversity within bacterial species, provide key information in evaluating the functional characterization of important genetic determinants and for development of vaccines.

Virulence-associated genes, antimicrobial resistance and molecular typing of Salmonella Typhimurium strains isolated from swine from 2000 to 2012 in Brazil.

Science.gov (United States)

Almeida, F; Medeiros, M I C; Kich, J D; Falcão, J P

2016-06-01

The aims of this study were to assess the pathogenic potential, antimicrobial resistance and genotypic diversity of Salmonella Typhimurium strains isolated in Brazil from swine (22) and the surrounding swine environment (5) from 2000 to 2012 and compare them to the profiles of 43 human strains isolated from 1983 to 2010, which had been previously studied. The presence of 12 SPI-1, SPI-2 and plasmid genes was assessed by PCR, the antimicrobial susceptibility to 13 antimicrobials was determined by the disc diffusion assay and genotyping was performed using pulsed-field gel electrophoresis (PFGE), multiple-locus variable-number of tandem repeats analysis (MLVA) and ERIC-PCR. More than 77·8% of the swine strains carried 10 or more of the virulence markers. Ten (37%) strains isolated from swine were multi-drug resistant (MDR). All the molecular typing techniques grouped the strains in two main clusters. Some strains isolated from swine and humans were allocated together in the PFGE-B2, MLVA-A1, MLVA-B and ERIC-A1 clusters. The genotyping results suggest that some strains isolated from swine and humans may descend from a common subtype and may indicate a possible risk of MDR S. Typhimurium with high frequency of virulence genes isolated from swine to contaminate humans in Brazil. This study provided new information about the pathogenic potential, antimicrobial resistance and genotypic diversity of S. Typhimurium isolates from swine origin in Brazil, the fourth largest producer of pigs worldwide. © 2016 The Society for Applied Microbiology.

Tetracycline consumption and occurrence of tetracycline resistance in Salmonella typhimurium phage types from Danish pigs

DEFF Research Database (Denmark)

Emborg, Hanne-Dorthe; Vigre, Håkan; Jensen, Vibeke Frøkjær

2007-01-01

more than doubled at the national level from 12,000-13,000 kg of active compound in 1996-1998 to 29,000 kg of active compound in 2004. Instead, tetracycline-resistant S. Typhimurium phage types became more prevalent. This suggests that the spread of already established or new resistant clones, rather......The aims of the present study were to investigate at the farm-owner level the effect of prescribed tetracycline consumption in pigs and different Salmonella Typhimurium phage types on the probability that the S. Typhimurium was resistant to tetracycline. In this study, 1,307 isolates were included......, originating from 877 farm owners, and data were analyzed using logistic regression. The analysis showed that both the S. Typhimurium phage type (p type...

The detection of Salmonella typhimurium on shell eggs using a phage-based biosensor

Science.gov (United States)

Chai, Yating; Li, Suiqiong; Horikawa, Shin; Shen, Wen; Park, Mi-Kyung; Vodyanoy, Vitaly J.; Chin, Bryan A.

2011-06-01

This paper presents the direct detection of Salmonella typhimurium on shell eggs using a phage-based magnetoelastic (ME) biosensor. The ME biosensor consists of a ME resonator as the sensor platform and E2 phage as the biorecognition element that is genetically engineered to specifically bind with Salmonella typhimurium. The ME biosensor, which is a wireless sensor, vibrates with a characteristic resonant frequency under an externally applied magnetic field. Multiple sensors can easily be remotely monitored. Multiple measurement and control sensors were placed on the shell eggs contaminated by Salmonella typhimurium solutions with different known concentrations. The resonant frequency of sensors before and after the exposure to the spiked shell eggs was measured. The frequency shift of the measurement sensors was significantly different than the control sensors indicating Salmonella contamination. Scanning electron microscopy was used to confirm binding of Salmonella to the sensor surface and the resulting frequency shift results.

Fate of Salmonella Typhimurium in laboratory-scale drinking water biofilms

CSIR Research Space (South Africa)

Schaefer, Lisa M

2013-08-01

Full Text Available biofilms in monoculture and the fate and persistence of Salmonella in a mixed aquatic biofilm was examined. In monoculture S. Typhimurium formed loosely structured biofilms. Salmonella colonized established multi-species drinking water biofilms within 24...

Interaction between Salmonella typhimurium and phagocytic cells in pigs - Phagocytosis, oxidative burst and killing in polymorphonuclear leukocytes and monocytes

DEFF Research Database (Denmark)

Riber, Ulla; Lind, Peter

1999-01-01

Interactions between Salmonella typhimurium and peripheral blood leucocytes from healthy, Salmonella-free pigs were investigated in vitro. Both granulocytes and monocytes phagocytized FITC-labelled heat-killed Salmonella bacteria as shown by flow cytometry. Phagocytosis in whole blood and isolated...... with the exhaustion of oxidative burst in non-adherent monocytes were performed by prestimulation with PMA, heat-killed Salmonella or buffer. Prestimulation with PMA led to a strong reduction in oxidative burst induced by living opsonized Salmonella bacteria, whereas prestimulation with heat-killed bacteria gave rise...

Virulence Characterization of Salmonella enterica by a New Microarray: Detection and Evaluation of the Cytolethal Distending Toxin Gene Activity in the Unusual Host S. Typhimurium.

Directory of Open Access Journals (Sweden)

Rui Figueiredo

Full Text Available Salmonella enterica is a zoonotic foodborne pathogen that causes acute gastroenteritis in humans. We assessed the virulence potential of one-hundred and six Salmonella strains isolated from food animals and products. A high through-put virulence genes microarray demonstrated Salmonella Pathogenicity Islands (SPI and adherence genes were highly conserved, while prophages and virulence plasmid genes were variably present. Isolates were grouped by serotype, and virulence plasmids separated S. Typhimurium in two clusters. Atypical microarray results lead to whole genome sequencing (WGS of S. Infantis Sal147, which identified deletion of thirty-eight SPI-1 genes. Sal147 was unable to invade HeLa cells and showed reduced mortality in Galleria mellonella infection model, in comparison to a SPI-1 harbouring S. Infantis. Microarray and WGS of S. Typhimurium Sal199, established for the first time in S. Typhimurium presence of cdtB and other Typhi-related genes. Characterization of Sal199 showed cdtB genes were upstream of transposase IS911, and co-expressed with other Typhi-related genes. Cell cycle arrest, cytoplasmic distension, and nuclear enlargement were detected in HeLa cells infected by Sal199, but not with S. Typhimurium LT2. Increased mortality of Galleria was detected on infection with Sal199 compared to LT2. Thus, Salmonella isolates were rapidly characterized using a high through-put microarray; helping to identify unusual virulence features which were corroborated by further characterisation. This work demonstrates that the use of suitable screening methods for Salmonella virulence can help assess the potential risk associated with certain Salmonella to humans. Incorporation of such methodology into surveillance could help reduce the risk of emergence of epidemic Salmonella strains.

Antibiotic resistant Salmonella and Escherichia coli isolated from ...

African Journals Online (AJOL)

Results: A hundred and four indigenous chicken rectal swabs were analysed, of which 67.3% were contaminated with Escherichia coli and 12.5% with Salmonella typhimurium. Seventy Escherichia coli isolates showed resistance phenotypes to one, two or more antibiotics. The most common antimicrobial resistance pattern ...

β-1,3/1,6-Glucan alleviated intestinal mucosal barrier impairment of broiler chickens challenged with Salmonella enterica serovar Typhimurium.

Science.gov (United States)

Shao, Yujing; Guo, Yuming; Wang, Zhong

2013-07-01

This study investigated the protective effect of β-1,3/1,6-glucan on gut morphology, intestinal epithelial tight junctions, and bacterial translocation of broiler chickens challenged with Salmonella enterica serovar Typhimurium. Ninety Salmonella-free Arbor Acre male broiler chickens were randomly divided into 3 groups: negative control group (NC), Salmonella Typhimurium-infected positive group (PC), and the Salmonella Typhimurium-infected group with dietary 100 mg/kg of β-1,3/1,6-glucan supplementation (T) to determine the effect of β-1,3/1,6-glucan on intestinal barrier function. Salmonella Typhimurium challenge alone significantly decreased villus height (P chickens challenged with Salmonella Typhimurium.

Study of Salmonella typhimurium mutagenicity assay of (E ...

African Journals Online (AJOL)

Study of Salmonella typhimurium mutagenicity assay of (E)-piplartine by the Ames test. AA Morandim-Giannetti, F Cotinguiba, LO Regasini, MC Frigieri, EA Varanda, A Coqueiro, MJ Kato, VS Bolzani, M Furlan ...

The inhibitory effect of bovine rumen fluid on Salmonella typhimurium.

Science.gov (United States)

Chambers, P G; Lysons, R J

1979-05-01

The possible fate of Salmonella typhimurium in the rumen was investigated by monitoring rumen volatile fatty acids (VFA), lactate concentrations and pH over periods which included regular feeding and 48 h starvation. Preparations were made containing 50 per cent rumen fluid from the cow or VFA solutions, and then inoculated with S typhimurium. Viable counts before and after incubation for 24 h at 37 degrees C were compared. Incubation in broths with high concentrations of VFA and low pH resulted in a marked decrease in salmonella numbers, while lower VFA concentrations had little or no inhibitory effect on growth.

Use of multiple-locus variable-number tandem-repeats analysis (MLVA) typing to characterize Salmonella Typhimurium DT41 broiler breeder infections

DEFF Research Database (Denmark)

Litrup, E.; Christensen, H.; Nordentoft, Steen

2010-01-01

To characterize isolates of Salmonella Typhimurium DT41 obtained from infected flocks of broiler breeders by multiple-locus variable-number tandem-repeats analysis (MLVA) and compare results with a diverse strain collection from Germany and United Kingdom and isolates from Danish patients. A total...

Molecular Confirmation of Salmonella typhimuriumin Poultry from Kathmandu Valley

Directory of Open Access Journals (Sweden)

Sanjeev Kumar Adhikari

2018-05-01

Full Text Available A prevalence study was carried to isolate Salmonella typhimurium from blood (n= 50 and gut samples (n=100 of poultry in Kathmandu valley during early 2016. Salmonella typhimurium bacteria isolated in the selective media were biochemically confirmed based on Bergey’s Manual. Two sets of oligonucleotide primers-the genus specific 16S rRNA and the organism specific invA were employed for molecular level confirmation by the Polymerase Chain Reaction (PCR assay. The amplified fragments in 1% agarose gel observed at 406bp and 285bp, respectively confirmed the isolates to be Salmonella typhimurium. Of 150 samples tested, Salmonella typhimurium were isolated from 49 samples, among which nine were from blood (18% and forty from the gut (40%. The present result indicated an alarmingly high level of Salmonella typhimurium, which can result inzoonotic infection in humans owing to increased contact with poultry and consumption of poultry products in the Kathmandu valley.

Biological effect of plutonium 239 on Salmonella typhimurium

International Nuclear Information System (INIS)

Gafieva, Z.A.; Chudin, V.A.

1988-01-01

Salmonella typhimurium cells were exposed in a 239 Pu citrate solution. Cell death and induction of gene mutations were an exponential fucntion of γ-radiation dose. LD 37 was 34.8 Gy; mutation doubling dose, 19 Gy

Detoxification of Salmonella typhimurium lipopolysaccharide by ionizing radiation.

Science.gov (United States)

Previte, J J; Chang, Y; el-Bisi, H M

1967-05-01

The efficiency of ionizing radiation in detoxifying the lethal determinant(s) of the lipopolysaccharide (LPS) of Salmonella typhimurium, S. enteritidis, and Escherichia coli in aqueous solution and associated with heat-killed S. typhimurium cells in suspension decreased with doses above 1 Mrad. The 50% end point of inactivation was more than 7.0 Mrad for heat-killed salmonellae and 4.8, 4.5, and 1.0 Mrad for the LPS of S. typhimurium, S. enteritidis, and E. coli, respectively. After exposure to 20 Mrad, S. typhimurium LPS retained a small portion of its lethal properties although the ld(50) was much greater than 9.5 mg per 20-g mouse. However, at -184 C, no inactivation of the lethal determinant(s) occurred after exposure to as much as 20 Mrad. This demonstrated the significance of the indirect effect and the mobility and formation of free radicals. At 22 C, the optical density at 400 mmu increased and the pH decreased with increasing radiation dose, but no qualitative changes were observed in the infrared spectrum. No change was observed in the pyrogenicity of S. typhimurium LPS; a slight decrease in antigenicity was revealed when 6 days, but not when 1 day, elapsed between vaccination and challenge in the mouse protection test. The results were interpreted as evidence of the existence of two or more lethal and antigenic determinants. The differential effect of radiation on these properties and on the pyrogenic component(s) probably are indicative of separate functional sites for lethal, antigenic, and pyrogenic activities.

Immunochromatographic strip assay for the rapid and sensitive detection of Salmonella Typhimurium in artificially contaminated tomato samples.

Science.gov (United States)

Shukla, Shruti; Leem, Hyerim; Lee, Jong-Suk; Kim, Myunghee

2014-06-01

This study was designed to confirm the applicability of a liposome-based immunochromatographic assay for the rapid detection of Salmonella enterica subsp. enterica serovar Typhimurium (Salmonella Typhimurium) in artificially contaminated tomato samples. To determine the detection limit and pre-enrichment incubation time (10, 12, and 18 h pre-enrichment in 1% buffered peptone water), the tests were performed with different cell numbers of Salmonella Typhimurium (3 × 10(0), 3 × 10(1), 3 × 10(2), and 3 × 10(3) CFU·mL(-1)) inoculated into 25 g of crushed tomato samples. The assay was able to detect as few as 30 Salmonella Typhimurium cells per 25 g of tomato samples (1.2 cells·g(-1)) after 12 h pre-enrichment incubation. Moreover, when the developed assay was compared with traditional morphological and biochemical culture-based methods as well as colloidal gold nanoparticle-based commercial test strips, the developed assay yielded positive results for the detection of Salmonella Typhimurium within a shorter period time. These findings confirm that the developed assay may have practical application for the sensitive detection of Salmonella Typhimurium in various food samples, including raw vegetables, with a relatively low detection limit and shorter analysis time.

Investigation and management of an outbreak of Salmonella Typhimurium DT8 associated with duck eggs, Ireland 2009 to 2011.

LENUS (Irish Health Repository)

Garvey, P

2013-01-01

Salmonella Typhimurium DT8 was a very rare cause of human illness in Ireland between 2000 and 2008, with only four human isolates from three patients being identified. Over a 19-month period between August 2009 and February 2011, 34 confirmed cases and one probable case of Salmonella Typhimurium DT8 were detected, all of which had an MLVA pattern 2-10-NA-12-212 or a closely related pattern. The epidemiological investigations strongly supported a linkbetween illness and exposure to duck eggs. Moreover, S. Typhimurium with an MLVA pattern indistinguishable (or closely related) to the isolates from human cases, was identified in 22 commercial and backyard duck flocks, twelve of which were linked with known human cases. A range of control measures were taken at farm level, and advice was provided to consumers on the hygienic handling and cooking of duck eggs. Although no definitive link was established with a concurrent duck egg-related outbreak of S. Typhimurium DT8 in the United Kingdom, it seems likely that the two events were related. It may be appropriate for other countries with a tradition of consuming duck eggs to consider the need for measures to reduce the risk of similar outbreaks.

Isolation, crystallization and preliminary crystallographic analysis of Salmonella typhimurium uridine phosphorylase crystallized with 2,2′-anhydrouridine

Energy Technology Data Exchange (ETDEWEB)

Timofeev, Vladimir I.; Lashkov, Alexander A.; Gabdoulkhakov, Azat G.; Pavlyuk, Bogdan Ph. [A. V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninskiy Prospect 59, 119333 Moscow (Russian Federation); Kachalova, Galina S. [Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Institutskaya Street 3, 142290 Pushchino, Moscow Region (Russian Federation); Betzel, Christian [Institute fur Biochemie und Lebensmittelchemie, University of Hamburg, c/o DESY, Building 22, Notkestrasse 85, 22604 Hamburg (Germany); Morgunova, Ekaterina Yu.; Zhukhlistova, Nadezhda E.; Mikhailov, Al’bert M., E-mail: amm@ns.crys.ras.ru [A. V. Shubnikov Institute of Crystallography, Russian Academy of Sciences, Leninskiy Prospect 59, 119333 Moscow (Russian Federation)

2007-10-01

S. typhimurium uridine phosphorylase has been isolated and crystallized in the presence of ligand. Uridine phosphorylase (UPh; EC 2.4.2.3) is a member of the pyrimidine nucleoside phosphorylase family of enzymes which catalyzes the phosphorolytic cleavage of the C—N glycoside bond of uridine, with the formation of ribose 1-phosphate and uracil. This enzyme has been shown to be important in the activation and catabolism of fluoropyrimidines. Modulation of its enzymatic activity may affect the therapeutic efficacy of chemotherapeutic agents. The structural investigation of the bacterial uridine phosphorylases, both unliganded and complexed with substrate/product analogues and inhibitors, may help in understanding the catalytic mechanism of the phosphorolytic cleavage of uridine. Salmonella typhimurium uridine phosphorylase has been crystallized with 2,2′-anhydrouridine. X-ray diffraction data were collected to 2.15 Å. Preliminary analysis of the diffraction data indicates that the crystal belongs to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 88.52, b = 123.98, c = 133.52 Å. The solvent content is 45.51%, assuming the presence of one hexamer molecule per asymmetric unit.

Salmonella enterica serovar Typhimurium exploits inflammation to modify swine intestinal microbiota.

Directory of Open Access Journals (Sweden)

Rosanna eDrumo

2016-01-01

Full Text Available Salmonella enterica serovar Typhimurium is an important zoonotic gastrointestinal pathogen responsible for foodborne disease worldwide. It is a successful enteric pathogen because it has developed virulence strategies allowing it to survive in a highly inflamed intestinal environment exploiting inflammation to overcome colonization resistance provided by intestinal microbiota. In this study, we used piglets featuring an intact microbiota, which naturally develop gastroenteritis, as model for salmonellosis. We compared the effects on the intestinal microbiota induced by a wild type and an attenuated S. Typhimurium in order to evaluate whether the modifications are correlated with the virulence of the strain. This study showed that Salmonella alters microbiota in a virulence-dependent manner. We found that the wild type S. Typhimurium induced inflammation and a reduction of specific protecting microbiota species (SCFA-producing bacteria normally involved in providing a barrier against pathogens. Both these effects could contribute to impair colonization resistance, increasing the host susceptibility to wild type S. Typhimurium colonization. In contrast, the attenuated S. Typhimurium, which is characterized by a reduced ability to colonize the intestine, and by a very mild inflammatory response, was unable to successfully sustain competition with the microbiota.

EVALUATION OF AN O-ANTIGEN ELISA FOR SCREENING CATTLE HERDS FOR SALMONELLA-TYPHIMURIUM

DEFF Research Database (Denmark)

Hoorfar, Jeffrey; Bitsch, V.

1995-01-01

A total of 2585 serum samples from 62 dairy herds located in four different regions of Denmark were tested in an O-antigen (0:1,4,5,12)-based ELISA for the detection of antibodies against Salmonella typhimurium. Ten closed herds from an island with no reported occurrence of salmonellosis for seve......A total of 2585 serum samples from 62 dairy herds located in four different regions of Denmark were tested in an O-antigen (0:1,4,5,12)-based ELISA for the detection of antibodies against Salmonella typhimurium. Ten closed herds from an island with no reported occurrence of salmonellosis...... for several years, and 12 herds from a salmonella enzootic area which had had clinical outbreaks of S typhimurium were used to define a herd ELISA cut-off value. When herds with at least 5 per cent of the serum samples having an optical density of >0.5 were considered ELISA-positive, all 10 herds from...... the salmonellosis-free island were ELISA-negative, and all but one of the 12 S typhimurium-infected herds were ELISA-positive, which resulted in a herd test sensitivity of 0.92 and herd test specificity of 1.0. Eleven of the 12 S typhimurium-infected herds were negative in a blocking ELISA based on a monoclonal...

Virulence of invasive Salmonella Typhimurium ST313 in animal models of infection.

Directory of Open Access Journals (Sweden)

Girish Ramachandran

2017-08-01

Full Text Available Salmonella Typhimurium sequence type (ST 313 produces septicemia in infants in sub-Saharan Africa. Although there are known genetic and phenotypic differences between ST313 strains and gastroenteritis-associated ST19 strains, conflicting data about the in vivo virulence of ST313 strains have been reported. To resolve these differences, we tested clinical Salmonella Typhimurium ST313 and ST19 strains in murine and rhesus macaque infection models. The 50% lethal dose (LD50 was determined for three Salmonella Typhimurium ST19 and ST313 strains in mice. For dissemination studies, bacterial burden in organs was determined at various time-points post-challenge. Indian rhesus macaques were infected with one ST19 and one ST313 strain. Animals were monitored for clinical signs and bacterial burden and pathology were determined. The LD50 values for ST19 and ST313 infected mice were not significantly different. However, ST313-infected BALB/c mice had significantly higher bacterial numbers in blood at 24 h than ST19-infected mice. ST19-infected rhesus macaques exhibited moderate-to-severe diarrhea while ST313-infected monkeys showed no-to-mild diarrhea. ST19-infected monkeys had higher bacterial burden and increased inflammation in tissues. Our data suggest that Salmonella Typhimurium ST313 invasiveness may be investigated using mice. The non-human primate results are consistent with clinical data, suggesting that ST313 strains do not cause diarrhea.

Virulence of invasive Salmonella Typhimurium ST313 in animal models of infection.

Science.gov (United States)

Ramachandran, Girish; Panda, Aruna; Higginson, Ellen E; Ateh, Eugene; Lipsky, Michael M; Sen, Sunil; Matson, Courtney A; Permala-Booth, Jasnehta; DeTolla, Louis J; Tennant, Sharon M

2017-08-01

Salmonella Typhimurium sequence type (ST) 313 produces septicemia in infants in sub-Saharan Africa. Although there are known genetic and phenotypic differences between ST313 strains and gastroenteritis-associated ST19 strains, conflicting data about the in vivo virulence of ST313 strains have been reported. To resolve these differences, we tested clinical Salmonella Typhimurium ST313 and ST19 strains in murine and rhesus macaque infection models. The 50% lethal dose (LD50) was determined for three Salmonella Typhimurium ST19 and ST313 strains in mice. For dissemination studies, bacterial burden in organs was determined at various time-points post-challenge. Indian rhesus macaques were infected with one ST19 and one ST313 strain. Animals were monitored for clinical signs and bacterial burden and pathology were determined. The LD50 values for ST19 and ST313 infected mice were not significantly different. However, ST313-infected BALB/c mice had significantly higher bacterial numbers in blood at 24 h than ST19-infected mice. ST19-infected rhesus macaques exhibited moderate-to-severe diarrhea while ST313-infected monkeys showed no-to-mild diarrhea. ST19-infected monkeys had higher bacterial burden and increased inflammation in tissues. Our data suggest that Salmonella Typhimurium ST313 invasiveness may be investigated using mice. The non-human primate results are consistent with clinical data, suggesting that ST313 strains do not cause diarrhea.

Comparison between digital PCR and real-time PCR in detection of Salmonella typhimurium in milk.

Science.gov (United States)

Wang, Meng; Yang, Junjie; Gai, Zhongtao; Huo, Shengnan; Zhu, Jianhua; Li, Jun; Wang, Ranran; Xing, Sheng; Shi, Guosheng; Shi, Feng; Zhang, Lei

2018-02-02

As a kind of zero-tolerance foodborne pathogens, Salmonella typhimurium poses a great threat to quality of food products and public health. Hence, rapid and efficient approaches to identify Salmonella typhimurium are urgently needed. Combined with PCR and fluorescence technique, real-time PCR (qPCR) and digital PCR (ddPCR) are regarded as suitable tools for detecting foodborne pathogens. To compare the effect between qPCR and ddPCR in detecting Salmonella typhimurium, a series of nucleic acid, pure strain culture and spiking milk samples were applied and the resistance to inhibitors referred in this article as well. Compared with qPCR, ddPCR exhibited more sensitive (10 -4 ng/μl or 10 2 cfu/ml) and less pre-culturing time (saving 2h). Moreover, ddPCR had stronger resistance to inhibitors than qPCR, yet absolute quantification hardly performed when target's concentration over 1ng/μl or 10 6 cfu/ml. This study provides an alternative strategy in detecting foodborne Salmonella typhimurium. Copyright © 2018 Elsevier B.V. All rights reserved.

Epidemiology of a Salmonella enterica subsp. Enterica serovar Typhimurium strain associated with a songbird outbreak.

Science.gov (United States)

Blehert, David S.; Hernandez, Sonia M.; Keel, Kevin; Sanchez, Susan; Trees, Eija; ,

2012-01-01

Salmonella enterica subsp. enterica serovar Typhimurium is responsible for the majority of salmonellosis cases worldwide. This Salmonella serovar is also responsible for die-offs in songbird populations. In 2009, there was an S. Typhimurium epizootic reported in pine siskins in the eastern United States. At the time, there was also a human outbreak with this serovar that was associated with contaminated peanuts. As peanuts are also used in wild-bird food, it was hypothesized that the pine siskin epizootic was related to this human outbreak. A comparison of songbird and human S. Typhimurium pulsed-field gel electrophoresis (PFGE) patterns revealed that the epizootic was attributed not to the peanut-associated strain but, rather, to a songbird strain first characterized from an American goldfinch in 1998. This same S. Typhimurium strain (PFGE type A3) was also identified in the PulseNet USA database, accounting for 137 of 77,941 total S. Typhimurium PFGE entries. A second molecular typing method, multiple-locus variable-number tandem-repeat analysis (MLVA), confirmed that the same strain was responsible for the pine siskin epizootic in the eastern United States but was distinct from a genetically related strain isolated from pine siskins in Minnesota. The pine siskin A3 strain was first encountered in May 2008 in an American goldfinch and later in a northern cardinal at the start of the pine siskin epizootic. MLVA also confirmed the clonal nature of S. Typhimurium in songbirds and established that the pine siskin epizootic strain was unique to the finch family. For 2009, the distribution of PFGE type A3 in passerines and humans mirrored the highest population density of pine siskins for the East Coast.

The transcriptional landscape and small RNAs of Salmonella enterica serovar Typhimurium

DEFF Research Database (Denmark)

Kröger, Carsten; Dillon, Shane C.; Cameron, Andrew D. S.

2012-01-01

More than 50 y of research have provided great insight into the physiology, metabolism, and molecular biology of Salmonella enterica serovar Typhimurium (S. Typhimurium), but important gaps in our knowledge remain. It is clear that a precise choreography of gene expression is required......-thirds of these TSSs were associated with σ70 (including phoP, slyA, and invF) from which we identified the −10 and −35 motifs of σ70-dependent S. Typhimurium gene promoters. Overall, we corrected the location of important genes and discovered 18 times more promoters than identified previously. S. Typhimurium...

Identification of a umuDC locus in Salmonella typhimurium LT2

International Nuclear Information System (INIS)

Smith, C.M.; Eisenstadt, E.

1989-01-01

The umuDC operon of Escherichia coli is required for efficient mutagenesis by UV light and many other DNA-damaging agents. The existence of a umuDC analog in Salmonella typhimurium has been questioned. With DNA probes to the E. coli umuD and umuC genes, we detected, by Southern blot hybridization, sequences similar to both of these genes in S. typhimurium LT2. We also confirmed that the presence of cloned E. coli umuD enhances the UV mutability and resistance of S. typhimurium. Our data strongly suggest that S. typhimurium contains a functional umuDC operon

Ascertaining the relationship between Salmonella Typhimurium and Salmonella 4,[5],12:i:- by MLVA and inferring the sources of human salmonellosis due to the two serovars in Italy

DEFF Research Database (Denmark)

Barco, Lisa; Barrucci, Federica; Cortini, Enzo

2015-01-01

The current picture of human salmonellosis shows Salmonella Typhimurium and S. 4,[5],12:i:- as the most common serovars in Italy. The aims of this study were to investigate the genetic relationship between these serovars, as well as to test the possibility of inferring sources of human...... salmonellosis due to S. Typhimurium and S. 4,[5],12:i:- by using multilocus variable-number tandem repeat analysis (MLVA) subtyping data. Single isolates from 268 human sporadic cases and 325 veterinary isolates (from pig, cattle, chicken, and turkey) collected over the period 2009-2011 were typed by MLVA......, and the similarities of MLVA profiles were investigated using different analytical approaches. Results showed that isolates of S. 4,[5],12:i:- were more clonal compared to S. Typhimurium and that clones of both serovars from different non-human sources were very close to those which were responsible for human...

Psoralen photomutagenic specificity in Salmonella typhimurium

International Nuclear Information System (INIS)

Koch, W.H.

1986-01-01

The cytotoxic and mutagenic specificity of two therapeutically employed psoralens was examined in several Ames Salmonella typhimurium strains with near ultraviolet light activation. Photomutagenic activity of 8-methoxypsoralen (8MOP) and 4,5',8-trimethylpsoralen (TMP) was found to be sequence-specific, and additionally was dependent on the level of DNA-repair proficiency. Phototoxicity was essentially identical in hisC3076, hisD3052 and hisG46 strains; uvrB - excision-repair-deficient bacteria were considerably more susceptible to lethal effects than wild-type parental strains. Finally, the data show that psoralens are potent frameshift photomutagens in Salmonella hisC3076 strains and demonstrate the potential utility of these strains in evaluating photomutagenic and phototoxic activity of new furocoumarin derivatives. (Auth.)

Experimental infection of broiler chicks with Salmonella Typhimurium from pigeon (Columba livia

Directory of Open Access Journals (Sweden)

Átilla Holanda de Albuquerque

2016-08-01

Full Text Available Several cases of animal and human salmonellosis caused by the Salmonella serotype Typhimurium have been reported. In animals, subclinical infection favors pathogen dissemination through feces. In this context, the domestic pigeon (Columba livia with an asymptomatic condition may play an important role in the transmission of salmonellosis, through the elimination of contaminated feces in commercial aviaries or in poultry feed facilities, causing economic losses to the poultry industry and presenting a risk to public health. This study aimed to evaluate the mortality, clinical signs and the presence of Salmonella Typhimurium in the feces and organs of chicks previously inoculated with bacteria isolated from a pigeon. One-day-old chicks were distributed in two experimental groups (G1 and G2 of 32 birds each, and a control group of six birds. Two inocula of 0.4 and 0.7 mL with 105 and 106 colony forming units were used in G1 and G2 birds, respectively. At 1, 4, 7 and 14 days post-inoculation (dpi fecal samples were pooled from each cage and individual cloacal swabs were collected. At 14 dpi, all chicks were euthanized and samples were collected from the liver, spleen, lung, cecum and intestine for microbiological analysis. Mortality was only observed among G2 birds (6.25%. Most birds presented clinical signs of diarrhea at 4 dpi and no symptom as observed at 14 dpi. The results from cloacal swabs demonstrated bacterial elimination in 68.8% and 53.1% of G2 and G1 birds, respectively at 1 dpi. Additionally, fecal samples had elevated bacterial shedding in all four periods of observation , with a higher excretion at 4 dpi (62.5% for both groups. Among G2 birds, 74.2% were positive for the pathogen in the intestine; G1 birds presented the lowest rate of lung infection (29%, and both groups had more than 50% positivity for liver and caeca. The results revealed that infected chicks with a Salmonella Typhimurium strains isolated from pigeons may host the

Chlortetracycline and florfenicol induce expression of genes associated with pathogenicity in multidrug-resistant Salmonella enterica serovar Typhimurium

Science.gov (United States)

Background Multidrug-resistant (MDR) Salmonella enterica serovar Typhimurium (S. Typhimurium) is a serious public health threat as infections caused by these strains are more difficult and expensive to treat. Livestock serve as a reservoir for MDR Salmonella, and the antibiotics chlortetracycline an...

Two alanine racemase genes in Salmonella typhimurium that differ in structure and function.

OpenAIRE

Wasserman, S A; Walsh, C T; Botstein, D

1983-01-01

Mutations were isolated in a previously undescribed Salmonella typhimurium gene encoding an alanine racemase essential for utilization of L-alanine as a source of carbon, energy, and nitrogen. This new locus, designated dadB, lies within one kilobase of the D-alanine dehydrogenase locus (dadA), which is also required for alanine catabolism. The dadA and dadB genes are coregulated. Mutants (including insertions) lacking the dadB alanine racemase do not require D-alanine for growth unless a mut...

Salmonella Typhimurium induces SPI-1 and SPI-2 regulated and strain dependent downregulation of MHC II expression on porcine alveolar macrophages

Directory of Open Access Journals (Sweden)

Van Parys Alexander

2012-06-01

Full Text Available Abstract Foodborne salmonellosis is one of the most important bacterial zoonotic diseases worldwide. Salmonella Typhimurium is the serovar most frequently isolated from persistently infected slaughter pigs in Europe. Circumvention of the host’s immune system by Salmonella might contribute to persistent infection of pigs. In the present study, we found that Salmonella Typhimurium strain 112910a specifically downregulated MHC II, but not MHC I, expression on porcine alveolar macrophages in a Salmonella pathogenicity island (SPI-1 and SPI-2 dependent way. Salmonella induced downregulation of MHC II expression and intracellular proliferation of Salmonella in macrophages were significantly impaired after opsonization with Salmonella specific antibodies prior to inoculation. Furthermore, the capacity to downregulate MHC II expression on macrophages differed significantly among Salmonella strains, independently of strain specific differences in invasion capacity, Salmonella induced cytotoxicity and altered macrophage activation status. The fact that strain specific differences in MHC II downregulation did not correlate with the extent of in vitro SPI-1 or SPI-2 gene expression indicates that other factors are involved in MHC II downregulation as well. Since Salmonella strain dependent interference with the pig’s immune response through downregulation of MHC II expression might indicate that certain Salmonella strains are more likely to escape serological detection, our findings are of major interest for Salmonella monitoring programs primarily based on serology.

Genotypic and phenotypic characterization of multidrug resistant Salmonella Typhimurium and Salmonella Kentucky strains recovered from chicken carcasses.

Directory of Open Access Journals (Sweden)

Rizwana Tasmin

Full Text Available Salmonella Typhimurium is the leading cause of human non-typhoidal gastroenteritis in the US. S. Kentucky is one the most commonly recovered serovars from commercially processed poultry carcasses. This study compared the genotypic and phenotypic properties of two Salmonella enterica strains Typhimurium (ST221_31B and Kentucky (SK222_32B recovered from commercially processed chicken carcasses using whole genome sequencing, phenotype characterizations and an intracellular killing assay. Illumina MiSeq platform was used for sequencing of two Salmonella genomes. Phylogenetic analysis employing homologous alignment of a 1,185 non-duplicated protein-coding gene in the Salmonella core genome demonstrated fully resolved bifurcating patterns with varying levels of diversity that separated ST221_31B and SK222_32B genomes into distinct monophyletic serovar clades. Single nucleotide polymorphism (SNP analysis identified 2,432 (ST19 SNPs within 13 Typhimurium genomes including ST221_31B representing Sequence Type ST19 and 650 (ST152 SNPs were detected within 13 Kentucky genomes including SK222_32B representing Sequence Type ST152. In addition to serovar-specific conserved coding sequences, the genomes of ST221_31B and SK222_32B harbor several genomic regions with significant genetic differences. These included phage and phage-like elements, carbon utilization or transport operons, fimbriae operons, putative membrane associated protein-encoding genes, antibiotic resistance genes, siderophore operons, and numerous hypothetical protein-encoding genes. Phenotype microarray results demonstrated that ST221_31B is capable of utilizing certain carbon compounds more efficiently as compared to SK222_3B; namely, 1,2-propanediol, M-inositol, L-threonine, α-D-lactose, D-tagatose, adonitol, formic acid, acetoacetic acid, and L-tartaric acid. ST221_31B survived for 48 h in macrophages, while SK222_32B was mostly eliminated. Further, a 3-fold growth of ST221_31B was

Serovars of Salmonella isolated from Danish turkeys between 1995 and 2000 and their antimicrobial resistance

DEFF Research Database (Denmark)

Pedersen, Karl; Hansen, H.C.; Jørgensen, J.C.

2002-01-01

, florfenicol, or amoxycillin with clavulanic acid, only 24 isolates were resistant to two or more compounds in various combinations of up to six compounds; one Salmonella Havana isolate was resistant to six compounds. Six isolates were serovar Typhimurium, but none of them belonged to phage type DT104....

Identification and Characterization of Outer Membrane Vesicle-Associated Proteins in Salmonella enterica Serovar Typhimurium

Science.gov (United States)

Bai, Jaewoo; Kim, Seul I; Ryu, Sangryeol

2014-01-01

Salmonella enterica serovar Typhimurium is a primary cause of enteric diseases and has acquired a variety of virulence factors during its evolution into a pathogen. Secreted virulence factors interact with commensal flora and host cells and enable Salmonella to survive and thrive in hostile environments. Outer membrane vesicles (OMVs) released from many Gram-negative bacteria function as a mechanism for the secretion of complex mixtures, including virulence factors. We performed a proteomic analysis of OMVs that were isolated under standard laboratory and acidic minimal medium conditions and identified 14 OMV-associated proteins that were observed in the OMV fraction isolated only under the acidic minimal medium conditions, which reproduced the nutrient-deficient intracellular milieu. The inferred roles of these 14 proteins were diverse, including transporter, enzyme, and transcriptional regulator. The absence of these proteins influenced Salmonella survival inside murine macrophages. Eleven of these proteins were predicted to possess secretion signal sequences at their N termini, and three (HupA, GlnH, and PhoN) of the proteins were found to be translocated into the cytoplasm of host cells. The comparative proteomic profiling of OMVs performed in this study revealed different protein compositions in the OMVs isolated under the two different conditions, which indicates that the OMV cargo depends on the growth conditions and provides a deeper insight into how Salmonella utilizes OMVs to adapt to environmental changes. PMID:24935973

Inhibitory Effects of Several Essential Oils towards Salmonella typhimurium, Salmonella paratyphi A and Salmonella paratyphi B

Directory of Open Access Journals (Sweden)

S.F. Mazhar

2014-09-01

Full Text Available Plant essential oils are natural products extracted from plants and because of their antimicrobial properties can be used as natural additives in foods. They are also useful for decontamination of food-borne pathogens and can be a safe additive in foods. The antimicrobial activities of essential oils belonging to Saturiea hortensis, Thymus vulgaris, Mentha polegium, Cuminum cyminum, Lavandula officinalis and Mentha viridis L. (spearmint were investigated at different concentrations (0.1, 0.3, 0.5, 1, 2, 5 and 10%v/v against Salmonella typhimurium, Salmonella paratyphi A and Salmonella paratyphi B by using the agar well diffusion method. Essential oils showed inhibitory effect on Salmonella spp. in the agar well diffusion assay. In addition, the capability of essential oils for decontamination of minced row beef, ground beef, minced raw chicken and minced raw fish inoculated with Salmonella spp. at 0.1 and 0.5%v/v were assessed. Reduction of the Salmonella spp. population was observed following the inoculation of the cultures with 0.1 and 0.5%v/v essential oils.

Genetic Diversity in Salmonella Isolates from Ducks and their Environments in Penang, Malaysia using Enterobacterial Repetitive Intergenic Consensus

Directory of Open Access Journals (Sweden)

Frederick Adzitey 1, Gulam Rusul Rahmat Ali2*, Nurul Huda2 and Rosma Ahmad3

2013-07-01

Full Text Available A total of 107 Salmonella isolates (37 S. typhimurium, 26 S. hadar, 15 S. enteritidis, 15 S. braenderup, and 14 S. albany isolated from ducks and their environments in Penang, Malaysia were typed using enterobacterial repetitive intergenic consensus (ERIC to determine their genetic diversity. Analysis of the Salmonella strains by ERIC produced DNA fingerprints of different sizes for differentiation purposes. The DNA fingerprints or band sizes ranged from 14-8300bp for S. Typhimurium, 146-6593bp for S. hadar, 15-4929bp for S. enteritidis, 14-5142bp for S. braenderup and 7-5712bp for S. albany. Cluster analysis at a coefficient of 0.85 grouped the Salmonella strains into various clusters and singletons. S. typhimurium were grouped into 10 clusters and 6 singletons, S. Hadar were grouped into 3 clusters and 18 singletons, S. enteritidis were grouped into 3 clusters and 7 singletons, S. braenderup were grouped into 4 clusters and 7 singletons, and S. albany were grouped into 3 clusters and 7 singletons with discriminatory index (D ranging from 0.92-0.98. ERIC proved to be a useful typing tool for determining the genetic diversity of the duck Salmonella strains.

Salmonella infection in healthy pet reptiles: Bacteriological isolation and study of some pathogenic characters.

Science.gov (United States)

Bertelloni, Fabrizio; Chemaly, Marianne; Cerri, Domenico; Gall, Françoise Le; Ebani, Valentina Virginia

2016-06-01

The fecal samples from 213 captive reptiles were examined, and 29 (13.61%) Salmonella enterica isolates were detected: 14/62 (22.58%) from chelonians, 14/135 (10.37%) from saurians, and 1/16 (6.25%) from ophidians. The isolates were distributed among 14 different serotypes: Miami, Ebrie, Hermannsweder, Tiergarten, Tornov, Pomona, Poona, Goteborg, Abaetetube, Nyanza, Kumasi, Typhimurium, 50:b:z6, 9,12:z29:1,5, and a non-motile serotype with antigenic formula 1,4,[5],12:-:-. Salmonella typhimurium and 50:b:z6 isolates showed the spv plasmid virulence genes, responsible of the capability to induce extra-intestinal infections. In some cases, pulsed field gel electrophoresis revealed different profiles for the strains of the same serotypes, showing different origins, whereas a common source of infection was supposed when one pulsotype had been observed for isolates of a serovar. Twenty-seven (93.10%) isolates showed resistance to one or more antibiotics. Ceftazidime was active to all the tested isolates, whereas the highest percentages of strains were no susceptible to tigecycline (93.10%), streptomycin (89.66%), and sulfonamide (86.21%).

Inducible pathway is required for mutagenesis in Salmonella typhimurium LT2

International Nuclear Information System (INIS)

Orrego, C.; Eisenstadt, E.

1987-01-01

UV mutability of Salmonella typhimurium LT2 was eliminated in the presence of a multicopy plasmid carrying the Escherichia coli lexA + gene. This result suggests that inducible, SOS-like functions are required for UV mutagenesis in S. typhimurium. S. typhimurium strains carrying either point or deletion mutations in topA had previously been shown to lose their mutability by UV or methyl methanesulfonate. Mitomycin C induction of the Phi(mucB'-lacZ') fusion (a DNA damage-inducible locus carried on plasmid pSE205) in S. typhimurium topA was normal, suggesting that RecA is activated in topA mutants. These observations lead the authors deduce that S. typhimurium has at least one DNA damage-inducible locus in addition to recA that is required for UV mutability

2D proteome analysis initiates new Insights on the Salmonella Typhimurium LuxS protein

Directory of Open Access Journals (Sweden)

Vanderleyden Jos

2009-09-01

Full Text Available Abstract Background Quorum sensing is a term describing a bacterial communication system mediated by the production and recognition of small signaling molecules. The LuxS enzyme, catalyzing the synthesis of AI-2, is conserved in a wide diversity of bacteria. AI-2 has therefore been suggested as an interspecies quorum sensing signal. To investigate the role of endogenous AI-2 in protein expression of the Gram-negative pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium, we performed a 2D-DIGE proteomics experiment comparing total protein extract of wildtype S. Typhimurium with that of a luxS mutant, unable to produce AI-2. Results Differential proteome analysis of wildtype S. Typhimurium versus a luxS mutant revealed relatively few changes beyond the known effect on phase 2 flagellin. However, two highly differentially expressed protein spots with similar molecular weight but differing isoelectric point, were identified as LuxS whereas the S. Typhimurium genome contains only one luxS gene. This observation was further explored and we show that the S. Typhimurium LuxS protein can undergo posttranslational modification at a catalytic cysteine residue. Additionally, by constructing LuxS-βla and LuxS-PhoA fusion proteins, we demonstrate that S. Typhimurium LuxS can substitute the cognate signal peptide sequences of β-lactamase and alkaline phosphatase for translocation across the cytoplasmic membrane in S. Typhimurium. This was further confirmed by fractionation of S. Typhimurium protein extracts, followed by Western blot analysis. Conclusion 2D-DIGE analysis of a luxS mutant vs. wildtype Salmonella Typhimurium did not reveal new insights into the role of AI-2/LuxS in Salmonella as only a small amount of proteins were differentially expressed. However, subsequent in depth analysis of the LuxS protein itself revealed two interesting features: posttranslational modification and potential translocation across the cytoplasmic membrane. As

Structural characterization of the Salmonella typhimurium LT2 umu operon

International Nuclear Information System (INIS)

Thomas, S.M.; Crowne, H.M.; Pidsley, S.C.; Sedgwick, S.G.

1990-01-01

The umuDC operon of Escherichia coli encodes functions required for mutagenesis induced by radiation and a wide variety of chemicals. The closely related organism Salmonella typhimurium is markedly less mutable than E. coli, but a umu homolog has recently been identified and cloned from the LT2 subline. In this study the nucleotide sequence and structure of the S. typhimurium LT2 umu operon have been determined and its gene products have been identified so that the molecular basis of umu activity might be understood more fully. S. typhimurium LT2 umu consists of a smaller 417-base-pair (bp) umuD gene ending 2 bp upstream of a larger 1,266-bp umuC gene. The only apparent structural difference between the two operons is the lack of gene overlap. An SOS box identical to that found in E. coli is present in the promoter region upstream of umuD. The calculated molecular masses of the umuD and umuC gene products were 15.3 and 47.8 kilodaltons, respectively, which agree with figures determined by transpositional disruption and maxicell analysis. The S. typhimurium and E. coli umuD sequences were 68% homologous and encoded products with 71% amino acid identity; the umuC sequences were 71% homologous and encoded products with 83% amino acid identity. Furthermore, the potential UmuD cleavage site and associated catalytic sites could be identified. Thus the very different mutagenic responses of S. typhimurium LT2 and E. coli cannot be accounted for by gross differences in operon structure or gene products. Rather, the ability of the cloned S. typhimurium umuD gene to give stronger complementation of E. coli umuD77 mutants in the absence of a functional umuC gene suggests that Salmonella UmuC protein normally constrains UmuD protein activity

Cloning of Salmonella typhimurium DNA encoding mutagenic DNA repair

International Nuclear Information System (INIS)

Thomas, S.M.; Sedgwick, S.G.

1989-01-01

Mutagenic DNA repair in Escherichia coli is encoded by the umuDC operon. Salmonella typhimurium DNA which has homology with E. coli umuC and is able to complement E. coli umuC122::Tn5 and umuC36 mutations has been cloned. Complementation of umuD44 mutants and hybridization with E. coli umuD also occurred, but these activities were much weaker than with umuC. Restriction enzyme mapping indicated that the composition of the cloned fragment is different from the E. coli umuDC operon. Therefore, a umu-like function of S. typhimurium has been found; the phenotype of this function is weaker than that of its E. coli counterpart, which is consistent with the weak mutagenic response of S. typhimurium to UV compared with the response in E. coli

Characterisation of recently emerged multiple antibiotic-resistant Salmonella enterica serovar typhimurium DT104 and other multiresistant phage types from Danish pig herds

DEFF Research Database (Denmark)

Baggesen, Dorte Lau; Aarestrup, Frank Møller

1998-01-01

electrophoresis (PFGE) using the restriction enzyme Xba I, Overall, 66 per cent of the 670 isolates were sensitive to all the antimicrobial agents tested. Eleven isolates of S typhimurium were resistant to ampicillin, streptomycin and tetracycline and also resistant to other antibiotics in different resistance...... patterns. Seven different multiresistant clones were identified, The most common clones were four isolates of DT104 and three isolates of DT193, TWO Of the three S typhimurium DT104 from 1994 and 1995 were sensitive to all the antimicrobials tested whereas the remaining isolate from 1994 was resistant......A total of 670 isolates of Salmonella enterica were isolated from Danish pig herds, phage typed and tested for susceptibility to amoxycillin + clavulanate, ampicillin, colistin, enrofloxacin, gentamicin, neomycin, spectinomycin, streptomycin, tetracyclines, and trimethoprim + sulphadiazine. S...

IDENTIFICATION OF SALMONELLA SEROVARS ISOLATED DURING 2009-2016 IN TERNOPIL REGION, UKRAINE

Directory of Open Access Journals (Sweden)

Pokryshko O.V.

2017-06-01

Full Text Available Introduction. Salmonellosis is registered in all regions of the world. Relevance of salmonellosis is due its global distribution, increasing incidence, even in developed countries, frequent outbreaks. The most reports in different countries demonstrated that one of the common Salmonella serotypes isolated from food and environmental samples had been serovars Salmonella Enterica, Typhimurium. In Ukraine 7.3% of all acute diarrheal infections have been cases of salmonellosis. Although large Salmonella outbreaks usually attract media attention, 60–80% of all salmonellosis cases are not recognized as part of a known outbreak and are classified as sporadic cases, or are not diagnosed as such at all. Material & methods. The samples from cultured stool, bile samples, food and environment were inoculated in the Tryptic Soya Broth (TSB for the enrichment and detection of the bacteria. After 24 hours incubation, microorganisms were cultured on the MacConkey agar plates. Then biochemical and serological tests were performed to identify the serovars of the isolated Salmonella in Ternopil regional laboratory center, Ukraine.Results & discussion. Over the past 8 years the incidence of salmonellosis has varied between 8.41 3.3 cases per 100 thousand of population (35 - 90 cases. During this period, the lowest rate recorded in 2015 (3.3 cases per 100 thousand of population, the highest – in 2014. Analysis of morbidity has been shown that elevated levels of infection were due to outbreaks registrated in 2011 (the number of infected people was 23, in 2013 (53 infected people, in 2014 (67 infected people and in 2016 (16 infected people. In Ternopil region the dominant serovar of Salmonella spp. isolated from patients are S. enteritidis (56.8 - 93.5% of all cases of diseases and S. typhimurium (7.8 - 43.8% in last 8 years. Among the carriers circulate S.enteritidis, S. typhimurium – mainly (64,8% and 35.2% respectively. Not typical for Ternopil region

Salmonella typhimurium in the hip: look before a SLE patient leaps.

Science.gov (United States)

Kumar, V Anil; Krishna, Sushma; Thilak, Jai; Dinesh, Kavitha R; Karim, Shamsul

2011-07-01

Salmonellosis is a common infection in systemic lupus erythematosus patients. The present study reports on two such cases of Salmonella typhimurium septic arthritis among the 102 patients seen during a four-year period.

Thermal inactivation of Salmonella Typhimurium in chicken shawirma (gyro).

Science.gov (United States)

Osaili, Tareq M; Al-Nabulsi, Anas A; Shaker, Reyad R; Olaimat, Amin N; Jaradat, Ziad W; Holley, Richard A

2013-08-16

This study explored the thermal characteristics (D- and z-values) of Salmonella Typhimurium in raw chicken shawirma. Marinated and non-marinated chicken breasts with skin were inoculated with S. Typhimurium 112 or S. Typhimurium 144. Inoculated samples were ground, packed in sterile bags and submerged in a water bath at 54, 56, 58 and 60°C for 2.5 to 72min. The mean D-values of S. Typhimurium strains in inoculated, non-marinated, ground raw chicken breast, as well as those of S. Typhimurium 15h after exposure to the marinade (inoculated before marinating, IBM) or after brief exposure (30min) to the marinade (inoculated after marinating, IAM) ranged from 9.15 to 12.44, 2.89 to 3.92, 1.06 to 1.30 and 0.32 to 0.52min at 54, 56, 58 and 60°C, respectively. Generally, no significant differences (P>0.05) were found among the D-values of S. Typhimurium in all chicken samples. However, the D-values of S. Typhimurium in raw ground chicken shawirma IBM were the lowest. The z-values of S. Typhimurium in all products ranged from 3.78 to 4.58°C. It was concluded that thorough cooking of the outside of the shawirma meat cylinder or cone before removal of slices at foodservice counters can enhance the safety of the product. Copyright © 2013 Elsevier B.V. All rights reserved.

Prevalence, serovars, phage types, and antibiotic susceptibilities of Salmonella strains isolated from animals in the United Arab Emirates from 1996 to 2009.

Science.gov (United States)

Münch, Sebastian; Braun, Peggy; Wernery, Ulrich; Kinne, Jörg; Pees, Michael; Flieger, Antje; Tietze, Erhard; Rabsch, Wolfgang

2012-10-01

The aim of this study was to give some insights into the prevalence, serovars, phage types, and antibiotic resistances of Salmonella from animal origin in the United Arab Emirates. Data on diagnostic samples from animals (n = 20,871) examined for Salmonella between 1996 and 2009 were extracted from the databases of the Central Veterinary Research Laboratory in Dubai and from typed strains (n = 1052) from the Robert Koch Institute, Wernigerode Branch in Germany and analyzed for general and animal-specific trends. Salmonella was isolated from 1,928 (9 %) of the 20,871 samples examined. Among the 1,052 typed strains, most were from camels (n = 232), falcons (n = 166), bustards (n = 101), antelopes (n = 66), and horses (n = 63). The predominant serovars were Salmonella Typhimurium (25 %), Salmonella Kentucky (8 %), followed by Salmonella Frintrop (7 %), and Salmonella Hindmarsh (5 %). When analyzed by animal species, the most frequent serovars in camels were Salmonella Frintrop (28 %) and Salmonella Hindmarsh (21 %), in falcons Salmonella Typhimurium (32 %), in bustards Salmonella Kentucky (19 %), in antelopes Salmonella Typhimurium (9 %), and in horses Salmonella Typhimurium (17 %) and S. Kentucky (16 %). Resistance of all typed Salmonella strains (n = 1052) was most often seen to tetracycline (23 %), streptomycin (22 %), nalidixic acid (18 %), and ampicillin (15 %). These data show trends in the epidemiology of Salmonella in different animal species which can be used as a base for future prevention, control, and therapy strategies.

Large outbreaks of Salmonella Typhimurium infection in Denmark in 2008

DEFF Research Database (Denmark)

Ethelberg, S.; Wingstrand, Anne; Jensen, T.

2008-01-01

An outbreak of Salmonella Typhimurium phage type U292 has been ongoing in Denmark since 1 April, with 1,054 cases registered until 23 October 2008. Extensive investigations including hypothesis-generating interviews, matched case-control studies, cohort studies in embedded outbreaks, shopping list...

Identification of Salmonella serovars isolated from live molluscan shellfish and their significance in the marine environment.

Science.gov (United States)

Martinez-Urtaza, Jaime; Saco, Montserrat; Hernandez-Cordova, Gustavo; Lozano, Antonio; Garcia-Martin, Oscar; Espinosa, Joaquin

2003-02-01

A study on the presence of Salmonella spp. in live molluscs was performed, which included a description of the different serovars isolated and their relationship to the marine environment. A total of 2,980 samples of shellfish from Galicia (N.W. Spain) were tested for the presence of Salmonella spp. between September 1998 and August 2001. The overall incidence of Salmonella was 1.8% and showed a slight rise during the 3 years of the study. Mussels and oysters presented a higher incidence than clams and cockles, possibly because of their distinct growing habitat. A seasonal pattern was noted for the isolation of Salmonella spp.: 54% of the isolations were detected from September to November. That nearly 67% of the total Salmonella was isolated from shellfish with fecal coliform levels fecal coliforms do not necessarily indicate the absence of Salmonella. A total of nine serovars were found in the 54 Salmonella isolated. Salmonella Senftenberg was the most frequent (50%), followed by Salmonella Typhimurium (18%) and Salmonella Agona (17%). Salmonella Senftenberg was detected frequently during the year, whereas the remaining serovars were detected only on occasional contamination events.

Multidrug resistance among different serotypes of clinical Salmonella isolates in Taiwan

DEFF Research Database (Denmark)

Lauderdale, T. L.; Aarestrup, Frank Møller; Chen, P. C.

2006-01-01

(41%) and was highly prevalent in Salmonella enterica serotype Typhimurium (72.7%, 176/242) the most common serotype. Additional resistance to trimethoprim was present in 155 (19.4% overall) of the ACSSuT R-type isolates from several serotypes. Reduced susceptibility to fluoroquinolone (FQ...... multiresistant to other antimicrobials. Studies are needed to determine the sources of different multidrug-resistant serotypes. Continued national surveillance is underway to monitor changes in resistance trends and to detect further emergence of resistant Salmonella serotypes in Taiwan. (c) 2006 Elsevier Inc...

Buffer capacity of food components influences the acid tolerance response in Salmonella Typhimurium during simulated gastric passage

DEFF Research Database (Denmark)

Aabo, Søren; Buschhardt, Tasja; Hansen, Tina Beck

2014-01-01

Food composition, buffer capacity, and fat and protein content have been shown to effect the gastric acid survival of pathogens (Waterman & Small 1998). In this study, simple food-model substances with different buffer capacities were investigated for their ability to support survival of stationary...... Heart Infusion Broth having a higher buffer capacity. We suggest this to be associated with a varying ability of Salmonella Typhimurium to mount a stationary phase acid tolerance response (ATR) depending on the buffer capacity of the food vehicle....... phase Salmonella Typhimurium during simulated gastric acid passage. We used a computer-controlled fermentor to employ pH changes in synthetic gastric fluid, mimicking the dynamic pH during gastric passage. In order to minimise variation, Salmonella enterica serovar Typhimurium was contained in dialysis...

Polyamines Are Required for Virulence in Salmonella enterica Serovar Typhimurium

DEFF Research Database (Denmark)

Jelsbak, Lotte; Thomsen, Line Elnif; Wallrodt, Inke

2012-01-01

for studying typhoid fever. Central to its virulence are two major virulence loci Salmonella Pathogenicity Island 1 and 2 (SPI1 and SPI2). SPI1 promotes invasion of epithelial cells, whereas SPI2 enables S. Typhimurium to survive and proliferate within specialized compartments inside host cells. In this study......, we show that an S. Typhimurium polyamine mutant is defective for invasion, intracellular survival, killing of the nematode Caenorhabditis elegans and systemic infection of the mouse model of typhoid fever. Virulence of the mutant could be restored by genetic complementation, and invasion...

Host Specificity of Salmonella typhimurium Deoxyribonucleic Acid Restriction and Modification

Science.gov (United States)

Slocum, Harvey; Boyer, Herbert W.

1973-01-01

The restriction and modification genes of Salmonella typhimurium which lie near the thr locus were transferred to a restrictionless mutant of Escherichia coli. These genes were found to be allelic to the E. coli K, B, and A restriction and modification genes. E. coli recombinants with the restriction and modification host specificity of S. typhimurium restricted phage λ that had been modified by each of the seven known host specificities of E. coli at efficiency of plating levels of about 10−2. Phage λ modified with the S. typhimurium host specificity was restricted by six of the seven E. coli host specificities but not by the RII (fi− R-factor controlled) host specificity. It is proposed that the restriction and modification enzymes of this S. typhimurium host specificity have two substrates, one of which is a substrate for the RII host specificity enzymes. PMID:4570605

Salmonella Species' Persistence and Their High Level of Antimicrobial Resistance in Flooded Man-Made Rivers in China.

Science.gov (United States)

Song, Qifa; Zhang, Danyang; Gao, Hong; Wu, Junhua

2018-05-11

Man-made rivers, owing to proximity to human habitats, facilitate transmission of salmonellosis to humans. To determine the contamination situation by Salmonella in flooded man-made rivers and thereafter the exposure risk to public health, we investigated the prevalence of Salmonella species and their antimicrobial resistance in such rivers, as well as the relationship between the incidence of local infectious diarrhea cases and the number of Salmonella isolates from patients. After a heavy flood, 95 isolates of 13 Salmonella serotypes were isolated from 80 river water samples. The two most prevalent serotypes were Typhimurium and Derby. Eight Salmonella serotypes were newly detected after the flood. Overall, 50 isolates were resistant to ampicillin and/or cefotaxime and carried at least bla TEM . Twelve isolates of serotypes Typhimurium, Derby, Rissen, and Indiana were extended-spectrum β-lactamase (ESBL) producing and carried at least one of bla OXA and bla CTX-M-like genes. Twelve isolates of serotypes Typhimurium, Derby, Agona, Rissen, and Indiana were resistant to ciprofloxacin and had gyrA mutations. Isolates of Typhimurium, Derby, and Indiana were concurrently ciprofloxacin resistant and ESBL producing. Pulsed-field gel electrophoresis illustrates the circulation of two dominant clones of Salmonella Typhimurium isolates among patients, river, and food. High prevalence of various highly pathogenic and antimicrobial-resistant Salmonella serotypes shows that man-made rivers are prone to heavy contamination with Salmonella, and as a result put public health at greater risk.

Pyridine nucleotide cycle of Salmonella typhimurium: isolation and characterization of pncA, pncB, and pncC mutants and utilization of exogenous nicotinamide adenine dinucleotide.

Science.gov (United States)

Foster, J W; Kinney, D M; Moat, A G

1979-03-01

Mutants of Salmonella typhimurium LT-2 deficient in nicotinamidase activity (pncA) or nicotinic acid phosphoribosyltransferase activity (pncB) were isolated as resistant to analogs of nicotinic acid and nicotinamide. Information obtained from interrupted mating experiments placed the pncA gene at 27 units and the pncB gene at 25 units on the S. typhimurium LT-2 linkage map. A major difference in the location of the pncA gene was found between the S. typhimurium and Escherichia coli linkage maps. The pncA gene is located in a region in which there is a major inversion of the gene order in S. typhimurium as compared to that in E. coli. Growth experiments using double mutants blocked in the de novo pathway to nicotinamide adenine dinucleotide (NAD) (nad) and in the pyridine nucleotide cycle (pnc) at either the pncA or pncB locus, or both, have provided evidence for the existence of an alternate recycling pathway in this organism. Mutants lacking this alternate cycle, pncC, have been isolated and mapped via cotransduction at 0 units. Utilization of exogenous NAD was examined through the use of [14C]carbonyl-labeled NAD and [14C]adenine-labeled NAD. The results of these experiments suggest that NAD is degraded to nicotinamide mononucleotide at the cell surface. A portion of this extracellular nicotinamide mononucleotide is then transported across the cell membrane by nicotinamide mononucleotide glycohydrolase and degraded to nicotinamide in the process. The remaining nicotinamide mononucleotide accumulates extracellularly and will support the growth of nadA pncB mutants which cannot utilize the nicotinamide resulting from the major pathway of NAD degradation. A model is presented for the utilization of exogenous NAD by S. typhimurium LT-2.

Comparative genome analysis and characterization of the Salmonella Typhimurium strain CCRJ_26 isolated from swine carcasses using whole-genome sequencing approach.

Science.gov (United States)

Panzenhagen, P H N; Cabral, C C; Suffys, P N; Franco, R M; Rodrigues, D P; Conte-Junior, C A

2018-04-01

Salmonella pathogenicity relies on virulence factors many of which are clustered within the Salmonella pathogenicity islands. Salmonella also harbours mobile genetic elements such as virulence plasmids, prophage-like elements and antimicrobial resistance genes which can contribute to increase its pathogenicity. Here, we have genetically characterized a selected S. Typhimurium strain (CCRJ_26) from our previous study with Multiple Drugs Resistant profile and high-frequency PFGE clonal profile which apparently persists in the pork production centre of Rio de Janeiro State, Brazil. By whole-genome sequencing, we described the strain's genome virulent content and characterized the repertoire of bacterial plasmids, antibiotic resistance genes and prophage-like elements. Here, we have shown evidence that strain CCRJ_26 genome possible represent a virulence-associated phenotype which may be potentially virulent in human infection. Whole-genome sequencing technologies are still costly and remain underexplored for applied microbiology in Brazil. Hence, this genomic description of S. Typhimurium strain CCRJ_26 will provide help in future molecular epidemiological studies. The analysis described here reveals a quick and useful pipeline for bacterial virulence characterization using whole-genome sequencing approach. © 2018 The Society for Applied Microbiology.

Salmonella spp. in raw broiler parts: occurrence, antimicrobial resistance profile and phage typing of the Salmonella Enteritidis isolates Salmonella spp. em cortes de frango: ocorrência, resistência antimicrobiana e fagotipificação dos isolados de Salmonella Enteritidis

Directory of Open Access Journals (Sweden)

Aldemir Reginato Ribeiro

2007-06-01

Full Text Available The present study was carried out to evaluate the occurrence of Salmonellae in raw broiler parts and to determine the antimicrobial resistance profile of the isolated strains. Twenty-four (39.3% broiler parts samples were positive for Salmonella and twenty-five Salmonella strains were isolated, since two different serovars were detected in one single positive sample. Salmonella Enteritidis was the most prevalent serovar. Among Salmonella Enteritidis isolates, 95.2% belonged to Phage Type 4 (PT4 (20/21 and 4.8% to PT7 (1/21. Twenty-two (88% strains of Salmonella were resistant to at least one antimicrobial agent, generating eight different resistance patterns. The S. Typhimurium (n: 1 and S. Hadar (n: 3 isolates presented multiple resistance. Three S. Enteritidis isolates were susceptible to all antimicrobials tested, two were resistant only to tetracycline. The high prevalence of Salmonella in the broiler parts strenghtens the importance of the use of good manufacturing practices (GMP, and HACCP. The results also emphasize the need for the responsible use of antimicrobials in animal production.Este trabalho foi conduzido para avaliar a ocorrência de Salmonella em cortes de frango e para determinar o perfil de resistência antimicrobiana das cepas isoladas. Vinte e quatro (39,3% cortes de frango foram positivas para Salmonella, tendo sido isoladas vinte e cinco cepas de Salmonella, uma vez que em uma amostra isolaram-se dois sorovares. Salmonella Enteritidis foi o sorovar prevalente. Entre as Salmonella Enteritidis isoladas, 95,2% pertencem ao Fagotipo 4 (PT4 (20/21 e 4,8% ao PT7 (1/21. Vinte e duas (88% cepas de Salmonella foram resistentes a pelo menos um agente antimicrobiano e oito diferentes padrões de resistência foram observados. S. Typhimurium (n:1 e S. Hadar (n: 3, apresentaram múltipla resistência. Três cepas de S. Enteritidis foram sensíveis a todos os antimicrobianos e duas resistentes somente a tetraciclina. A elevada ocorr

A multiplex real-time PCR assay targeting virulence and resistance genes in Salmonella enterica serotype Typhimurium

Directory of Open Access Journals (Sweden)

Brisabois Anne

2011-06-01

Full Text Available Abstract Background Typhimurium is the main serotype of Salmonella enterica subsp. enterica implicated in food-borne diseases worldwide. This study aimed to detect the prevalence of ten markers combined in a macro-array based on multiplex real-time PCR. We targeted characteristic determinants located on pathogenicity islands (SPI-2 to -5, virulence plasmid pSLT and Salmonella genomic island 1 (SGI1 as well as a specific 16S-23S rRNA intergenic spacer sequence of definitive type 104 (DT104. To investigate antimicrobial resistance, the study also targeted the presence of genes involved in sulfonamide (sul1 and beta-lactam (blaTEM resistance. Finally, the intI1 determinant encoding integrase from class 1 integron was also investigated. Results A total of 538 unrelated S. Typhimurium strains isolated between 1999 and 2009 from various sources, including food animals, food products, human and environmental samples were studied. Based on the combined presence or absence of these markers, we distinguished 34 different genotypes, including three major genotypes encountered in 75% of the studied strains, Although SPI determinants were almost always detected, SGI1, intI1, sul1 and blaTEM determinants were found 47%, 52%, 54% and 12% of the time respectively, varying according to isolation source. Low-marker patterns were most often detected in poultry sources whereas full-marker patterns were observed in pig, cattle and human sources. Conclusion The GeneDisc® assay developed in this study madeit easier to explore variability within serotype Typhimurium by analyzing ten relevant gene determinants in a large collection of strains. This real-time multiplex method constitutes a valuable tool for strains characterization on epidemiological purposes.

Inorganic Polyphosphate Is Essential for Salmonella Typhimurium Virulence and Survival in Dictyostelium discoideum

Directory of Open Access Journals (Sweden)

Macarena A. Varas

2018-01-01

Full Text Available Inorganic polyphosphate (polyP deficiency in enteric bacterial pathogens reduces their ability to invade and establish systemic infections in different hosts. For instance, inactivation of the polyP kinase gene (ppk encoding the enzyme responsible for polyP biosynthesis reduces invasiveness and intracellular survival of Salmonella enterica serovar Typhimurium (S. Typhimurium in epithelial cells and macrophages in vitro. In addition, the virulence in vivo of a S. Typhimurium Δppk mutant is significantly reduced in a murine infection model. In spite of these observations, the role played by polyP during the Salmonella-host interaction is not well understood. The social amoeba Dictyostelium discoideum has proven to be a useful model for studying relevant aspects of the host-pathogen interaction. In fact, many intracellular pathogens can survive within D. discoideum cells using molecular mechanisms also required to survive within macrophages. Recently, we established that S. Typhimurium is able to survive intracellularly in D. discoideum and identified relevant genes linked to virulence that are crucial for this process. The aim of this study was to determine the effect of a polyP deficiency in S. Typhimurium during its interaction with D. discoideum. To do this, we evaluated the intracellular survival of wild-type and Δppk strains of S. Typhimurium in D. discoideum and the ability of these strains to delay the social development of the amoeba. In contrast to the wild-type strain, the Δppk mutant was unable to survive intracellularly in D. discoideum and enabled the social development of the amoeba. Both phenotypes were complemented using a plasmid carrying a copy of the ppk gene. Next, we simultaneously evaluated the proteomic response of both S. Typhimurium and D. discoideum during host-pathogen interaction via global proteomic profiling. The analysis of our results allowed the identification of novel molecular signatures that give insight into

Curcumin increases the pathogenicity of Salmonella enterica serovar Typhimurium in murine model.

Directory of Open Access Journals (Sweden)

Sandhya A Marathe

Full Text Available Curcumin has gained immense importance for its vast therapeutic and prophylactic applications. Contrary to this, our study reveals that it regulates the defense pathways of Salmonella enterica serovar Typhimurium (S. Typhimurium to enhance its pathogenicity. In a murine model of typhoid fever, we observed higher bacterial load in Peyer's patches, mesenteric lymph node, spleen and liver, when infected with curcumin-treated Salmonella. Curcumin increased the resistance of S. Typhimurium against antimicrobial agents like antimicrobial peptides, reactive oxygen and nitrogen species. This increased tolerance might be attributed to the up-regulation of genes involved in resistance against antimicrobial peptides--pmrD and pmrHFIJKLM and genes with antioxidant function--mntH, sodA and sitA. We implicate that iron chelation property of curcumin have a role in regulating mntH and sitA. Interestingly, we see that the curcumin-mediated modulation of pmr genes is through the PhoPQ regulatory system. Curcumin downregulates SPI1 genes, required for entry into epithelial cells and upregulates SPI2 genes required to intracellular survival. Since it is known that the SPI1 and SPI2 system can be regulated by the PhoPQ system, this common regulator could explain curcumin's mode of action. This data urges us to rethink the indiscriminate use of curcumin especially during Salmonella outbreaks.

A novel imageable therapeutic probe for cancer; cytolysin a expressing attenuated salmonella typhimurium

Energy Technology Data Exchange (ETDEWEB)

Nguyen, Vu Hong; Tae, Seong Ho; Piao, Hong Hua; Hong, Yeoung Jin; Choy, Hyon E.; Bom, Hee Seung; Min, Jung Joon [Chonnam National University Medical School, Gwangju (Korea, Republic of)

2007-07-01

Oncolytic strategy using bacteria has a long history. With the discovery of fluorescent and luminescent reporter genes, bacteria can be easily monitored continuously in treatment process. Salmonella typhimurium ppGpp mutant, one of the prominent attenuated bacteria, has just reported recently, Therefore, in this study, we established strain Cytolysin A (Cly A) expressing light-emitting S. typhimurium ppGpp mutant. S. typhimurium ppGpp mutant was transducted by lux gene for in vivo imaging (S. typhimurium ppGpp/lux) and then, plasmid containing ClyA gene, which is encoded for a pore-forming protein toxin, was transformed to create the strain expressing haemolytic activity (S. typhimurium ppGpp/lux/ClyA). The toxicity of ClyA was evaluated in vitro by inoculating the bacteria with various cultured cancer cell lines. On the other hand, to test the therapeutic effect, the bacteria were injected intermittently, intraperitoneal y or intravenously into CT26-bearing Balb/c mice. The sizes of tumors were measured and in vivo imaging was taken everyday by IVIS machine (Xenogen). The in vitro result showed the number of death cells were significantly higher in the samples containing S. typhimurium ppGpp/lux/ClyA compared with the samples containing S. typhimurium ppGpp/lux. After two days injection, the growth of tumors were repressed in mice injected with either S. typhimurium ppGpp/lux/ClyA or S. typhimurium ppGpp/lux, while tumors in control group still grew fast. In day 3, the tumors inoculated with S. typhimurium ppGpp/lux/ClyA became necrosis and regressed in the following days but not in other groups. In addition, in vivo imaging data showed that the Salmonella strains selectively located in the tumor. By in vivo imaging technique, the light-emitting bacteria can be easily monitored and quantified non-invasively and repeatedly. And ClyA expressing light-emitting S. typhimurium ppGpp mutant can become an effective and safely candidate for cancer treatment.

Subtyping of Salmonella enterica isolated from humans and food animals using Pulsed-Field Gel Electrophoresis

Directory of Open Access Journals (Sweden)

Golab, N.

2016-07-01

Full Text Available Salmonella infections are the second leading cause of zoonotic bacterial foodborne illness. Main source of infection in human is contaminated food products. The aim of this study was sub typing isolates of Salmonella enterica obtained during our previous study by Pulsed Field Gel Electrophoresis (PFGE technique. All 46 Salmonella isolates were serotyped and then subjected to PFGE. Total isolates were analyzed by means of the molecular technique XbaI PFGE. In this study, PFGE and serotyping were used to subtype 46 Salmonella isolates belonging to 27different serovars and derived from human and different food origins. Among these isolates, S. Typhimurium was found to be the most predominant serovar. 40 PFGE patterns out of 46 isolates were obtained. The Discrimination Index obtained by serotyping (DI = 0.93 was lower than PFGE (DI = 0.99. Subtyping of Salmonella enterica is very important and shows that animal origin can be one of a reservoir that potentially could be transferred to human through the food chain. In addition, results of this study also revealed that this procedure is a golden standard for genotyping of such salmonella serotypes.

Salmonella Typhimurium and multidirectional communication in the gut

Directory of Open Access Journals (Sweden)

Elena V. Gart

2016-11-01

Full Text Available The mammalian digestive tract is home to trillions of microbes, including bacteria, archaea, protozoa, fungi and viruses. In monogastric mammals the stomach and small intestine harbor diverse bacterial populations but are typically less populated than the colon. The gut bacterial community (microbiota hereafter varies widely among different host species and individuals within a species. It is influenced by season of the year, age of the host, stress and disease. Ideally, the host and microbiota benefit each other. The host provides nutrients to the microbiota and the microbiota assists the host with digestion and nutrient metabolism. The resident microbiota competes with pathogens for space and nutrients and, through this competition, protects the host in a phenomenon called colonization resistance. The microbiota participates in development of the host immune system, particularly regulation of autoimmunity and mucosal immune response. The microbiota also shapes gut-brain communication and host responses to stress; and, indeed, the microbiota is a newly recognized endocrine organ within mammalian hosts.Salmonella enterica serovar Typhimurium (S. Typhimurium hereafter is a food-borne pathogen which adapts to and alters the gastrointestinal (GI environment. In the GI tract, S. Typhimurium competes with the microbiota for nutrients and overcomes colonization resistance to establish infection. To do this, S. Typhimurium uses multiple defense mechanisms to resist environmental stressors, like the acidic pH of the stomach, and virulence mechanisms which allow it to invade the intestinal epithelium and disseminate throughout the host. To coordinate gene expression and disrupt signaling within the microbiota and between host and microbiota, S. Typhimurium employs its own chemical signaling and may regulate host hormone metabolism.This review will discuss the multidirectional interaction between S. Typhimurium, host and microbiota as well as mechanisms

Prevalence, Virulence Genes and Antimicrobial Resistance Profiles of Salmonella Serovars from Retail Beef in Selangor, Malaysia

Directory of Open Access Journals (Sweden)

Tze Y. Thung

2018-01-01

Full Text Available The aim of the present study was to investigate the prevalence of Salmonella spp., Salmonella Enteritidis and Salmonella Typhimurium in retail beef from different retail markets of Selangor area, as well as, to assess their pathogenic potential and antimicrobial resistance. A total of 240 retail beef meat samples (chuck = 60; rib = 60; round = 60; sirloin = 60 were randomly collected. The multiplex polymerase chain reaction (mPCR in combination with the most probable number (MPN method was employed to detect Salmonella spp., S. Enteritidis and S. Typhimurium in the meat samples. The prevalence of Salmonella spp., S. Enteritidis and S. Typhimurium in 240 beef meat samples were 7.50, 1.25, and 0.83%, respectively. The microbial loads of total Salmonella was found in the range of <3 to 15 MPN/g. Eight different serovars of Salmonella were identified among the 23 isolates, and S. Agona was the predominant serovar (26.09%. Interestingly, all the Salmonella isolates were resistant to penicillin, erythromycin and vancomycin, but the sensitivity was observed for tetracycline, gentamicin and amoxicillin/clavulanic acid. All 23 isolates were resistant to at least three antibiotics. Two S. Typhimurium isolates (8.70% exhibited the highest multiple antibiotic resistance (MAR index value of 0.56 which shown resistance to nine antibiotics. PCR analysis of virulence genes showed that all Salmonella isolates (100% were positive for the invA gene. Meanwhile, pefA was only identified in S. Enteritidis and S. Typhimurium. The findings in this study indicate that retail beef products tested were widely contaminated with multi-drug resistant (MDR Salmonella and various virulence genes are present among the isolated Salmonella serovars.

Sources of salmonellae in an uninfected commercially-processed broiler flock.

Science.gov (United States)

Rigby, C E; Pettit, J R; Baker, M F; Bentley, A H; Salomons, M O; Lior, H

1980-07-01

Cultural monitoring was used to study the incidence and sources of salmonellae in a 4160 bird broiler flock during the growing period, transport and processing in a commercial plant. No salmonellae were isolated from any of 132 litter samples of 189 chickens cultured during the seven-week growing period, even though nest litter samples from four of the eight parent flocks yielded salmonellae and Salmonella worthington was isolated from the meat meal component of the grower ration. On arrival at the plant, 2/23 birds sampled carried S. infantis on their feathers, although intestinal cultures failed to yield salmonellae. Three of 18 processed carcasses samples yielded salmonellae (S. infantis, S. heidelberg, S. typhimurium var copenhagen). The most likely source of these salmonellae was the plastic transport crates, since 15/107 sampled before the birds were loaded yielded salmonellae (S. infantis, S. typhimurium). The crate washer at the plant did not reduce the incidence of Salmonella-contaminated crates, since 16/116 sampled after washing yielded salmonellae (S. infantis, S. typhimurium, S. heidelberg, S. schwarzengrund, S. albany).

Position on mouse chromosome 1 of a gene that controls resistance to Salmonella typhimurium.

Science.gov (United States)

Taylor, B A; O'Brien, A D

1982-06-01

Ity is a gene which regulates the magnitude of Salmonella typhimurium growth in murine tissues and, hence, the innate salmonella resistance of mice. The results of a five-point backcross clearly showed that the correct gene order on chromosome 1 is fz-Idh-1-Ity-ln-Pep-3.

Liver function and bacteriology of organs in broiler inoculated with nalidixic acid-resistant Salmonella Typhimurium and treated with organic acids

Directory of Open Access Journals (Sweden)

Tatiane M. Rocha

2013-07-01

Full Text Available AbAns etxrpaecritment was carried out with 630 one-day-old chicks to evaluate the effects of organic acids when birds were experimentally inoculated with Salmonella Typhimurium. Liver damage and the persistence of the bacterium in the organs were evaluated as well. Broilers were distributed in a completely randomised experimental design in a 3×2 factorial arrangement of six treatments with seven replicates of 15 birds each. Birds were inoculated with saline solution or the bacterium via gavage at 1 day of age, or were offered a feed containing or not the organic acid blend for the period of 7 to 14 days of age. A dose of 5.0x102 colony-forming units (CFU/0.5 mL of Salmonella Typhimurium was used for inoculation both via gavage and feed. The parameters evaluated are weight, liver histopathology, liver and serum biochemistry, and bacteriological analyses of the caeca, crop, spleen, and liver and heart pool. At 21 and 28 days of age, the liver of the non-inoculated groups was significantly lighter as compared to the other treatments. Birds fed organic acids presented lower bacterial isolation rates in all organs tested. Birds inoculated in the crop and treated with organic acids presented lower E. coli CFU counts (P<0.05. Birds inoculated with Salmonella presented significant changes (P<0.05 in liver enzymes, as detected by serum biochemistry, and in liver histopathology. It was concluded that organic acids effectively controlled Salmonella Typhimurium and did not cause any liver damage.

Salmonella enterica serovars Typhimurium and Enteritidis causing mixed infections in febrile children in Mozambique

Directory of Open Access Journals (Sweden)

García V

2018-01-01

Full Text Available Vanesa García,1 Inácio Mandomando,2,3 Joaquim Ruiz,4 Silvia Herrera-León,5 Pedro L Alonso,3,4 M Rosario Rodicio1 1Departamento de Biología Funcional, Área de Microbiología, Universidad de Oviedo, Oviedo, Spain; 2Centro de Investigação em Saúde de Manhiça, 3Instituto Nacional de Saúde, Ministério da Saúde, Maputo, Mozambique; 4ISGlobal, Barcelona Centre for International Health Research, Hospital Clínic, Universitat de Barcelona, Barcelona, 5Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain Background and purpose: Invasive nontyphoidal salmonellosis, mostly caused by serovars Typhimurium and Enteritidis of Salmonella enterica, has emerged as a major public health problem in sub-Saharan Africa. The aim of this study was the clinical and microbiological characterization of nontyphoidal salmonellosis episodes affecting febrile children in Mozambique. Patients and methods: The clinical records of the patients were evaluated, and S. enterica isolates were characterized with regard to serovar, phage type, antimicrobial resistance (phenotype/responsible genes, plasmid content, pulsed-field gel electrophoresis, and multilocus sequence typing. Results: Fifteen S. Typhimurium and 21 S. Enteritidis isolates were recovered from blood samples of 25 children, the majority with underlying risk factors. With regard to phage typing, most isolates were either untypeable or reacted but did not conform, revealing that a number of previously unrecognized patterns are circulating in Mozambique. Most isolates were multidrug-resistant, with nearly all of the responsible genes located on derivatives of serovar-specific virulence plasmids. ST313 and ST11 were the predominant sequence types associated with S. Typhimurium and S. Enteritidis, respectively, and the uncommon ST1479 was also detected in S. Enteritidis. A distinct XbaI fragment of ~350 kb was associated with pulsed-field gel electrophoresis patterns of

Effect of microwave irradiation on Salmonella typhimurium cells

International Nuclear Information System (INIS)

Danilenko, I.I.; Mirutenko, V.I.; Sopil', A.V.; Koval'chuk, V.K.; Lyakhovchuk, N.N.; Popovich, G.G.; Bondarenko, V.I.

1985-01-01

It is shown that effect of electromagnetic energy of SHF-frequency, 8 mm wavelength and integral output power of 1MWt during 5.20 and 30 min results in negligible variations of Salmonella typhimurium cell ultrastructure. Increase of lipid peroxide amount determined according to malonic dialdehyde is observed in treated cells; it constitutes 10.23x10 -9 nm of malonic dialdehyde as compared to 3.20x10 -9 nm in control (untreated) cells

Identification of Salmonella Typhimurium-specific DNA aptamers developed using whole-cell SELEX and FACS analysis.

Science.gov (United States)

Moon, Jihea; Kim, Giyoung; Lee, Sangdae; Park, Saetbyeol

2013-11-01

Conventional methods for detection of infective organisms, such as Salmonella, are complicated and require multiple steps, and the need for rapid detection has increased. Biosensors show great potential for rapid detection of pathogens. In turn, aptamers have great potential for biosensor assay development, given their small size, ease of synthesis and labeling, lack of immunogenicity, a lower cost of production than antibodies, and high target specificity. In this study, ssDNA aptamers specific to Salmonella Typhimurium were obtained by a whole bacterium-based systematic evolution of ligands by exponential enrichment (SELEX) procedure and applied to probing S. Typhimurium. After 10 rounds of selection with S. Typhimurium as the target and Salmonella Enteritidis, Escherichia coli and Staphylococcus aureus as counter targets, the highly enriched oligonucleic acid pool was sorted using flow cytometry. In total, 12 aptamer candidates from different families were sequenced and grouped. Fluorescent analysis demonstrated that aptamer C4 had particularly high binding affinity and selectivity; this aptamer was then further characterized. © 2013 Elsevier B.V. All rights reserved.

Persistent Salmonella enterica serovar Typhimurium Infection Increases the Susceptibility of Mice to Develop Intestinal Inflammation

Directory of Open Access Journals (Sweden)

Bárbara M. Schultz

2018-05-01

Full Text Available Chronic intestinal inflammations are triggered by genetic and environmental components. However, it remains unclear how specific changes in the microbiota, host immunity, or pathogen exposure could promote the onset and exacerbation of these diseases. Here, we evaluated whether Salmonella enterica serovar Typhimurium (S. Typhimurium infection increases the susceptibility to develop intestinal inflammation in mice. Two mouse models were used to evaluate the impact of S. Typhimurium infection: the chemical induction of colitis by dextran sulfate sodium (DSS and interleukin (IL-10−/− mice, which develop spontaneous intestinal inflammation. We observed that S. Typhimurium infection makes DSS-treated and IL-10−/− mice more susceptible to develop intestinal inflammation. Importantly, this increased susceptibility is associated to the ability of S. Typhimurium to persist in liver and spleen of infected mice, which depends on the virulence proteins secreted by Salmonella Pathogenicity Island 2-encoded type three secretion system (TTSS-2. Although immunization with a live attenuated vaccine resulted in a moderate reduction of the IL-10−/− mice susceptibility to develop intestinal inflammation due to previous S. Typhimurium infection, it did not prevent bacterial persistence. Our results suggest that persistent S. Typhimurium infection may increase the susceptibility of mice to develop inflammation in the intestine, which could be associated with virulence proteins secreted by TTSS-2.

Antimicrobial resistance and typing of Salmonella isolated from street vended foods and associated environment.

Science.gov (United States)

Anukampa; Shagufta, Bi; Sivakumar, M; Kumar, Surender; Agarwal, Rajesh Kumar; Bhilegaonkar, Kiran Narayan; Kumar, Ashok; Dubal, Zunjar Baburao

2017-07-01

The present study was carried out to find out the occurrence and types of Salmonella present in street vended foods and associated environment, and their resistance pattern against various antibiotics. About 1075 street vended food and associated environment samples were processed for isolation and confirmation of different Salmonella spp. by targeting gene specific inv A gene and serotype specific Sdf I, Via B and Spy genes by PCR. Selected Salmonella isolates were screened for antibiotic resistance by using Baeur-Kirby disk diffusion test. Out of 1075 samples, only 31 (2.88%) isolates could be amplified the inv A gene of which 19 could be recovered from meat vendors; 8 from egg vendors while remaining 4 from milk vendors. Though, majority of Salmonella recovered from raw foods the ready-to-eat food like chicken gravy and rasmalai also showed its presence which pose a serious public health threat. Overall, 19, 6 and 1 isolates of S. Typhimurium, S. Enteritidis and S. Typhi could be detected by PCR while remaining 5 isolates could not be amplified suggesting other type of Salmonella. Selected Salmonella isolates were completely resistance to Oxacillin (100%) followed by Cefoxitin (30.43%) and Ampicillin (26.10%). Thus, it is observed that the street vended foods of animal origin and associated environment play an important role in transmission of food borne pathogens including Salmonella .

Prolonged restaurant-associated outbreak of multidrug-resistant Salmonella Typhimurium among patients from several European countries

DEFF Research Database (Denmark)

Ethelberg, S.; Lisby, M.; Torpdahl, M.

2004-01-01

This report concerns a prolonged restaurant-associated outbreak of infection caused by a multidrug-resistant (ASSuT) strain of Salmonella Typhimurium, phage-type U302, which took place during July and August 2003 and affected people from Denmark and neighbouring countries who had attended...... a specific restaurant. The outbreak comprised 67 laboratory-verified cases and ten probable cases; however, the actual number of patients was estimated to be more than 390. The outbreak strain was isolated from a buffet which was probably contaminated by an assistant chef who was found to excrete...

Identification by PCR of non-typhoidal Salmonella enterica serovars associated with invasive infections among febrile patients in Mali.

Directory of Open Access Journals (Sweden)

Sharon M Tennant

2010-03-01

Full Text Available In sub-Saharan Africa, non-typhoidal Salmonella (NTS are emerging as a prominent cause of invasive disease (bacteremia and focal infections such as meningitis in infants and young children. Importantly, including data from Mali, three serovars, Salmonella enterica serovar Typhimurium, Salmonella Enteritidis and Salmonella Dublin, account for the majority of non-typhoidal Salmonella isolated from these patients.We have extended a previously developed series of polymerase chain reactions (PCRs based on O serogrouping and H typing to identify Salmonella Typhimurium and variants (mostly I 4,[5],12:i:-, Salmonella Enteritidis and Salmonella Dublin. We also designed primers to detect Salmonella Stanleyville, a serovar found in West Africa. Another PCR was used to differentiate diphasic Salmonella Typhimurium and monophasic Salmonella Typhimurium from other O serogroup B, H:i serovars. We used these PCRs to blind-test 327 Salmonella serogroup B and D isolates that were obtained from the blood cultures of febrile patients in Bamako, Mali.We have shown that when used in conjunction with our previously described O-serogrouping PCR, our PCRs are 100% sensitive and specific in identifying Salmonella Typhimurium and variants, Salmonella Enteritidis, Salmonella Dublin and Salmonella Stanleyville. When we attempted to differentiate 171 Salmonella Typhimurium (I 4,[ 5],12:i:1,2 strains from 52 monophasic Salmonella Typhimurium (I 4,[5],12:i:- strains, we were able to correctly identify 170 of the Salmonella Typhimurium and 51 of the Salmonella I 4,[5],12:i:- strains.We have described a simple yet effective PCR method to support surveillance of the incidence of invasive disease caused by NTS in developing countries.

Risk factors associated with Salmonella enterica serovar typhimurium infection in Danish broiler flocks

DEFF Research Database (Denmark)

Skov, M. N.; Angen, Øystein; Chriel, M.

1999-01-01

A retrospective longitudinal study was conducted to identify risk factors associated with Salmonella enterica serovar typhimurium (S. typhimurium) infection in Danish broiler flocks. The data included all broiler flocks slaughtered in 1995, and the epidemiological unit was the individual broiler...... flock. The S. typhimurium status was determined by microbiological examination of 60 fresh fecal samples. This procedure should detect an infected flock with a probability above 95%, if the prevalence is above 5%, and given that the sensitivity of the test is 100%. Nineteen variables were selected...... for analysis. Five factors and an interaction term were found significant by multivariate logistic regression analysis. An increased risk for S, typhimurium infection was associated with two parent flocks, one confirmed infected and one suspected of being infected with S. typhimurium, with two...

Proteome analysis of serovars Typhimurium and Pullorum of Salmonella enterica subspecies I

Directory of Open Access Journals (Sweden)

Begum Shajna

2005-07-01

Full Text Available Abstract Background Salmonella enterica subspecies I includes several closely related serovars which differ in host ranges and ability to cause disease. The basis for the diversity in host range and pathogenic potential of the serovars is not well understood, and it is not known how host-restricted variants appeared and what factors were lost or acquired during adaptations to a specific environment. Differences apparent from the genomic data do not necessarily correspond to functional proteins and more importantly differential regulation of otherwise identical gene content may play a role in the diverse phenotypes of the serovars of Salmonella. Results In this study a comparative analysis of the cytosolic proteins of serovars Typhimurium and Pullorum was performed using two-dimensional gel electrophoresis and the proteins of interest were identified using mass spectrometry. An annotated reference map was created for serovar Typhimurium containing 233 entries, which included many metabolic enzymes, ribosomal proteins, chaperones and many other proteins characteristic for the growing cell. The comparative analysis of the two serovars revealed a high degree of variation amongst isolates obtained from different sources and, in some cases, the variation was greater between isolates of the same serovar than between isolates with different sero-specificity. However, several serovar-specific proteins, including intermediates in sulphate utilisation and cysteine synthesis, were also found despite the fact that the genes encoding those proteins are present in the genomes of both serovars. Conclusion Current microbial proteomics are generally based on the use of a single reference or type strain of a species. This study has shown the importance of incorporating a large number of strains of a species, as the diversity of the proteome in the microbial population appears to be significantly greater than expected. The characterisation of a diverse selection of

Modeling the survival of Salmonella Enteritidis and Salmonella Typhimurium during the fermentation of yogurt.

Science.gov (United States)

Savran, Derya; Pérez-Rodríguez, Fernando; Halkman, A Kadir

2018-03-01

The objective of this study was to evaluate the behavior of Salmonella Enteritidis and Salmonella Typhimurium, the two most important serovars of salmonellosis , during the fermentation of yogurt. The microorganisms were enumerated in milk throughout the fermentation process at three initial inoculum levels (3, 5 and 7 log CFU/mL). DMFit software was used in the fitting procedure of the data (IFR, Norwich, UK, Version 3.5). The data provided sigmoidal curves that were successfully displayed with the Baranyi model. The results showed that the initial inoculum level did not affect the growth for both pathogens; thus, the µ max values (maximum specific growth rate) did not significantly differ across all the contamination levels, ranging from 0.26 to 0.38 for S. Enteritidis and from 0.50 to 0.56 log CFU/g/h for S. Typhimurium ( P > 0.05). However, the µ max values significantly differed between the two serovars ( P fermentation process of milk even at a low contamination level. In addition, the models presented in this study can be used in quantitative risk assessment studies to estimate the threat to consumers.

Genetic Transfer of Salmonella typhimurium and Escherichia coli Lipopolysaccharide Antigens to Escherichia coli K-12

Science.gov (United States)

Jones, Randall T.; Koeltzow, Donald E.; Stocker, B. A. D.

1972-01-01

Escherichia coli K-12 ϰ971 was crossed with a smooth Salmonella typhimurium donor, HfrK6, which transfers early the ilv-linked rfa region determining lipopolysaccharide (LPS) core structure. Two ilv+ hybrids differing in their response to the LPS-specific phages FO and C21 were then crossed with S. typhimurium HfrK9, which transfers early the rfb gene cluster determining O repeat unit structure. Most recombinants selected for his+ (near rfb) were agglutinated by Salmonella factor 4 antiserum. Transfer of an F′ factor (FS400) carrying the rfb–his region of S. typhimurium to the same two ilv+ hybrids gave similar results. LPS extracted from two ilv+,his+, factor 4-positive hybrids contained abequose, the immunodominant sugar for factor 4 specificity. By contrast, his+ hybrids obtained from ϰ971 itself by similar HfrK9 and F′FS400 crosses were not agglutinated by factor 4 antiserum, indicating that the parental E. coli ϰ971 does not have the capacity to attach Salmonella O repeat units to its LPS core. It is concluded that the Salmonella rfb genes are expressed only in E. coli ϰ971 hybrids which have also acquired ilv-linked genes (presumably rfa genes affecting core structure or O-translocase ability, or both) from a S. typhimurium donor. When E. coli ϰ971 was crossed with a smooth E. coli donor, Hfr59, of serotype O8, which transfers his early, most his+ recombinants were agglutinated by E. coli O8 antiserum and lysed by the O8-specific phage, Ω8. This suggests that, although the parental E. coli K-12 strain ϰ971 cannot attach Salmonella-specific repeat units to its LPS core, it does have the capacity to attach E. coli O8-specific repeat units. PMID:4559827

Effect of coffee filtrate, methylglyoxal, glyoxal, and caffeine on Salmonella typhimurium and S. enteritidis survival in ground chicken breasts.

Science.gov (United States)

Maletta, Anne B; Were, Lilian M

2012-02-01

The antimicrobial effect of roasted coffee filtrate (CF) and dicarbonyls on Salmonella Typhimurium and Salmonella Enteritidis in raw ground chicken breast meat (GCB) was investigated. Coffee was brewed and filtered before addition to GCB. Coffee filtrate with and without added caffeine, methylglyoxal, and/or glyoxal was added to GCB and then inoculated with Salmonella Typhimurium and Salmonella Enteritidis. Ground chicken samples were stomached with peptone water at days 1, 3, 5, and 7, plated on XLD agar with a TSA overlay, and Salmonella survivors were enumerated. CF alone gave less than a 1 Log reduction in all runs compared to control GCB with no treatment. Methylglyoxal (2.28 mg/g GCB) had the greatest antimicrobial effect against Salmonella Typhimurium and Salmonella Enteritidis in GCB with average Log reductions of 2.27 to 3.23, respectively, over the 7 d duration of the experiment compared to control GCB with no treatment. A 1 Log reduction was observed in GCB with CF, 0.93 mg glyoxal, and 1 mg caffeine/g chicken compared to the control and GCB with only CF. Heat-produced coffee compounds could potentially reduce Salmonella in retail ground chicken and chicken products. © 2011 Institute of Food Technologists®

Virulence characterisation of Salmonella enterica isolates of differing antimicrobial resistance recovered from UK livestock and imported meat samples.

Directory of Open Access Journals (Sweden)

Roderick eCard

2016-05-01

Full Text Available Salmonella enterica is a foodborne zoonotic pathogen of significant public health concern. We have characterised the virulence and antimicrobial resistance gene content of 95 Salmonella isolates from 11 serovars by DNA microarray recovered from UK livestock or imported meat. Genes encoding resistance to sulphonamides (sul1, sul2, tetracycline (tet(A, tet(B, streptomycin (strA, strB, aminoglycoside (aadA1, aadA2, beta-lactam (blaTEM, and trimethoprim (dfrA17 were common. Virulence gene content differed between serovars; S. Typhimurium formed two subclades based on virulence plasmid presence. Thirteen isolates were selected by their virulence profile for pathotyping using the Galleria mellonella pathogenesis model. Infection with a chicken invasive S. Enteritidis or S. Gallinarum isolate, a multidrug resistant S. Kentucky, or a S. Typhimurium DT104 isolate resulted in high mortality of the larvae; notably presence of the virulence plasmid in S. Typhimurium was not associated with increased larvae mortality. Histopathological examination showed that infection caused severe damage to the Galleria gut structure. Enumeration of intracellular bacteria in the larvae 24 hours post-infection showed increases of up to 7 log above the initial inoculum and transmission electron microscopy (TEM showed bacterial replication in the haemolymph. TEM also revealed the presence of vacuoles containing bacteria in the haemocytes, similar to Salmonella containing vacuoles observed in mammalian macrophages; although there was no evidence from our work of bacterial replication within vacuoles. This work shows that microarrays can be used for rapid virulence genotyping of S. enterica and that the Galleria animal model replicates some aspects of Salmonella infection in mammals. These procedures can be used to help inform on the pathogenicity of isolates that may be antibiotic resistant and have scope to aid the assessment of their potential public and animal health risk.

Decrease of Salmonella typhimurium in skim milk and egg by heat and ultrasonic wave treatment

International Nuclear Information System (INIS)

Wrigley, D.M.; Llorca, N.G.

1992-01-01

Ultrasonic waves induce cavitation which is lethal for many bacteria. When Salmonella typhimurium was suspended in skim milk or brain heart infusion broth and placed in an ultrasonicating water bath, the number of bacteria decreased by 2 to 3 log CFU in a time dependent manner. The killing by ultrasonic waves was enhanced if the menstruum was simultaneously maintained at 50 degrees C. Ultrasonic reduction in S. typhimurium numbers in liquid whole egg ranged from 1-3 log CFU at 50 degrees C. The results indicate that indirect ultrasonic wave treatment is effective in killing Salmonella in some foods

In Vitro Assembly of the Outer Core of the Lipopolysaccharide from Escherichia coli K-12 and Salmonella typhimurium

Science.gov (United States)

2015-01-01

There are five distinct core structures in the lipopolysaccharides of Escherichia coli and at least two in Salmonella isolates, which vary principally in the outer core oligosaccharide. Six outer core glycosyltransferases, E. coli K-12 WaaG, WaaB, and WaaO and Salmonella typhimurium WaaI, WaaJ, and WaaK, were cloned, overexpressed, and purified. A novel substrate for WaaG was isolated from ΔwaaG E. coli overexpressing the lipid A phosphatase lpxE and the lipid A late acyltransferase lpxM. The action of lpxE and lpxM in the ΔwaaG background yielded heptose2-1-dephospho Kdo2-lipid A, a 1-dephosphorylated hexa-acylated lipid A with the inner core sugars that is easily isolated by organic extraction. Using this structurally defined acceptor and commercially available sugar nucleotides, each outer core glycosyltransferases was assayed in vitro. We show that WaaG and WaaB add a glucose and galactose sequentially to heptose2-1-dephospho Kdo2-lipid A. E. coli K-12 WaaO and S. typhimurium WaaI add a galactose to the WaaG/WaaB product but can also add a galactose to the WaaG product directly without the branched core sugar added by WaaB. Both WaaI and WaaO require divalent metal ions for optimal activity; however, WaaO, unlike WaaI, can add several glucose residues to its lipid acceptor. Using the product of WaaG, WaaB, and WaaI, we show that S. typhimurium WaaJ and WaaK transfer a glucose and N-acetylglucosamine, respectively, to yield the full outer core. This is the first demonstration of the in vitro assembly of the outer core of the lipopolysaccharide using defined lipid A-oligosaccharide acceptors and sugar donors. PMID:24479701

Characterisation of integrons and antibiotic resistance genes in Danish multiresistant Salmonella enterica Typhimurium DT104

DEFF Research Database (Denmark)

Sandvang, Dorthe; Aarestrup, Frank Møller; Jensen, Lars Bogø

1997-01-01

The presence and genetic content of integrons was investigated in eight Salmonella enterica Typhimurium DT104 isolates from different pig herds in Denmark. Two different integrons were identified using PCR and sequencing. Each of the integrons carried a single resistance cassette in addition...... to the sul1 and qacE Delta 1 genes characteristic of integrons. The first integron encoded the ant (3 ")-Ia gene that specified resistance to spectinomycin and streptomycin. The second contained the pse-l beta-lactamase gene. All the multiresistant strains contained both integrons. The presence of these two...... integrons did not account for the total phenotypic resistance of all the isolates and does not exclude the presence of other mobile DNA elements....

Characterisation of integrons and antibiotic resistance genes in Danish multiresistant Salmonella enterica Typhimurium DT104

DEFF Research Database (Denmark)

Sandvang, Dorthe; Aarestrup, Frank Møller; Jensen, Lars Bogø

1998-01-01

The presence and genetic content of integrons was investigated in eight Salmonella enteritica Typhimurium DT104 isolates from different pig herds in Denmark. Two different integrons were identified using PCR and sequencing. Each of the integrons carried a single resistance cassette in addition...... to the sul1 and qacE Delta 1 genes characteristic of integrons. The first integron encoded the ant (3")-Ia gene that specified resistance to spectinomycin and streptomycin. The second contained the pse-1 beta-lactamase gene. All the multiresistant strains contained both integrons. The presence of these two...... integrons did not account for the total phenotypic resistance of all the isolates and does not exclude the presence of other mobile DNA elements....

Antimicrobial Resistance of Enteric Salmonella in Bangui, Central African Republic

Directory of Open Access Journals (Sweden)

Christian Diamant Mossoro-Kpinde

2015-01-01

Full Text Available Introduction. The number of Salmonella isolated from clinical samples that are resistant to multiple antibiotics has increased worldwide. The aim of this study was to determine the prevalence of resistant Salmonella enterica isolated in Bangui. Methods. All enteric Salmonella strains isolated from patients in 2008 were identified and serotyped, and the phenotypes of resistance were determined by using the disk diffusion method. Nine resistance-associated genes, blaTEM, blaOXA, blaSHV, tetA, aadA1, catA1, dhfrA1, sul I, and sul II, were sought by genic amplification in seven S.e. Typhimurium strains. Results. The 94 strains isolated consisted of 47 S.e. Typhimurium (50%, 21 S.e. Stanleyville (22%, 18 S.e. Enteritidis (19%, 4 S.e. Dublin (4%, 4 S.e. Hadar (4%, and 1 S.e. Papuana (1%. Twenty-five (28% were multiresistant, including 20 of the Typhimurium serovar (80%. Two main phenotypes of resistance were found: four antibiotics (56% and to five antibiotics (40%. One S.e. Typhimurium isolate produced an extended-spectrum β-lactamase (ESBL. Only seven strains of S.e. Typhimurium could be amplified genically. Only phenotypic resistance to tetracycline and aminosides was found. Conclusion. S. Typhimurium is the predominant serovar of enteric S. enterica and is the most widely resistant. The search for resistance genes showed heterogeneity of the circulating strains.

Salmonella Typhimurium type III secretion effectors stimulate innate immune responses in cultured epithelial cells.

Directory of Open Access Journals (Sweden)

Vincent M Bruno

2009-08-01

Full Text Available Recognition of conserved bacterial products by innate immune receptors leads to inflammatory responses that control pathogen spread but that can also result in pathology. Intestinal epithelial cells are exposed to bacterial products and therefore must prevent signaling through innate immune receptors to avoid pathology. However, enteric pathogens are able to stimulate intestinal inflammation. We show here that the enteric pathogen Salmonella Typhimurium can stimulate innate immune responses in cultured epithelial cells by mechanisms that do not involve receptors of the innate immune system. Instead, S. Typhimurium stimulates these responses by delivering through its type III secretion system the bacterial effector proteins SopE, SopE2, and SopB, which in a redundant fashion stimulate Rho-family GTPases leading to the activation of mitogen-activated protein (MAP kinase and NF-kappaB signaling. These observations have implications for the understanding of the mechanisms by which Salmonella Typhimurium induces intestinal inflammation as well as other intestinal inflammatory pathologies.

Detection of Salmonella Typhimurium on Spinach Using Phage-Based Magnetoelastic Biosensors

Directory of Open Access Journals (Sweden)

Fengen Wang

2017-02-01

Full Text Available Phage-based magnetoelastic (ME biosensors have been studied as an in-situ, real-time, wireless, direct detection method of foodborne pathogens in recent years. This paper investigates an ME biosensor method for the detection of Salmonella Typhimurium on fresh spinach leaves. A procedure to obtain a concentrated suspension of Salmonella from contaminated spinach leaves is described that is based on methods outlined in the U.S. FDA Bacteriological Analytical Manual for the detection of Salmonella on leafy green vegetables. The effects of an alternative pre-enrichment broth (LB broth vs. lactose broth, incubation time on the detection performance and negative control were investigated. In addition, different blocking agents (BSA, Casein, and Superblock were evaluated to minimize the effect of nonspecific binding. None of the blocking agents was found to be superior to the others, or even better than none. Unblocked ME biosensors were placed directly in a concentrated suspension and allowed to bind with Salmonella cells for 30 min before measuring the resonant frequency using a surface-scanning coil detector. It was found that 7 h incubation at 37 °C in LB broth was necessary to detect an initial spike of 100 cfu/25 g S. Typhimurium on spinach leaves with a confidence level of difference greater than 95% (p < 0.05. Thus, the ME biosensor method, on both partly and fully detection, was demonstrated to be a robust and competitive method for foodborne pathogens on fresh products.

Persistence of Salmonella Typhimurium LT2 in Soil Enhanced after Growth in Lettuce Medium

Directory of Open Access Journals (Sweden)

Kornelia Smalla

2017-04-01

Full Text Available The persistence of Salmonella in the environment is influenced by a multitude of biotic and abiotic factors. In addition, its persistence can be influenced by preadaptation before the introduction into the environment. In order to study how preadaptation changes the survival of Salmonella in soil and therefore its potential to colonize the phytosphere, we developed a new medium based on lettuce material [lettuce medium (LM]. Salmonella enterica serovar Typhimurium strain LT2 was used as a model for Salmonella in this study. LT2 was inoculated into soil microcosms after pregrowth in Luria Bertani (LB broth or in LM. Survival of LT2 in soil was monitored over 56 days by plate counts and quantification of the Typhimurium-specific gene STM4497 using qPCR in total community DNA for which primers and TaqMan probe were designed in this study. Significantly enhanced persistence was observed for LT2 pregrown in LM compared to LT2 pregrown in LB, indicating a preadaptation effect. Surprisingly, no improved survival could be observed for S. Typhimurium strain 14028s and S. enterica serovar Senftenberg after pregrowth on LM. This indicates a high strain specificity of preadaptation. Results from previous studies suggested that biofilm formation could enhance the survival of human pathogens in various environments and might contribute to enhanced survival on plants. In vitro biofilm assays with several Salmonella strains revealed a strain-specific effect of LM on the biofilm formation. While LM significantly improved the biofilm formation of S. Senftenberg, the biofilm formation of LT2 was better in LB. This indicates that the better survival of LM-pregrown LT2 in soil was not linked to an improved ability to form biofilms but was likely due to other factors. Most importantly, this study showed that the medium used to pregrow Salmonella can influence its survival in soil and its biofilm formation which might influence the fate of Salmonella in soil.

Antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. isolated from chicken samples.

Science.gov (United States)

Er, Buket; Demirhan, Burak; Onurdag, Fatma Kaynak; Ozgacar, Selda Özgen; Oktem, Aysel Bayhan

2014-03-01

Salmonella spp. are widespread foodborne pathogens that contaminate egg and poultry meats. Attachment, colonization, as well as biofilm formation capacity of Salmonella spp. on food and contact surfaces of food may cause continuous contamination. Biofilm may play a crucial role in the survival of salmonellae under unfavorable environmental conditions, such as in animal slaughterhouses and processing plants. This could serve as a reservoir compromising food safety and human health. Addition of antimicrobial preservatives extends shelf lives of food products, but even when products are supplemented with adequate amounts of preservatives, it is not always possible to inhibit the microorganisms in a biofilm community. In this study, our aims were i) to determine the minimum inhibitory concentrations (MIC) and minimum biofilm inhibitory concentrations (MBIC) of selected preservatives against planktonic and biofilm forms of Salmonella spp. isolated from chicken samples and Salmonella Typhimurium SL1344 standard strain, ii) to show the differences in the susceptibility patterns of same strains versus the planktonic and biofilm forms to the same preservative agent, and iii) to determine and compare antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. For this purpose, Salmonella Typhimurium SL1344 standard strain and 4 Salmonella spp. strains isolated from chicken samples were used. Investigation of antimicrobial and antibiofilm effects of selected food preservatives against Salmonella spp. was done according to Clinical and Laboratory Standards Institute M100-S18 guidelines and BioTimer assay, respectively. As preservative agents, pure ciprofloxacin, sodium nitrite, potassium sorbate, sodium benzoate, methyl paraben, and propyl paraben were selected. As a result, it was determined that MBIC values are greater than the MIC values of the preservatives. This result verified the resistance seen in a biofilm community to food

Some putative prebiotics increase the severity of Salmonella enterica serovar Typhimurium infection in mice

Directory of Open Access Journals (Sweden)

Lahtinen Sampo

2009-01-01

Full Text Available Abstract Background Prebiotics are non-digestible food ingredients believed to beneficially affect host health by selectively stimulating the growth of the beneficial bacteria residing in the gut. Such beneficial bacteria have been reported to protect against pathogenic infections. However, contradicting results on prevention of Salmonella infections with prebiotics have been published. The aim of the present study was to examine whether S. Typhimurium SL1344 infection in mice could be prevented by administration of dietary carbohydrates with different structures and digestibility profiles. BALB/c mice were fed a diet containing 10% of either of the following carbohydrates: inulin, fructo-oligosaccharide, xylo-oligosaccharide, galacto-oligosaccharide, apple pectin, polydextrose or beta-glucan for three weeks prior to oral Salmonella challenge (107 CFU and compared to mice fed a cornstarch-based control diet. Results The mice fed with diets containing fructo-oligosaccharide (FOS or xylo-oligosaccharide (XOS had significantly higher (P < 0.01 and P < 0.05 numbers of S. Typhimurium SL1344 in liver, spleen and mesenteric lymph nodes when compared to the mice fed with the cornstarch-based control diet. Significantly increased amounts (P < 0.01 of Salmonella were detected in ileal and fecal contents of mice fed with diets supplemented with apple pectin, however these mice did not show significantly higher numbers of S. Typhimyrium in liver, spleen and lymph nodes than animals from the control group (P < 0.20. The acute-phase protein haptoglobin was a good marker for translocation of S. Typhimurium in mice. In accordance with the increased counts of Salmonella in the organs, serum concentrations of haptoglobin were significantly increased in the mice fed with FOS or XOS (P < 0.001. Caecum weight was increased in the mice fed with FOS (P < 0.01, XOS (P < 0.01, or polydextrose (P < 0.001, and caecal pH was reduced in the mice fed with polydextrose (P < 0

Affinity-Selected Filamentous Bacteriophage as a Probe for Acoustic Wave Biodetectors of Salmonella typhimurium

National Research Council Canada - National Science Library

Olsen, Eric V; Sorokulova, Iryna B; Petrenko, Valery A; Chen, I-Hsuan; Barbaree, James M; Vodyanoy, Vitaly J

2005-01-01

Proof-in-concept biosensors were prepared for the rapid detection of Salmonella typhimurium in solution, based on affinity-selected filamentous phage prepared as probes physically adsorbed to piezoelectric transducers...

Analysis of the ArcA regulon in anaerobically grown Salmonella enterica sv. Typhimurium

Directory of Open Access Journals (Sweden)

Porwollik Steffen

2011-03-01

Full Text Available Abstract Background Salmonella enterica serovar Typhimurium (S. Typhimurium is a Gram-negative pathogen that must successfully adapt to the broad fluctuations in the concentration of dissolved dioxygen encountered in the host. In Escherichia coli, ArcA (Aerobic Respiratory Control helps the cells to sense and respond to the presence of dioxygen. The global role of ArcA in E. coli is well characterized; however, little is known about its role in anaerobically grown S. Typhimurium. Results We compared the transcriptional profiles of the virulent wild-type (WT strain (ATCC 14028s and its isogenic arcA mutant grown under anaerobic conditions. We found that ArcA directly or indirectly regulates 392 genes (8.5% of the genome; of these, 138 genes are poorly characterized. Regulation by ArcA in S. Typhimurium is similar, but distinct from that in E. coli. Thus, genes/operons involved in core metabolic pathways (e.g., succinyl-CoA, fatty acid degradation, cytochrome oxidase complexes, flagellar biosynthesis, motility, and chemotaxis were regulated similarly in the two organisms. However, genes/operons present in both organisms, but regulated differently by ArcA in S. Typhimurium included those coding for ethanolamine utilization, lactate transport and metabolism, and succinate dehydrogenases. Salmonella-specific genes/operons regulated by ArcA included those required for propanediol utilization, flagellar genes (mcpAC, cheV, Gifsy-1 prophage genes, and three SPI-3 genes (mgtBC, slsA, STM3784. In agreement with our microarray data, the arcA mutant was non-motile, lacked flagella, and was as virulent in mice as the WT. Additionally, we identified a set of 120 genes whose regulation was shared with the anaerobic redox regulator, Fnr. Conclusion(s We have identified the ArcA regulon in anaerobically grown S. Typhimurium. Our results demonstrated that in S. Typhimurium, ArcA serves as a transcriptional regulator coordinating cellular metabolism, flagella

Genetic diversity and antimicrobial resistance of Campylobacter and Salmonella strains isolated from decoys and raptors.

Science.gov (United States)

Jurado-Tarifa, E; Torralbo, A; Borge, C; Cerdà-Cuéllar, M; Ayats, T; Carbonero, A; García-Bocanegra, I

2016-10-01

Infections caused by thermotolerant Campylobacter spp. and Salmonella spp. are the leading causes of human gastroenteritis worldwide. Wild birds can act as reservoirs of both pathogens. A survey was carried out to determine the prevalence, genetic diversity and antimicrobial resistance of thermotolerant Campylobacter and Salmonella in waterfowl used as decoys and wild raptors in Andalusia (Southern Spain). The overall prevalence detected for Campylobacter was 5.9% (18/306; CI95%: 3.25-8.52) in decoys and 2.3% (9/387; CI95%: 0.82-3.83) in wild raptors. Isolates were identified as C. jejuni, C. coli and C. lari in both bird groups. Salmonella was isolated in 3.3% (10/306; CI95%: 2.3-4.3) and 4.6% (18/394; CI95%: 3.5-5.6) of the decoys and raptors, respectively. Salmonella Enteritidis and Typhimurium were the most frequently identified serovars, although Salmonella serovars Anatum, Bredeney, London and Mikawasima were also isolated. Pulsed-field gel electrophoresis analysis of isolates showed higher genetic diversity within Campylobacter species compared to Salmonella serovars. Campylobacter isolates showed resistance to gentamicin, ciprofloxacin and tetracycline, while resistance to erythromycin and tetracycline was found in Salmonella isolates. The results indicate that both decoys and raptors can act as natural carriers of Campylobacter and Salmonella in Spain, which may have important implications for public and animal health. Copyright © 2016 Elsevier Ltd. All rights reserved.

A questionnaire-based, retrospective field study of persistence of Salmonella Enteritidis and Salmonella Typhimurium in Danish broiler houses

DEFF Research Database (Denmark)

Gradel, K.O.; Rattenborg, Erik

2003-01-01

A questionnaire-based, retrospective field study was conducted in 78 Danish broiler houses (analytical units) on 42 farms. In spring 1997, all these broiler houses had been infected with Salmonella Enteritidis, phage type 8, and/or Salmonella Typhimurium, definitive-type 66, by day-old chicks del...... soap and water for washing hands in the anteroom, hygiene barriers when removing dead broilers, gravel alongside the broiler house, systematic checks of indoor rodent-bait depots, and combined surface and pulse-fogging disinfection....

Global Genomic Epidemiology of Salmonella enterica Serovar Typhimurium DT104

DEFF Research Database (Denmark)

Leekitcharoenphon, Pimlapas; Hendriksen, Rene S.; Le Hello, Simon

2016-01-01

It has been 30 years since the initial emergence and subsequent rapid global spread of multidrug-resistant Salmonella enterica serovar Typhimurium DT104 (MDR DT104). Nonetheless, its origin and transmission route have never been revealed. We used whole-genome sequencing (WGS) and temporally struc...

Primary structure and mapping of the hupA gene of Salmonella typhimurium.

OpenAIRE

Higgins, N P; Hillyard, D

1988-01-01

In bacteria, the complex nucleoid structure is folded and maintained by negative superhelical tension and a set of type II DNA-binding proteins, also called histonelike proteins. The most abundant type II DNA-binding protein is HU. Southern blot analysis showed that Salmonella typhimurium contained two HU genes that corresponded to Escherichia coli genes hupA (encoding HU-2 protein) and hupB (encoding HU-1). Salmonella hupA was cloned, and the nucleotide sequence of the gene was determined. C...

Breast abscess due to Salmonella Typhimurium in a patient with rheumatoid arthritis: a case report.

Science.gov (United States)

Baran, Irmak; Aksu, Neriman; Aksoy, Altan

2016-07-22

This is the first report of breast abscess due to Salmonella enterica serotype Typhimurium. Staphylococcus aureus is known as the most common cause of breast abscess. Salmonella spp. may occasionally form localized abscesses after dissemination to various organ systems following a bacteraemia. But breast abscess related to Salmonella spp is a very rare complication. A 43-year-old female patient referred to our hospital with a lump, fever and mild pain in her breast. The patient was not pregnant or lactating at that time. She had a history of rheumatoid arthritis for 5 years and was under immunosuppressive therapy. Ultrasonography of the breast revealed an abscess. The abscess was drained and sent for culture to medical microbiology laboratory. The microorganism was identified as Salmonella enterica serotype Typhimurium and found to be sensitive to all antibiotics tested. The patient was cured after surgical debridement and antibiotic therapy. The abscess did not recur again. This case is presented to draw attention to non-typhoidal Salmonella as rare causes of breast abscess and submission of specimens to the microbiology laboratory for accurate diagnosis and treatment especially in patients with underlying immunosuppressive diseases.

Studies on the interaction between Salmonella enterica ser. Typhimurium and intestinal helminths in pigs

DEFF Research Database (Denmark)

Steenhard, N.R.; Roepstorff, A.; Baggesen, Dorte Lau

2006-01-01

Concomitant infections with helminths and bacteria may affect the course and the resulting disease outcome of the individual infections. Salmonella, Oesophagostomum, Trichuris and Ascaris coexist naturally in pig herds in Denmark, and possible interactions were studied. Pigs in one experiment were...... was not demonstrated in either experiment. The helminth effect on the pigs was modest and may explain the lack of influence on the Salmonella infection. A previous experiment with a larger Oesophagostomum infection level resulted in enhancement of the S. Typhimurium infection. A dose dependency of the interaction...... is therefore suggested. However, the relatively high worm burdens in the present study suggest that infection with these common pig helminths does generally not influence the course of concurrent S. Typhimurium infections under natural conditions....

Inactivation of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella typhimurium with compounds available in households.

Science.gov (United States)

Yang, Hua; Kendall, Patricia A; Medeiros, Lydia; Sofos, John N

2009-06-01

Solutions of selected household products were tested for their effectiveness against Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Typhimurium. Hydrogen peroxide (1.5 and 3%), vinegar (2.5 and 5% acetic acid), baking soda (11, 33, and 50% sodium bicarbonate), household bleach (0.0314, 0.0933, and 0.670% sodium hypochlorite), 5% acetic acid (prepared from glacial acetic acid), and 5% citric acid solutions were tested against the three pathogens individually (five-strain composites of each, 10(8) CFU/ml) by using a modified AOAC International suspension test at initial temperatures of 25 and 55degrees C for 1 and 10 min. All bleach solutions (pH 8.36 to 10.14) produced a >5-log reduction of all pathogens tested after 1 min at 25 degrees C, whereas all baking soda solutions (pH 7.32 to 7.55) were ineffective (5-log reduction of both Salmonella Typhimurium and E. coli O157:H7, whereas undiluted vinegar (pH 2.58) had a similar effect only against Salmonella Typhimurium. Compared with 1 min at 25 degrees C, greater reductions of L. monocytogenes (P 3% hydrogen peroxide > undiluted vinegar and 5% acetic acid > 5% citric acid > baking soda (50% sodium bicarbonate). The sensitivity of the tested pathogens to all tested household compounds followed the sequence of Salmonella Typhimurium > E. coli O157: H7 > L. monocytogenes.

Antimicrobial susceptibility and internalization of Salmonella Typhimurium in vacuum-tumbled marinated beef products.

Science.gov (United States)

Pokharel, S; Brooks, J C; Martin, J N; Brashears, M M

2016-12-01

As the incidence of multidrug resistance (MDR) Salmonella enterica serotype Typhimurium is increasing, data regarding the antimicrobial interventions and pathogen internalization in marinated meat products are important. This study evaluated the antimicrobial intervention and internalization of Salm. Typhimurium in marinated beef sirloin steaks. Beef bottom sirloin flaps (IMPS #185A; USDA Select) inoculated (10 8  log 10  CFU ml -1 ) with Salm. Typhimurium were sprayed (lactic acid (4%) and buffered vinegar (2%)) prior to vacuum-tumbled marination (0·35% sodium chloride and 0·45% sodium tripolyphosphate) for 30 min. Pathogen presence after antimicrobial spray, vacuum-tumbled marination, and translocation was determined by direct plating on Xylose Lysine Deoxycholate (XLD) agar with tryptic soy agar (TSA) overlay. The data imply varied internalization and antimicrobial susceptibility pattern of Salm. Typhimurium in marinated meat. Lactic acid (4%) spray (P internalization and potential survival of Salmonella spp. in marinated beef products is a major concern. These results highlight the internalization of pathogens in vacuum-tumbled meat products and emphasize the importance of considering these products as nonintact. Similarly, these data confirm the efficacy and utility of interventions prior to vacuum-tumbled marination. Further research is needed to identify additional strategies to mitigate internalization and translocation of pathogens into vacuum-marinated meat products. © 2016 The Society for Applied Microbiology.

The transcriptional programme of Salmonella enterica serovar Typhimurium reveals a key role for tryptophan metabolism in biofilms.

LENUS (Irish Health Repository)

Hamilton, Shea

2009-12-11

Abstract Background Biofilm formation enhances the capacity of pathogenic Salmonella bacteria to survive stresses that are commonly encountered within food processing and during host infection. The persistence of Salmonella within the food chain has become a major health concern, as biofilms can serve as a reservoir for the contamination of food products. While the molecular mechanisms required for the survival of bacteria on surfaces are not fully understood, transcriptional studies of other bacteria have demonstrated that biofilm growth triggers the expression of specific sets of genes, compared with planktonic cells. Until now, most gene expression studies of Salmonella have focused on the effect of infection-relevant stressors on virulence or the comparison of mutant and wild-type bacteria. However little is known about the physiological responses taking place inside a Salmonella biofilm. Results We have determined the transcriptomic and proteomic profiles of biofilms of Salmonella enterica serovar Typhimurium. We discovered that 124 detectable proteins were differentially expressed in the biofilm compared with planktonic cells, and that 10% of the S. Typhimurium genome (433 genes) showed a 2-fold or more change in the biofilm compared with planktonic cells. The genes that were significantly up-regulated implicated certain cellular processes in biofilm development including amino acid metabolism, cell motility, global regulation and tolerance to stress. We found that the most highly down-regulated genes in the biofilm were located on Salmonella Pathogenicity Island 2 (SPI2), and that a functional SPI2 secretion system regulator (ssrA) was required for S. Typhimurium biofilm formation. We identified STM0341 as a gene of unknown function that was needed for biofilm growth. Genes involved in tryptophan (trp) biosynthesis and transport were up-regulated in the biofilm. Deletion of trpE led to decreased bacterial attachment and this biofilm defect was restored by

An incomplete TCA cycle increases survival of Salmonella Typhimurium during infection of resting and activated murine macrophages.

Science.gov (United States)

Bowden, Steven D; Ramachandran, Vinoy K; Knudsen, Gitte M; Hinton, Jay C D; Thompson, Arthur

2010-11-08

In comparison to the comprehensive analyses performed on virulence gene expression, regulation and action, the intracellular metabolism of Salmonella during infection is a relatively under-studied area. We investigated the role of the tricarboxylic acid (TCA) cycle in the intracellular replication of Salmonella Typhimurium in resting and activated macrophages, epithelial cells, and during infection of mice. We constructed deletion mutations of 5 TCA cycle genes in S. Typhimurium including gltA, mdh, sdhCDAB, sucAB, and sucCD. We found that the mutants exhibited increased net intracellular replication in resting and activated murine macrophages compared to the wild-type. In contrast, an epithelial cell infection model showed that the S. Typhimurium ΔsucCD and ΔgltA strains had reduced net intracellular replication compared to the wild-type. The glyoxylate shunt was not responsible for the net increased replication of the TCA cycle mutants within resting macrophages. We also confirmed that, in a murine infection model, the S. Typhimurium ΔsucAB and ΔsucCD strains are attenuated for virulence. Our results suggest that disruption of the TCA cycle increases the ability of S. Typhimurium to survive within resting and activated murine macrophages. In contrast, epithelial cells are non-phagocytic cells and unlike macrophages cannot mount an oxidative and nitrosative defence response against pathogens; our results show that in HeLa cells the S. Typhimurium TCA cycle mutant strains show reduced or no change in intracellular levels compared to the wild-type. The attenuation of the S. Typhimurium ΔsucAB and ΔsucCD mutants in mice, compared to their increased net intracellular replication in resting and activated macrophages suggest that Salmonella may encounter environments within the host where a complete TCA cycle is advantageous.

Rapid detection of Salmonella typhimurium on fresh spinach leaves using phage-immobilized magnetoelastic biosensors

Science.gov (United States)

Horikawa, Shin; Li, Suiqiong; Chai, Yating; Park, Mi-Kyung; Shen, Wen; Barbaree, James M.; Vodyanoy, Vitaly J.; Chin, Bryan A.

2011-06-01

This paper presents an investigation into the use of magnetoelastic biosensors for the rapid detection of Salmonella typhimurium on fresh spinach leaves. The biosensors used in this investigation were comprised of a strip-shaped, goldcoated sensor platform (2 mm-long) diced from a ferromagnetic, amorphous alloy and a filamentous fd-tet phage which specifically binds with S. typhimurium. After surface blocking with bovine serum albumin, these biosensors were, without any preceding sample preparation, directly placed on wet spinach leaves inoculated with various concentrations of S. typhimurium. Upon contact with cells, the phage binds S. typhimurium to the sensor thereby increasing the total mass of the sensor. This change in mass causes a corresponding decrease in the sensor's resonant frequency. After 25 min, the sensors were collected from the leaf surface and measurements of the resonant frequency were performed immediately. The total assay time was less than 30 min. The frequency changes for measurement sensors (i.e., phageimmobilized) were found to be statistically different from those for control sensors (sensors without phage), down to 5 × 106 cells/ml. The detection limit may be improved by using smaller, micron-sized sensors that will have a higher probability of contacting Salmonella on the rough surfaces of spinach leaves.

Effect of the irradiation on Salmonella enteretidis var. typhimurium with gamma rays from 60Co

International Nuclear Information System (INIS)

Maliska, C.; Norberg, A.N.; Norberg, J.B.M.; Santos Souza, R.C. dos; Silva Tavares, P.R. da

1988-01-01

The use of ionizinf radiation to the destruction of microrganisms responsible for food deterioration, and productive of feeding toxinfections constitute their usefulness for actually peaceful goals of nuclear energy. The feeding toxinfections are, among us, produced in their most part by Salmonella enteritidis var. typhimurim. One hundred nineteen samples of milk containing about 150.000 bacteria per ml, by means doses ranging from 100 to 1.100 gy, two samples of surviving bacteria were again irradiated by doses up to 2.5000 Gy. The bacteria not previously irradiated were throughly killed by means of doses of 1.100 Gy. Salmonella enteritidis var. typhimurium was inactivated by means of 1.200 and 1.900 Gy doses. It was concluded that 60-Cobalt gamma radiation minimal lethal dose to Salmonella enteritidis var. typhimurium is 1.200 Gy; the re-irradiation to the survivors prompts the forthcoming of more resistant germs. (author) [pt

Elimination of Salmonella enterica serovar Typhimurium in artificially contaminated eggs through correct cooking and frying procedures

Directory of Open Access Journals (Sweden)

Geovana Dagostim Savi

2011-06-01

Full Text Available Salmonellosis is a serious foodborne disease associated with the presence of bacteria in eggs or foods containing raw eggs. However, the use of appropriate procedures of cooking and frying can eliminate this contamination. There are few studies on the elimination of contamination of Salmonella in hens' eggs through typical frying procedures, especially for Salmonella enterica serovar Typhimurium (or S. typhimurium. The aim of this study was to determine the appropriate conditions for cooking and frying hens' eggs artificially contaminated with S. typhimurium, making them free of bacterial contamination. Hens' eggs were artificially contaminated with S. typhimurium and subjected to various processes of cooking, frying and food preparation. It was observed that the minimum time necessary to eliminate contamination through cooking procedures is 5 minutes after the water starts boiling, and also that, cooking in the microwave oven complete eliminates the bacterial contamination. When the eggs were fried on both sides, keeping the yolk hard, a complete bacterial elimination was observed. Mayonnaise prepared with vinegar presented a decrease in bacterial colonies when compared mayonese prepared with lemon.

Prevalence and Characteristics of Salmonella Isolated from Free-Range Chickens in Shandong Province, China.

Science.gov (United States)

Zhao, Xiaonan; Gao, Yanxia; Ye, Chaoqun; Yang, Lingling; Wang, Tao; Chang, Weishan

2016-01-01

Compared with chickens raised in intensively managed breeding farms, free-range chickens in China are quite popular due to lower breeding density and less antibiotics usage. However, investigations about Salmonella enterica from free-range chickens are quite rare. The aim of the present study was to investigate prevalence and characteristics of Salmonella in free-range chickens in Shandong province, China. During the period of August and November 2015, 300 fresh fecal swabs from different broilers in three free-range chicken farms (100 samples per farm) were collected to isolate Salmonella , and then these isolates were subjected to serotyping, antibiotic sensitivity testing, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), and multilocus sequence typing (ST). A total of 38 Salmonella isolates (38/300, 12.7%) were recovered. The most common serotype was Enteritidis (81.6%), followed by Indiana (13.2%) and Typhimurium (5.3%). Twenty-two out of 38 isolates (57.9%) were resistant to ampicillin, the highest resistance rate, but resistance rates to cefazolin, cefotaxime, and ceftazidime were only 7.9%. The multidrug resistance (MDR) rate was 26.3%. Additionally, the Salmonella isolates could be classified into 25 genotypes by ERIC-PCR and were divided into three ST types (ST11, ST17, and ST19), with ST11 the highest isolation rate (81.6%). In summary, as with other poultry, free-ranging chickens may also serve as potential reservoir for antibiotic resistant Salmonella , thereby posing a threat to public health.

Specific Monoclonal Antibody Overcomes the Salmonella enterica Serovar Typhimurium's Adaptive Mechanisms of Intramacrophage Survival and Replication.

Directory of Open Access Journals (Sweden)

Swarmistha Devi Aribam

Full Text Available Salmonella-specific antibodies play an important role in host immunity; however, the mechanisms of Salmonella clearance by pathogen-specific antibodies remain to be completely elucidated since previous studies on antibody-mediated protection have yielded inconsistent results. These inconsistencies are at least partially attributable to the use of polyclonal antibodies against Salmonella antigens. Here, we developed a new monoclonal antibody (mAb-449 and identified its related immunogen that protected BALB/c mice from infection with Salmonella enterica serovar Typhimurium. In addition, these data indicate that the mAb-449 immunogen is likely a major protective antigen. Using in vitro infection studies, we also analyzed the mechanism by which mAb-449 conferred host protection. Notably, macrophages infected with mAb-449-treated S. Typhimurium showed enhanced pathogen uptake compared to counterparts infected with control IgG-treated bacteria. Moreover, these macrophages produced elevated levels of pro-inflammatory cytokine TNFα and nitric oxide, indicating that mAb-449 enhanced macrophage activation. Finally, the number of intracellular bacteria in mAb-449-activated macrophages decreased considerably, while the opposite was found in IgG-treated controls. Based on these findings, we suggest that, although S. Typhimurium has the potential to survive and replicate within macrophages, host production of a specific antibody can effectively mediate macrophage activation for clearance of intracellular bacteria.

Inhibition and inactivation of Salmonella typhimurium biofilms from polystyrene and stainless steel surfaces by essential oils and phenolic constituent carvacrol.

Science.gov (United States)

Soni, Kamlesh A; Oladunjoye, Ademola; Nannapaneni, Ramakrishna; Schilling, M Wes; Silva, Juan L; Mikel, Benjy; Bailey, R Hartford

2013-02-01

Persistence of Salmonella biofilms within food processing environments is an important source of Salmonella contamination in the food chain. In this study, essential oils of thyme and oregano and their antimicrobial phenolic constituent carvacrol were evaluated for their ability to inhibit biofilm formation and inactivate preformed Salmonella biofilms. A crystal violet staining assay and CFU measurements were utilized to quantify biofilm cell mass, with evaluating factors such as strain variation, essential oil type, their concentrations, exposure time, as well as biofilm formation surface. Of the three Salmonella strains, Salmonella Typhimurium ATCC 23564 and Salmonella Typhimurium ATCC 19585 produced stronger biofilms than Salmonella Typhimurium ATCC 14028. Biofilm formation by different Salmonella strains was 1.5- to 2-fold higher at 22°C than at 30 or 37°C. The presence of nonbiocidal concentrations of thyme oil, oregano oil, and phenolic carvacrol at 0.006 to 0.012% suppressed Salmonella spp. biofilm formation 2- to 4-fold, but could not completely eliminate biofilm formation. There was high correlation in terms of biofilm inactivation, as determined by the crystal violet-stained optical density (at a 562-nm wavelength) readings and the viable CFU counts. Reduction of biofilm cell mass was dependent on antimicrobial concentration. A minimum concentration of 0.05 to 0.1% of these antimicrobial agents was needed to reduce a 7-log CFU biofilm mass to a nondetectable level on both polystyrene and stainless steel surfaces within 1 h of exposure time.

A Descriptive Study of Human Salmonella Serotype Typhimurium Infections Reported in Ontario from 1990 to 1997

Directory of Open Access Journals (Sweden)

Michael W Ford

2003-01-01

Full Text Available BACKGROUND: Salmonella infections cause gastrointestinal and systemic diseases worldwide and are the leading causes of food-borne illnesses in North America (1-4. Salmonella serotype typhimurium (ST, in particular, is increasingly becoming a major public health concern because of its ability to acquire multiple resistant genes (5,6.

Salmonella enterica serovar Typhimurium ΔmsbB triggers exacerbated inflammation in Nod2 deficient mice.

Directory of Open Access Journals (Sweden)

Anne-Kathrin Claes

Full Text Available The intracellular pathogen Salmonella enterica serovar Typhimurium causes intestinal inflammation characterized by edema, neutrophil influx and increased pro-inflammatory cytokine expression. A major bacterial factor inducing pro-inflammatory host responses is lipopolysaccharide (LPS. S. Typhimurium ΔmsbB possesses a modified lipid A, has reduced virulence in mice, and is being considered as a potential anti-cancer vaccine strain. The lack of a late myristoyl transferase, encoded by MsbB leads to attenuated TLR4 stimulation. However, whether other host receptor pathways are also altered remains unclear. Nod1 and Nod2 are cytosolic pattern recognition receptors recognizing bacterial peptidoglycan. They play important roles in the host's immune response to enteric pathogens and in immune homeostasis. Here, we investigated how deletion of msbB affects Salmonella's interaction with Nod1 and Nod2. S. Typhimurium Δ msbB-induced inflammation was significantly exacerbated in Nod2-/- mice compared to C57Bl/6 mice. In addition, S. Typhimurium ΔmsbB maintained robust intestinal colonization in Nod2-/- mice from day 2 to day 7 p.i., whereas colonization levels significantly decreased in C57Bl/6 mice during this time. Similarly, infection of Nod1-/- and Nod1/Nod2 double-knockout mice revealed that both Nod1 and Nod2 play a protective role in S. Typhimurium ΔmsbB-induced colitis. To elucidate why S. Typhimurium ΔmsbB, but not wild-type S. Typhimurium, induced an exacerbated inflammatory response in Nod2-/- mice, we used HEK293 cells which were transiently transfected with pathogen recognition receptors. Stimulation of TLR2-transfected cells with S. Typhimurium ΔmsbB resulted in increased IL-8 production compared to wild-type S. Typhimurium. Our results indicate that S. Typhimurium ΔmsbB triggers exacerbated colitis in the absence of Nod1 and/or Nod2, which is likely due to increased TLR2 stimulation. How bacteria with "genetically detoxified" LPS

Effectiveness of radiation processing for elimination of Salmonella Typhimurium from minimally processed pineapple (Ananas comosus Merr.).

Science.gov (United States)

Shashidhar, Ravindranath; Dhokane, Varsha S; Hajare, Sachin N; Sharma, Arun; Bandekar, Jayant R

2007-04-01

The microbiological quality of market samples of minimally processed (MP) pineapple was examined. The effectiveness of radiation treatment in eliminating Salmonella Typhimurium from laboratory inoculated ready-to-eat pineapple slices was also studied. Microbiological quality of minimally processed pineapple samples from Mumbai market was poor; 8.8% of the samples were positive for Salmonella. D(10) (the radiation dose required to reduce bacterial population by 90%) value for S. Typhimurium inoculated in pineapple was 0.242 kGy. Inoculated pack studies in minimally processed pineapple showed that the treatment with a 2-kGy dose of gamma radiation could eliminate 5 log CFU/g of S. Typhimurium. The pathogen was not detected from radiation-processed samples up to 12 d during storage at 4 and 10 degrees C. The processing of market samples with 1 and 2 kGy was effective in improving the microbiological quality of these products.

An association of genotypes and antimicrobial resistance patterns among Salmonella isolates from pigs and humans in Taiwan.

Directory of Open Access Journals (Sweden)

Hung-Chih Kuo

Full Text Available We collected 110 Salmonella enterica isolates from sick pigs and determined their serotypes, genotypes using pulsed-field gel electrophoresis (PFGE, and antimicrobial susceptibility to 12 antimicrobials and compared the data with a collection of 18,280 isolates obtained from humans. The pig isolates fell into 12 common serovars for human salmonellosis in Taiwan; S. Typhimurium, S. Choleraesuis, S. Derby, S. Livingstone, and S. Schwarzengrund were the 5 most common serovars and accounted for a total of 84% of the collection. Of the 110 isolates, 106 (96% were multidrug resistant (MDR and 48 (44% had PFGE patterns found in human isolates. S. Typhimurium, S. Choleraesuis, and S. Schwarzengrund were among the most highly resistant serovars. The majority of the 3 serovars were resistant to 8-11 of the tested antimicrobials. The isolates from pigs and humans sharing a common PFGE pattern displayed identical or very similar resistance patterns and Salmonella strains that caused severe infection in pigs were also capable of causing infections in humans. The results indicate that pigs are one of the major reservoirs to human salmonellosis in Taiwan. Almost all of the pig isolates were MDR, which highlights the necessity of strictly regulating the use of antimicrobials in the agriculture sector in Taiwan.

An association of genotypes and antimicrobial resistance patterns among Salmonella isolates from pigs and humans in Taiwan.

Science.gov (United States)

Kuo, Hung-Chih; Lauderdale, Tsai-Ling; Lo, Dan-Yuan; Chen, Chiou-Lin; Chen, Pei-Chen; Liang, Shiu-Yun; Kuo, Jung-Che; Liao, Ying-Shu; Liao, Chun-Hsing; Tsao, Chi-Sen; Chiou, Chien-Shun

2014-01-01

We collected 110 Salmonella enterica isolates from sick pigs and determined their serotypes, genotypes using pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility to 12 antimicrobials and compared the data with a collection of 18,280 isolates obtained from humans. The pig isolates fell into 12 common serovars for human salmonellosis in Taiwan; S. Typhimurium, S. Choleraesuis, S. Derby, S. Livingstone, and S. Schwarzengrund were the 5 most common serovars and accounted for a total of 84% of the collection. Of the 110 isolates, 106 (96%) were multidrug resistant (MDR) and 48 (44%) had PFGE patterns found in human isolates. S. Typhimurium, S. Choleraesuis, and S. Schwarzengrund were among the most highly resistant serovars. The majority of the 3 serovars were resistant to 8-11 of the tested antimicrobials. The isolates from pigs and humans sharing a common PFGE pattern displayed identical or very similar resistance patterns and Salmonella strains that caused severe infection in pigs were also capable of causing infections in humans. The results indicate that pigs are one of the major reservoirs to human salmonellosis in Taiwan. Almost all of the pig isolates were MDR, which highlights the necessity of strictly regulating the use of antimicrobials in the agriculture sector in Taiwan.

Salmonella Typhimurium metabolism affects virulence in the host – A mini-review

DEFF Research Database (Denmark)

Herrero-fresno, Ana; Olsen, John Elmerdhahl

2018-01-01

Salmonella enterica remains an important food borne pathogen in all regions of the world with S. Typhimurium as one of the most frequent serovars causing food borne disease. Since the majority of human cases are caused by food of animal origin, there has been a high interest in understanding how S....... Typhimurium interacts with the animal host, mostly focusing on factors that allow it to breach host barriers and to manipulate host cells to the benefit of itself. Up to recently, such studies have ignored the metabolic factors that allow the bacteria to multiply in the host, but this is changing rapidly...

Higher Storage Temperature Causes Greater Salmonella enterica Serovar Typhimurium Internal Penetration of Artificially Contaminated, Commercially Available, Washed Free Range Eggs.

Science.gov (United States)

Whiley, Alice; Fallowfield, Howard; Ross, Kirstin; McEvoy, Vanessa; Whiley, Harriet

2016-07-01

Foodborne salmonellosis is a major public health concern, with contaminated eggs identified as a significant source of infection. In Australia, the most prevalent cause of salmonellosis from eggs is Salmonella enterica subsp. enterica serovar Typhimurium. This study explored the effect of temperature after 1, 7, 14, 21, and 28 days of storage on commercially available washed free range eggs, artificially contaminated with Salmonella Typhimurium on the external surface. At each time point, the external surface of the egg, the crushed eggshell, and the internal egg yolk and albumen were analyzed for Salmonella. After 28 days of storage, 25% of eggs stored at 4°C, 50% of eggs stored at 14°C, and 100% of eggs stored at 23 and 35°C were internally contaminated with Salmonella. After 1 day of storage, more than 50% of all eggs had Salmonella present in the crushed shell after the external surface had been disinfected with ethanol. This is the first study to demonstrate that refrigeration reduced the potential for Salmonella Typhimurium to penetrate the eggshell membrane and internally contaminate table eggs commercially available in Australia. It also suggests that the processes of cracking eggs may be a source of cross-contamination within the kitchen.

Immuno-capture and in situ detection of Salmonella typhimurium on a novel microfluidic chip

Energy Technology Data Exchange (ETDEWEB)

Wang, Renjie, E-mail: 1058464972@qq.com [College of Chemistry and Chemical Engineering, Chongqing University, No. 174, St. Shazheng, Shapingba District, Chongqing (China); Ni, Yanan, E-mail: 468885029@qq.com [College of Chemistry and Chemical Engineering, Chongqing University, No. 174, St. Shazheng, Shapingba District, Chongqing (China); Xu, Yi, E-mail: xuyibbd@sina.com [College of Chemistry and Chemical Engineering, Chongqing University, No. 174, St. Shazheng, Shapingba District, Chongqing (China); National Center for International Research of Micro/Nano-System and New Material Technology, No. 174, St. Shazhengjie, Shapingba District, Chongqing (China); Key Laboratory of Fundamental Science of Micro/Nano-Device and System Technology for National Defense, Chongqing (China); Jiang, Yan, E-mail: 919865356@qq.com [College of Chemistry and Chemical Engineering, Chongqing University, No. 174, St. Shazheng, Shapingba District, Chongqing (China); Dong, Chunyan, E-mail: 774176325@qq.com [College of Chemistry and Chemical Engineering, Chongqing University, No. 174, St. Shazheng, Shapingba District, Chongqing (China); Chuan, Na, E-mail: 814859441@qq.com [College of Chemistry and Chemical Engineering, Chongqing University, No. 174, St. Shazheng, Shapingba District, Chongqing (China)

2015-01-01

Highlights: • A novel microfluidic chip and a LIF microsystem were designed and fabricated. • Salmonella typhimurium was captured and labeled by specific immuno-capture on chip. • CdSe/ZnS quantum dots-labeled bacteria were detected by in situ analysis using LIF microsystem. • The proposed method has potential application in practice. - Abstract: The new method presented in this article achieved the goal of capturing Salmonella typhimurium via immunoreaction and rapid in situ detection of the CdSe/ZnS quantum dots (QDs) labeled S. typhimurium by self-assembly light-emitting diode-induced fluorescence detection (LIF) microsystem on a specially designed multichannel microfluidic chip. CdSe/ZnS QDs were used as fluorescent markers improving detection sensitivity. The microfluidic chip developed in this study was composed of 12 sample channels, 3 mixing zones, and 6 immune reaction zones, which also acted as fluorescence detection zones. QDs–IgG–primary antibody complexes were generated by mixing CdSe/ZnS QDs conjugated secondary antibody (QDs–IgG) and S. typhimurium antibody (primary antibody) in mixing zones. Then, the complexes went into immune reaction zones to label previously captured S. typhimurium in the sandwich mode. The capture rate of S. typhimurium in each detection zone was up to 70%. The enriched QDs-labeled S. typhimurium was detected using a self-assembly LIF microsystem. A good linear relationship was obtained in the range from 3.7 × 10 to 3.7 × 10{sup 5} cfu mL{sup −1} using the equation I = 0.1739 log (C) − 0.1889 with R{sup 2} = 0.9907, and the detection limit was down to 37 cfu mL{sup −1}. The proposed method of online immunolabeling with QDs for in situ fluorescence detection on the designed multichannel microfluidic chip had been successfully used to detect S. typhimurium in pork sample, and it has shown potential advantages in practice.

Salmonella enterica serovar Typhimurium lacking hfq gene confers protective immunity against murine typhoid.

Directory of Open Access Journals (Sweden)

Uday Shankar Allam

Full Text Available Salmonella enterica is an important enteric pathogen and its various serovars are involved in causing both systemic and intestinal diseases in humans and domestic animals. The emergence of multidrug-resistant strains of Salmonella leading to increased morbidity and mortality has further complicated its management. Live attenuated vaccines have been proven superior over killed or subunit vaccines due to their ability to induce protective immunity. Of the various strategies used for the generation of live attenuated vaccine strains, focus has gradually shifted towards manipulation of virulence regulator genes. Hfq is a RNA chaperon which mediates the binding of small RNAs to the mRNA and assists in post-transcriptional gene regulation in bacteria. In this study, we evaluated the efficacy of the Salmonella Typhimurium Δhfq strain as a candidate for live oral vaccine in murine model of typhoid fever. Salmonella hfq deletion mutant is highly attenuated in cell culture and animal model implying a significant role of Hfq in bacterial virulence. Oral immunization with the Salmonella hfq deletion mutant efficiently protects mice against subsequent oral challenge with virulent strain of Salmonella Typhimurium. Moreover, protection was induced upon both multiple as well as single dose of immunizations. The vaccine strain appears to be safe for use in pregnant mice and the protection is mediated by the increase in the number of CD4(+ T lymphocytes upon vaccination. The levels of serum IgG and secretory-IgA in intestinal washes specific to lipopolysaccharide and outer membrane protein were significantly increased upon vaccination. Furthermore, hfq deletion mutant showed enhanced antigen presentation by dendritic cells compared to the wild type strain. Taken together, the studies in murine immunization model suggest that the Salmonella hfq deletion mutant can be a novel live oral vaccine candidate.

Bovine salmonellosis in northeast of Iran: frequency, genetic fingerprinting and antimicrobial resistance patterns of Salmonella spp.

Science.gov (United States)

Halimi, Hessam A; Seifi, Hesam A; Rad, Mehrnaz

2014-01-01

To evaluate serovar and antimicrobial resistance patterns of Salmonella spp isolated from healthy, diseased and necropsied cows and calves in this observational study. Nineteen isolates recovered from feces and tissues of salmonellosis-affected animals of two commercial farms in north-east of Iran. In second part of the study, the two farms were sampled 4 times with an interval of 2 month. The samples included calves' feces, adult cows' feces, feeds, water, milk filters, and milk fed to calves. Five Salmonella were isolated from 332 fecal samples collected from calves and peri-parturient cows. No Salmonella was recovered from water, feed, milk filers and milk fed to calves. Salmonella Typhimurium was the most frequently isolate among all sero-groups. S. Dublin was only accounted for 8% (two out of 24) of isolates. Isolated Salmonella strains were used for the ERIC PCR DNA fingerprinting assay. Our results grouped Salmonella isolates into 3 clusters, suggesting that specific genotypes were responsible for each sero-group of Salmonella. The results also revealed diversity among Salmonella isolates in cluster III (sero-group B). Eighteen out of 19 Salmonella spp. were resistant to oxytetracycline. Five isolates out of 19 showed more than one drug resistance. Multi-drug resistance was seen only among Salmonella Typhimurium isolates. Enrofloxacin was the most susceptible antibiotic against all isolates in this study. The emergence of multiple antibiotic-resistant strains of Salmonella Typhimurium should be of great concern to the public. No correlation between ERIC fingerprinting and resistance patterns of Salmonella isolates was found, which indicates resistance to antimicrobial agents was not related to specific genetic background. Copyright © 2014 Asian Pacific Tropical Biomedical Magazine. Published by Elsevier B.V. All rights reserved.

59Fe uptake by Salmonella typhimurium strains of different epidemiological sources

International Nuclear Information System (INIS)

Rabsch, W.; Reissbrodt, R.

1985-01-01

All Salmonella typhimurium strains tested were able to use iron from transferrin. In buffered nutrient broth - poor in iron-content - the strains were tested in 59 FeCl 3 and 59 Fe-transferrin uptake in different growth phases. In the early log phase the strains are able to catch the 59 Fe 3+ in a very great amount as it is necessary for the growth. The content of 59 Fe per cell was in the late log phase reduced until to a value, which seen to be enough for growth. The acquisition of 59 Fe-transferrin between the early and late log phase tested by 4 S. typhimurium strains was different. (author)

Primary structure and mapping of the hupA gene of Salmonella typhimurium.

Science.gov (United States)

Higgins, N P; Hillyard, D

1988-01-01

In bacteria, the complex nucleoid structure is folded and maintained by negative superhelical tension and a set of type II DNA-binding proteins, also called histonelike proteins. The most abundant type II DNA-binding protein is HU. Southern blot analysis showed that Salmonella typhimurium contained two HU genes that corresponded to Escherichia coli genes hupA (encoding HU-2 protein) and hupB (encoding HU-1). Salmonella hupA was cloned, and the nucleotide sequence of the gene was determined. Comparison of hupA of E. coli and S. typhimurium revealed that the HU-2 proteins were identical and that there was high conservation of nucleotide sequences outside the coding frames of the genes. A 300-member genomic library of S. typhimurium was constructed by using random transposition of MudP, a specialized chimeric P22-Mu phage that packages chromosomal DNA unidirectionally from its insertion point. Oligonucleotide hybridization against the library identified one MudP insertion that lies within 28 kilobases of hupA; the MudP was 12% linked to purH at 90.5 min on the standard map. Plasmids expressing HU-2 had a surprising phenotype; they caused growth arrest when they were introduced into E. coli strains bearing a himA or hip mutation. These results suggest that IHF and HU have interactive roles in bacteria. Images PMID:3056912

Survival and Filamentation of Salmonella enterica Serovar Enteritidis PT4 and Salmonella enterica Serovar Typhimurium DT104 at Low Water Activity

Science.gov (United States)

Mattick, K. L.; Jørgensen, F.; Legan, J. D.; Cole, M. B.; Porter, J.; Lappin-Scott, H. M.; Humphrey, T. J.

2000-01-01

In this study we investigated the long-term survival of and morphological changes in Salmonella strains at low water activity (aw). Salmonella enterica serovar Enteritidis PT4 and Salmonella enterica serovar Typhimurium DT104 survived at low aw for long periods, but minimum humectant concentrations of 8% NaCl (aw, 0.95), 96% sucrose (aw, 0.94), and 32% glycerol (aw, 0.92) were bactericidal under most conditions. Salmonella rpoS mutants were usually more sensitive to bactericidal levels of NaCl, sucrose, and glycerol. At a lethal aw, incubation at 37°C resulted in more rapid loss of viability than incubation at 21°C. At aw values of 0.93 to 0.98, strains of S. enterica serovar Enteritidis and S. enterica serovar Typhimurium formed filaments, some of which were at least 200 μm long. Filamentation was independent of rpoS expression. When the preparations were returned to high-aw conditions, the filaments formed septa, and division was complete within approximately 2 to 3 h. The variable survival of Salmonella strains at low aw highlights the importance of strain choice when researchers produce modelling data to simulate worst-case scenarios or conduct risk assessments based on laboratory data. The continued increase in Salmonella biomass at low aw (without a concomitant increase in microbial count) would not have been detected by traditional microbiological enumeration tests if the tests had been performed immediately after low-aw storage. If Salmonella strains form filaments in food products that have low aw values (0.92 to 0.98), there are significant implications for public health and for designing methods for microbiological monitoring. PMID:10742199

Combining predictive microbiology with cold-enrichment in minced pork for quantifying low levels of Salmonella Typhimurium DT104

DEFF Research Database (Denmark)

Møller, C. O. A.; Aabo, Søren; Hansen, Tina Beck

2013-01-01

underestimation of the Salmonella count after enrichment. Whether competition between these species also took place in the meat during enrichment is not known. However, as the observed levels of the competitive species were below 5.5 log10-units it is questionable whether interaction with S. Typhimurium DT104......OBJECTIVE For determination of Salmonella concentration in meat various methods can be used depending on the expected level. When higher levels (102 to 103 bacteria or more per g) are anticipated, plate count techniques using selective agars, i.e. XLD, are appropriate whereas for low numbers (3......, the objective of this study was to evaluate the possibility to carry out a Salmonella enrichment step in the meat itself and use the two species interaction model, presented by Møller et al. (2013), for quantifying levels of Salmonella Typhimurium DT104 in minced pork. METHODS A total number of 101 minced pork...

Role of nitric oxide in Salmonella typhimurium-mediated cancer cell killing

International Nuclear Information System (INIS)

Barak, Yoram; Schreiber, Frank; Thorne, Steve H; Contag, Christopher H; DeBeer, Dirk; Matin, A

2010-01-01

Bacterial targeting of tumours is an important anti-cancer strategy. We previously showed that strain SL7838 of Salmonella typhimurium targets and kills cancer cells. Whether NO generation by the bacteria has a role in SL7838 lethality to cancer cells is explored. This bacterium has the mechanism for generating NO, but also for decomposing it. Mechanism underlying Salmonella typhimurium tumour therapy was investigated through in vitro and in vivo studies. NO measurements were conducted either by chemical assays (in vitro) or using Biosensors (in vivo). Cancer cells cytotoxic assay were done by using MTS. Bacterial cell survival and tumour burden were determined using molecular imaging techniques. SL7838 generated nitric oxide (NO) in anaerobic cell suspensions, inside infected cancer cells in vitro and in implanted 4T1 tumours in live mice, the last, as measured using microsensors. Thus, under these conditions, the NO generating pathway is more active than the decomposition pathway. The latter was eliminated, in strain SL7842, by the deletion of hmp- and norV genes, making SL7842 more proficient at generating NO than SL7838. SL7842 killed cancer cells more effectively than SL7838 in vitro, and this was dependent on nitrate availability. This strain was also ca. 100% more effective in treating implanted 4T1 mouse tumours than SL7838. NO generation capability is important in the killing of cancer cells by Salmonella strains

Comparison of SNP-based subtyping workflows for bacterial isolates using WGS data, applied to Salmonella enterica serotype Typhimurium and serotype 1,4,[5],12:i:-

Science.gov (United States)

Saltykova, Assia; Wuyts, Véronique; Mattheus, Wesley; Bertrand, Sophie; Roosens, Nancy H. C.; Marchal, Kathleen

2018-01-01

Whole genome sequencing represents a promising new technology for subtyping of bacterial pathogens. Besides the technological advances which have pushed the approach forward, the last years have been marked by considerable evolution of the whole genome sequencing data analysis methods. Prior to application of the technology as a routine epidemiological typing tool, however, reliable and efficient data analysis strategies need to be identified among the wide variety of the emerged methodologies. In this work, we have compared three existing SNP-based subtyping workflows using a benchmark dataset of 32 Salmonella enterica subsp. enterica serovar Typhimurium and serovar 1,4,[5],12:i:- isolates including five isolates from a confirmed outbreak and three isolates obtained from the same patient at different time points. The analysis was carried out using the original (high-coverage) and a down-sampled (low-coverage) datasets and two different reference genomes. All three tested workflows, namely CSI Phylogeny-based workflow, CFSAN-based workflow and PHEnix-based workflow, were able to correctly group the confirmed outbreak isolates and isolates from the same patient with all combinations of reference genomes and datasets. However, the workflows differed strongly with respect to the SNP distances between isolates and sensitivity towards sequencing coverage, which could be linked to the specific data analysis strategies used therein. To demonstrate the effect of particular data analysis steps, several modifications of the existing workflows were also tested. This allowed us to propose data analysis schemes most suitable for routine SNP-based subtyping applied to S. Typhimurium and S. 1,4,[5],12:i:-. Results presented in this study illustrate the importance of using correct data analysis strategies and to define benchmark and fine-tune parameters applied within routine data analysis pipelines to obtain optimal results. PMID:29408896

A Salmonella Typhimurium-Typhi genomic chimera: a model to study Vi polysaccharide capsule function in vivo.

Directory of Open Access Journals (Sweden)

Angela M Jansen

2011-07-01

Full Text Available The Vi capsular polysaccharide is a virulence-associated factor expressed by Salmonella enterica serotype Typhi but absent from virtually all other Salmonella serotypes. In order to study this determinant in vivo, we characterised a Vi-positive S. Typhimurium (C5.507 Vi(+, harbouring the Salmonella pathogenicity island (SPI-7, which encodes the Vi locus. S. Typhimurium C5.507 Vi(+ colonised and persisted in mice at similar levels compared to the parent strain, S. Typhimurium C5. However, the innate immune response to infection with C5.507 Vi(+ and SGB1, an isogenic derivative not expressing Vi, differed markedly. Infection with C5.507 Vi(+ resulted in a significant reduction in cellular trafficking of innate immune cells, including PMN and NK cells, compared to SGB1 Vi(- infected animals. C5.507 Vi(+ infection stimulated reduced numbers of TNF-α, MIP-2 and perforin producing cells compared to SGB1 Vi(-. The modulating effect associated with Vi was not observed in MyD88(-/- and was reduced in TLR4(-/- mice. The presence of the Vi capsule also correlated with induction of the anti-inflammatory cytokine IL-10 in vivo, a factor that impacted on chemotaxis and the activation of immune cells in vitro.

Resveratrol induces membrane and DNA disruption via pro-oxidant activity against Salmonella typhimurium.

Science.gov (United States)

Lee, Wonjong; Lee, Dong Gun

2017-07-22

Resveratrol is a flavonoid found in various plants including grapes, which has been reported to be active against various pathogenic bacteria. However, antibacterial effects and mechanisms via pro-oxidant property of resveratrol remain unknown and speculative. This research investigated antibacterial mechanism of resveratrol against a food-borne human pathogen Salmonella typhimurium, and confirmed the cell death associated oxidative damage. Resveratrol increased outer membrane permeability and membrane depolarization. It also was observed DNA injury responses such as DNA fragmentation, increasing DNA contents and cell division inhibition. Intracellular ROS accumulation, GSH depletion and significant increased malondialdehyde levels were confirmed, which indicated pro-oxidant activity of resveratrol and oxidative stress. Furthermore, the observed lethal damages were reduced by antioxidant N-acetylcysteine treatment supported the view that resveratrol-induced oxidative stress stimulated S. typhimurium cell death. In conclusion, this study expands understanding on role of pro-oxidant property and insight into previously unrecognized oxygen-dependent anti-Salmonella mechanism on resveratrol. Copyright © 2017 Elsevier Inc. All rights reserved.

Lactobacillus bulgaricus, Lactobacillus rhamnosus and Lactobacillus paracasei Attenuate Salmonella Enteritidis, Salmonella Heidelberg and Salmonella Typhimurium Colonization and Virulence Gene Expression In Vitro.

Science.gov (United States)

Muyyarikkandy, Muhammed Shafeekh; Amalaradjou, Mary Anne

2017-11-09

Salmonella Enteritidis (SE), Salmonella Typhimurium (ST), and Salmonella Heidelberg (SH) have been responsible for numerous outbreaks associated with the consumption of poultry meat and eggs. Salmonella colonization in chicken is characterized by initial attachment to the cecal epithelial cells (CEC) followed by dissemination to the liver, spleen, and oviduct. Since cecal colonization is critical to Salmonella transmission along the food chain continuum, reducing this intestinal association could potentially decrease poultry meat and egg contamination. Hence, this study investigated the efficacy of Lactobacillus delbreuckii sub species bulgaricus (NRRL B548; LD), Lactobacillus paracasei (DUP-13076; LP), and Lactobacillus rhamnosus (NRRL B442; LR) in reducing SE, ST, and SH colonization in CEC and survival in chicken macrophages. Additionally, their effect on expression of Salmonella virulence genes essential for cecal colonization and survival in macrophages was evaluated. All three probiotics significantly reduced Salmonella adhesion and invasion in CEC and survival in chicken macrophages ( p < 0.05). Further, the probiotic treatment led to a significant reduction in Salmonella virulence gene expression ( p < 0.05). Results of the study indicate that LD, LP, and LR could potentially be used to control SE, ST, and SH colonization in chicken. However, these observations warrant further in vivo validation.

Lactobacillus bulgaricus, Lactobacillus rhamnosus and Lactobacillus paracasei Attenuate Salmonella Enteritidis, Salmonella Heidelberg and Salmonella Typhimurium Colonization and Virulence Gene Expression In Vitro

Directory of Open Access Journals (Sweden)

Muhammed Shafeekh Muyyarikkandy

2017-11-01

Full Text Available Salmonella Enteritidis (SE, Salmonella Typhimurium (ST, and Salmonella Heidelberg (SH have been responsible for numerous outbreaks associated with the consumption of poultry meat and eggs. Salmonella colonization in chicken is characterized by initial attachment to the cecal epithelial cells (CEC followed by dissemination to the liver, spleen, and oviduct. Since cecal colonization is critical to Salmonella transmission along the food chain continuum, reducing this intestinal association could potentially decrease poultry meat and egg contamination. Hence, this study investigated the efficacy of Lactobacillus delbreuckii sub species bulgaricus (NRRL B548; LD, Lactobacillus paracasei (DUP-13076; LP, and Lactobacillus rhamnosus (NRRL B442; LR in reducing SE, ST, and SH colonization in CEC and survival in chicken macrophages. Additionally, their effect on expression of Salmonella virulence genes essential for cecal colonization and survival in macrophages was evaluated. All three probiotics significantly reduced Salmonella adhesion and invasion in CEC and survival in chicken macrophages (p < 0.05. Further, the probiotic treatment led to a significant reduction in Salmonella virulence gene expression (p < 0.05. Results of the study indicate that LD, LP, and LR could potentially be used to control SE, ST, and SH colonization in chicken. However, these observations warrant further in vivo validation.

Influence of Temperature and Predation on Survival of Salmonella enterica Serovar Typhimurium and Expression of invA in Soil and Manure-Amended Soil▿

Science.gov (United States)

García, R.; Bælum, J.; Fredslund, L.; Santorum, P.; Jacobsen, C. S.

2010-01-01

The effects of three temperatures (5, 15, and 25°C) on the survival of Salmonella enterica serovar Typhimurium in topsoil were investigated in small microcosms by three different techniques: plate counting, invA gene quantification, and invA mRNA quantification. Differences in survival were related to the effect of protozoan predation. Tetracycline-resistant Salmonella serovar Typhimurium was inoculated into soil and manure-amended soil at 1.5 × 108 cells g soil−1. Population densities were determined by plate counting and by molecular methods and monitored for 42 days. Simultaneous extraction of RNA and DNA, followed by quantitative PCR, was used to investigate invA gene levels and expression. Analysis by these three techniques showed that Salmonella serovar Typhimurium survived better at 5°C. Comparing DNA and CFU levels, significantly higher values were determined by DNA-based techniques. invA mRNA levels showed a fast decrease in activity, with no detectable mRNA after an incubation period of less than 4 days in any of the soil scenarios. A negative correlation was found between Salmonella serovar Typhimurium CFU levels and protozoan most probable numbers, and we propose the role of the predator-prey interaction as a factor to explain the die-off of the introduced strain by both culture- and DNA quantification-based methods. The results indicate that temperature, manure, and protozoan predation are important factors influencing the survival of Salmonella serovar Typhimurium in soil. PMID:20562283

Quantification of contamination of lettuce by GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium

NARCIS (Netherlands)

Franz, Eelco; Visser, Anna A; Van Diepeningen, Anne D; Klerks, Michel M; Termorshuizen, Aad J; van Bruggen, Ariena H C

The primary objective of this study was to determine the possibility of internalization of GFP-expressing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium (S. Typhimurium) strains MAE 110 (multi-cellular morphology) and 119 (wild type morphology) into lettuce seedlings (Lactuca

Genetic Relatedness of Salmonella Serovars Isolated from Catfish (Clarias gariepinus) and Tilapia (Tilapia mossambica) Obtained from Wet Markets and Ponds in Penang, Malaysia.

Science.gov (United States)

Budiati, Titik; Rusul, Gulam; Wan-Abdullah, Wan Nadiah; Chuah, Li-Oon; Ahmad, Rosma; Thong, Kwai Lin

2016-04-01

A total of 43 Salmonella enterica isolates belonging to different serovars (Salmonella Albany, Salmonella Agona, Salmonella Corvallis, Salmonella Stanley, Salmonella Typhimurium, Salmonella Mikawasima, and Salmonella Bovismorbificans) were isolated from catfish (Clarias gariepinus) and tilapia (Tilapia mossambica) obtained from nine wet markets and eight ponds in Penang, Malaysia. Thirteen, 19, and 11 isolates were isolated from 9 of 32 catfish, 14 of 32 tilapia, and 11 of 44 water samples, respectively. Fish reared in ponds were fed chicken offal, spoiled eggs, and commercial fish feed. The genetic relatedness of these Salmonella isolates was determined by random amplified polymorphic DNA PCR (RAPD-PCR) using primer OPC2, repetitive extragenic palindromic PCR (REP-PCR), and pulsed-field gel electrophoresis (PFGE). Composite analysis of the RAPD-PCR, REP-PCR, and PFGE results showed that the Salmonella serovars could be differentiated into six clusters and 15 singletons. RAPD-PCR differentiated the Salmonella isolates into 11 clusters and 10 singletons, while REP-PCR differentiated them into 4 clusters and 1 singleton. PFGE differentiated the Salmonella isolates into seven clusters and seven singletons. The close genetic relationship of Salmonella isolates from catfish or tilapia obtained from different ponds, irrespective of the type of feed given, may be caused by several factors, such as the quality of the water, density of fish, and size of ponds.

Structure of Salmonella typhimurium OMP Synthase in a Complete Substrate Complex

DEFF Research Database (Denmark)

Grubmeyer, Charles; Hansen, Michael Riis; Fedorov, Alexander A.

2012-01-01

Dimeric Salmonella typhimurium orotate phosphoribosyltransferase (OMP synthase, EC 2.4.2.10), a key enzyme in de novo pyrimidine nucleotide synthesis, has been cocrystallized in a complete substrate E·MgPRPP·orotate complex and the structure determined to 2.2 Å resolution. This structure resem...

Impact of phytopathogen infection and extreme weather stress on internalization of Salmonella Typhimurium in lettuce.

Science.gov (United States)

Ge, Chongtao; Lee, Cheonghoon; Nangle, Ed; Li, Jianrong; Gardner, David; Kleinhenz, Matthew; Lee, Jiyoung

2014-01-03

Internalization of human pathogens, common in many types of fresh produce, is a threat to human health since the internalized pathogens cannot be fully inactivated/removed by washing with water or sanitizers. Given that pathogen internalization can be affected by many environmental factors, this study was conducted to investigate the influence of two types of plant stress on the internalization of Salmonella Typhimurium in iceberg lettuce during pre-harvest. The stresses were: abiotic (water stress induced by extreme weather events) and biotic (phytopathogen infection by lettuce mosaic virus [LMV]). Lettuce with and without LMV infection were purposefully contaminated with green fluorescence protein-labeled S. Typhimurium on the leaf surfaces. Lettuce was also subjected to water stress conditions (drought and storm) which were simulated by irrigating with different amounts of water. The internalized S. Typhimurium in the different parts of the lettuce were quantified by plate count and real-time quantitative PCR and confirmed with a laser scanning confocal microscope. Salmonella internalization occurred under the conditions outlined above; however internalization levels were not significantly affected by water stress alone. In contrast, the extent of culturable S. Typhimurium internalized in the leafy part of the lettuce decreased when infected with LMV under water stress conditions and contaminated with high levels of S. Typhimurium. On the other hand, LMV-infected lettuce showed a significant increase in the levels of culturable bacteria in the roots. In conclusion, internalization was observed under all experimental conditions when the lettuce surface was contaminated with S. Typhimurium. However, the extent of internalization was only affected by water stress when lettuce was infected with LMV. © 2013.

Antimicrobial resistance, class 1 integrons, and genomic island 1 in Salmonella isolates from Vietnam.

Directory of Open Access Journals (Sweden)

An T T Vo

Full Text Available BACKGROUND: The objective was to investigate the phenotypic and genotypic resistance and the horizontal transfer of resistance determinants from Salmonella isolates from humans and animals in Vietnam. METHODOLOGY/PRINCIPAL FINDINGS: The susceptibility of 297 epidemiologically unrelated non-typhoid Salmonella isolates was investigated by disk diffusion assay. The isolates were screened for the presence of class 1 integrons and Salmonella genomic island 1 by PCR. The potential for the transfer of resistance determinants was investigated by conjugation experiments. Resistance to gentamicin, kanamycin, chloramphenicol, streptomycin, trimethoprim, ampicillin, nalidixic acid, sulphonamides, and tetracycline was found in 13 to 50% of the isolates. Nine distinct integron types were detected in 28% of the isolates belonging to 11 Salmonella serovars including S. Tallahassee. Gene cassettes identified were aadA1, aadA2, aadA5, bla(PSE-1, bla(OXA-30, dfrA1, dfrA12, dfrA17, and sat, as well as open reading frames with unknown functions. Most integrons were located on conjugative plasmids, which can transfer their antimicrobial resistance determinants to Escherichia coli or Salmonella Enteritidis, or with Salmonella Genomic Island 1 or its variants. The resistance gene cluster in serovar Emek identified by PCR mapping and nucleotide sequencing contained SGI1-J3 which is integrated in SGI1 at another position than the majority of SGI1. This is the second report on the insertion of SGI1 at this position. High-level resistance to fluoroquinolones was found in 3 multiresistant S. Typhimurium isolates and was associated with mutations in the gyrA gene leading to the amino acid changes Ser83Phe and Asp87Asn. CONCLUSIONS: Resistance was common among Vietnamese Salmonella isolates from different sources. Legislation to enforce a more prudent use of antibiotics in both human and veterinary medicine should be implemented by the authorities in Vietnam.

Child morbidity of salmonellosis and the level of resistance of clinical isolates of salmonella to antibacterial preparations in saint Petersburg

Directory of Open Access Journals (Sweden)

N. V. Gonchar

2015-01-01

Full Text Available The aim of the study was to study the dynamics of the incidence of salmonellosis children in St. Petersburg and phenotypic resistance of clinical isolates of S. Enteritidis and S. Typhimurium to antibiotics in recent years. Materials and methods. The incidence of salmonellosis children studied according to the report for the first nine months of Rospotrebnadzor in 2013–2014. Incidence of salmonellosis in the structure of bacterial intestinal infections caused by pathogens in children hospitalized in the Department of intestinal infections in 2013–2014, studied according to annual reports. Antibiotic sensitivity was studied 86 Salmonella isolates (S. Enteritidis strain 64 and strain 22 S. Typhimurium, isolated from patients in children 2010–2014. Used the method of serial microdilution broth. Salmonella isolates were divided into sensitive, resistant, intermediate sensitivity to antibiotics. The Results. Analysis of the incidence of salmonellosis children of St. Petersburg has revealed its decline in 2014 (109.2 compared to 2013 (123,9 but relatively long-term average level was an increase in incidence (107,6. In the structure of salmonellosis in children prevailed salmonellosis Group D. In hospitalized children in the structure of bacterial intestinal infections detected Excess of share of salmonellosis in 2014 (36,9±3,4% compared to 2013 (24,5±2,4%; p <0,01. A reduction in the frequency sensitivity of S. Enteritidis to ampicillin, cefepime, ceftazidime and chloramphenicol. Compared to S. Enteritidis S. Typhimurium isolates were more resistant to ceftazidime and ampicillin, but more sensitive to ciprofloxacin. Conclusion. Morbidity of salmonellosis in recent years characterized by a relatively long-term average increase of the level. In the structure of salmonellosis in children prevailed salmonellosis Group D. There was a reduction of sensitivity S. Enteritidis isolates to cephalosporins new generations, and S. Typhimurium isolates

Occurrence of extended-spectrum and AmpC β-lactamases in multiple drug resistant Salmonella isolates from clinical samples in Lagos, Nigeria

Directory of Open Access Journals (Sweden)

Akinyemi KO

2017-01-01

Full Text Available KO Akinyemi,1 Bamidele Abiodun Iwalokun,2 Akeeb O Bola Oyefolu,1 CO Fakorede1 1Department of Microbiology, Lagos State University, Ojo, 2Molecular Biology and Biotechnology Division, Nigerian Institute of Medical Research, Yaba, Lagos, Nigeria Purpose: Salmonella spp. are important foodborne pathogens exhibiting increasing resistance to antimicrobial drugs. Resistance to broad-spectrum β-lactams, mediated by extended-spectrum β-lactamase (ESBL and AmpC β-lactamase enzymes is fast spreading and has had negative impacts on the clinical outcomes, particularly on third-generation cephalosporins. This study investigated the carriage of AmpC gene among multidrug-resistant Salmonella spp. from Lagos, Nigeria. Methods: Forty Salmonella spp. from clinical samples (S. typhi = 13; S. typhimurium = 10; S. enteritidis = 8; S. choleraesuis = 5; S. paratyphi = 4 were subjected to in vitro susceptibility test by disk diffusion methods. Isolates that were resistant to cefoxitin and third-generation cephalosporins were screened for ESBL (Double Disk Synergy Test Method and AmpC enzyme (AmpC disk test production. Detection of AmpC fox gene was carried out by polymerase chain reaction. Results: Thirty-two (80% of the Salmonella isolates were cefoxitin resistant. Plasmid-mediated AmpC β-lactamase and ESBL enzymes were recorded in 10/40 (25% and 16/40 (40% of the Salmonella isolates, respectively. Specifically, 16/40 (40% of the Salmonella isolates possessed 380 bp AmpC fox gene, with the highest occurrence found in S. typhi strains (43.8% followed by S. typhimurium (25%. There was no AmpC fox gene detected in S. paratyphi strains. Interestingly, coproduction of enzymes occurred in some of the isolates, raising fears of resistance to a multitude of antibiotics in the treatment of bacterial infections. Conclusion: Emergence of AmpC β-lactamase–producing Salmonella isolates in our environment was recorded for the first time, raising concern on increased

Genomic analysis of $\\textit{Salmonella enterica}$ serovar Typhimurium from wild passerines in England and Wales

OpenAIRE

Mather, Alison E; Lawson, Becki; de, Pinna Elizabeth; Wigley, Paul; Parkhill, Julian; Thomson, Nicholas R; Page, Andrew J; Holmes, Mark Adrian; Paterson, Gavin K

2016-01-01

Passerine salmonellosis is a well-recognised disease of birds in the order Passeriformes, including common songbirds such as finches and sparrows, caused by infection with $\\textit{Salmonella enterica}$ serovar Typhimurium. Previous research has suggested that some subtypes of S. Typhimurium – definitive phage types (DT) 40, 56 variant, and 160 – are host-adapted to passerines, and that these birds may represent a reservoir of infection for humans and other animals. Here, we have used whole g...

Mutagenicity of irradiated solutions of nuclei acid bases and nucleosides in Salmonella typhimurium

International Nuclear Information System (INIS)

Wilmer, J.; Schubert, J.

1981-01-01

Solutions of nucleic acid bases, nucleosides and a nucleotide, saturated with either N 2 , N 2 O or O 2 , were irradiated and tested for mutagenicity towards Salmonella typhimurium, with and without pre-incubation. Irradiated solutions of the nuclei acid bases were all non-mutagenic. Irradiated solutions of the nucleosides showed mutagenicity in S. typhimurium TA100 (pre-incubation assay). Generally, the mutagenicity followed the order: N 2 O > N 2 > O 2 . The results show that the formation of mutagenic radiolytic products is initiated by attack of mainly solutions of the nucleotide thymidine-5'-monophosphate, no mutagenicity could be detected. (orig.)

Inactivation of Enterobacter sakazakii, Bacillus cereus, and Salmonella typhimurium in powdered weaning food by electron-beam irradiation

Science.gov (United States)

Hong, Yun-Hee; Park, Ji-Yong; Park, Jong-Hyun; Chung, Myong-Soo; Kwon, Ki-Sung; Chung, Kyungsook; Won, Misun; Song, Kyung-Bin

2008-09-01

Inactivation of Enterobacter sakazakii, Bacillus cereus, and Salmonella typhimurium were evaluated in powdered weaning food using electron-beam irradiation. E. sakazakii, B. cereus, and S. typhimurium were eliminated by irradiation at 16, 8, and 8 kGy, respectively. The D10-vlaues of E. sakazakii, B. cereus, and S. typhimurium inoculated on powdered weaning food were 4.83, 1.22, and 0.98 kGy, respectively. The results suggest that electron-beam irradiation should inhibit the growth of pathogenic bacteria on baby food without impairing qualities.

Inactivation of Enterobacter sakazakii, Bacillus cereus, and Salmonella typhimurium in powdered weaning food by electron-beam irradiation

Energy Technology Data Exchange (ETDEWEB)

Hong, Yun-Hee [Department of Food Science and Technology, College of Agriculture and Life Science, Chungnam National University, Yuseong-Gu, Daejeon 305-764 (Korea, Republic of); Park, Ji-Yong [Department of Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of); Park, Jong-Hyun [Department of Food Science and Biotechnology, Kyungwon University, Sungnam 461-701 (Korea, Republic of); Chung, Myong-Soo [Department of Food Science, Ehwa Women' s University, Seoul 120-750 (Korea, Republic of); Kwon, Ki-Sung [Center for Food safety Evaluation, Korea Food and Drug Administration, Seoul 122-704 (Korea, Republic of); Chung, Kyungsook; Won, Misun [Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-333 (Korea, Republic of); Song, Kyung-Bin [Department of Food Science and Technology, College of Agriculture and Life Science, Chungnam National University, Yuseong-Gu, Daejeon 305-764 (Korea, Republic of)], E-mail: kbsong@cnu.ac.kr

2008-09-15

Inactivation of Enterobacter sakazakii, Bacillus cereus, and Salmonella typhimurium were evaluated in powdered weaning food using electron-beam irradiation. E. sakazakii, B. cereus, and S. typhimurium were eliminated by irradiation at 16, 8, and 8 kGy, respectively. The D{sub 10}-vlaues of E. sakazakii, B. cereus, and S. typhimurium inoculated on powdered weaning food were 4.83, 1.22, and 0.98 kGy, respectively. The results suggest that electron-beam irradiation should inhibit the growth of pathogenic bacteria on baby food without impairing qualities.

Inactivation of Enterobacter sakazakii, Bacillus cereus, and Salmonella typhimurium in powdered weaning food by electron-beam irradiation

International Nuclear Information System (INIS)

Hong, Yun-Hee; Park, Ji-Yong; Park, Jong-Hyun; Chung, Myong-Soo; Kwon, Ki-Sung; Chung, Kyungsook; Won, Misun; Song, Kyung-Bin

2008-01-01

Inactivation of Enterobacter sakazakii, Bacillus cereus, and Salmonella typhimurium were evaluated in powdered weaning food using electron-beam irradiation. E. sakazakii, B. cereus, and S. typhimurium were eliminated by irradiation at 16, 8, and 8 kGy, respectively. The D 10 -vlaues of E. sakazakii, B. cereus, and S. typhimurium inoculated on powdered weaning food were 4.83, 1.22, and 0.98 kGy, respectively. The results suggest that electron-beam irradiation should inhibit the growth of pathogenic bacteria on baby food without impairing qualities

Investigation of the role of genes encoding zinc exporters zntA, zitB, and fieF during Salmonella typhimurium infection

DEFF Research Database (Denmark)

Huang, Kaisong; Wang, Dan; Frederiksen, Rikki F.

2018-01-01

The transition metal zinc is involved in crucial biological processes in all living organisms and is essential for survival of Salmonella in the host. However, little is known about the role of genes encoding zinc efflux transporters during Salmonella infection. In this study, we constructed...... deletion mutants for genes encoding zinc exporters (zntA, zitB, and fieF) in the wild-type (WT) strain Salmonella enterica serovar Typhimurium (S. Typhimurium) 4/74. The mutants 4/74ΔzntA and 4/74ΔzntA/zitB exhibited a dramatic growth delay and abrogated growth ability, respectively, in Luria Bertani...... medium supplemented with 0.25 mM ZnCl2 or 1.5 mM CuSO4 compared to the WT strain. In order to investigate the role of genes encoding zinc exporters on survival of S. Typhimurium inside cells, amoeba and macrophage infection models were used. No significant differences in uptake or survival were detected...

Resuscitation of the viable but non-culturable state of Salmonella enterica serovar Oranienburg by recombinant resuscitation-promoting factor derived from Salmonella Typhimurium strain LT2.

Science.gov (United States)

Panutdaporn, N; Kawamoto, K; Asakura, H; Makino, S-I

2006-02-15

A gene encoding the resuscitation-promoting factor (Rpf) from Salmonella Typhimurium LT2 was cloned and characterized. The amino acid sequence encoded by S. Typhimurium LT2 rpf gene shares 24.2% homology with Micrococcus luteus Rpf, which is secreted by growing cells, and required to resuscitate from viable but non-culturable (VNC) state. The S. Typhimurium LT2 rpf gene is 696 bp long, and shared a conserved segment with Salmonella enterica serovar Oranienburg (99.4%). Recombinant Rpf (rRpf) proteins of S. Typhimurium LT2 after expression in E. coli BL21 harboring the pET15-b plasmid was approximately 25 kDa. Since S. Oranienburg cells are relatively quick to enter the VNC state just after incubating in the presence of 7% NaCl at 37 degrees C for 3 days, we evaluated the biological effect of rRpf by using S. Oranienburg VNC cells. The rRpf not only promoted proliferation but also induced resuscitation of VNC cells to the culturable state in a dose-dependent manner. Therefore, rRpf may be useful for detection of bacterial contaminants present in the VNC form in food samples and the environment.

Case Report: Salmonella lung infection | Ohanu | International ...

African Journals Online (AJOL)

A case of an 84 year old man admitted because of fever, abdominal discomfort, weakness, past history of cough wheezing and abuse of prednisolone and Erythromycin. He had Bronchopneumonia and diabetes. Salmonella typhimurium was isolated from both his sputum and blood while stool was negative for salmonella.

Lack of specific hybridization between the lep genes of Salmonella typhimurium and Bacillus licheniformis

NARCIS (Netherlands)

van Dijl, J M; Jong, de Anne; Smith, H; Bron, Sierd; Venema, G

1991-01-01

This paper describes an attempt to clone the Bacillus licheniformis lep gene, encoding signal peptidase, using the Salmonella typhimurium lep gene as a hybridization probe. Although a hybridizing fragment was obtained, DNA sequence analysis indicated that it did not contain the lep gene. Instead,

Survival of Salmonella Typhimurium in poultry-based meat preparations during grilling, frying and baking.

Science.gov (United States)

Roccato, Anna; Uyttendaele, Mieke; Cibin, Veronica; Barrucci, Federica; Cappa, Veronica; Zavagnin, Paola; Longo, Alessandra; Ricci, Antonia

2015-03-16

The burden of food-borne diseases still represents a threat to public health; in 2012, the domestic setting accounted for 57.6% of strong-evidence EU food-borne Salmonella outbreaks. Next to cross-contamination, inadequate cooking procedure is considered as one of the most important factors contributing to food-borne illness. The few studies which have assessed the effect of domestic cooking on the presence and numbers of pathogens in different types of meat have shown that consumer-style cooking methods can allow bacteria to survive and that the probability of eating home-cooked poultry meat that still contains surviving bacteria after heating is higher than previously assumed. Thus, the main purpose of this study was to reproduce and assess the effect of several types of cooking treatments (according to label instructions and not following label instructions) on the presence and numbers of Salmonella Typhimurium DT 104 artificially inoculated in five types of poultry-based meat preparations (burgers, sausages, ready-to-cook-kebabs, quail roulades and extruded roulades) that are likely to be contaminated by Salmonella. Three contamination levels (10 cfu/g; 100 cfu/g and 1000 cfu/g) and three cooking techniques (grilling, frying and baking) were applied. Cooking treatments performed according to label instructions eliminated Salmonella Typhimurium (absence per 25g) for contamination levels of 10 and 100 cfu/g but not for contamination levels of 1000 cfu/g. After improper cooking, 26 out of 78 samples were Salmonella-positive, and 23 out of these 26 samples were artificially contaminated with bacterial loads between 100 and 1000 cfu/g. Nine out of 26 samples provided quantifiable results with a minimum level of 1.4MPN/g in kebabs (initial inoculum level: 100 cfu/g) after grilling and a maximum level of 170MPN/g recorded in sausages (initial inoculum level: 1000 cfu/g) after grilling. Kebabs were the most common Salmonella-positive meat product after cooking

Effect of the oyster contaminated in Salmonella typhimurium and Escherichia coli; Efecto del ostion contaminado en Salmonella typhimurium y Escherichia coli

Energy Technology Data Exchange (ETDEWEB)

Brena V, M

1992-08-15

In this work the effect of the oyster contaminated in the species of bacteria better studied by its genetic composition these are the Salmonella typhimurium and the Escherichia coli and that its have been starting point for later radiobiological studies in superior organisms. Of this its have arisen two general lines. The research about the genotoxic effect of substances or mixtures in bacteria with the collaboration of the groups of Drosophila and X-ray Fluorescence analysis and on the other hand the study of the low doses of radiation also in bacteria. It is also treated the topic about the genetic effect of aromatic hydrocarbons in different biological systems. (Author)

Influence of Natural Organic Matter on Attachment Kinetics of Salmonella Typhimurium

Science.gov (United States)

Chowdhury, I.; Zorlu, O.; Hill, J. E.; Walker, S. L.

2011-12-01

Salmonella enterica serovar Typhimurium is one of the most common and virulent bacterial pathogens, usually found in food and water. This waterborne pathogen has been attributed to causing gastroenteritis and typhoid fever, leading to 16 million cases and over half a million deaths worldwide each year. Natural organic matter (NOM) is ubiquitous in environment and previous work has shown NOM to enhance the stability and transport of bacteria cells; hence NOM will certainly interact with Salmonella and affect its transport in environment. The objective of this study was to investigate the influence of NOM (Suwannee River humic acid standard II, SRHA) on the attachment kinetics of a model Salmonella (Salmonella enterica serovar Typhimurium SA5983) to glass. The transport study was conducted in a parallel plate flow chamber using fluorescent microscope to visualize the bacterial cells, which were tagged with green fluorescent protein (GFP). The solution pH was unadjusted, and the flow rate through parallel plate channel was 0.1 mL/min to simulate groundwater conditions. Parameters varied in this study were NOM presence, ion valence (K+, Ca2+) as well as cell growth phase (mid-exponential and late-exponential growth phases). These parameters were chosen because ion valence may alter the NOM conformation and capacity for bridging, as well growth phase impacts the cellular surface chemistry. Extensive characterization of the bacterial cells was conducted including measurements of electrophoretic mobility, hydrophobicity, acidity, surface charge density and extracellular polymeric substance content. Additionally, electrokintic characterization was conducted for the glass. Preliminary results demonstrated the sensitivity of cell attachment to ionic valence and cell growth phase. Also the addition of NOM reduced the attachment of the Salmonella cells significantly under all of these conditions. Without NOM, attachment efficiencies (α) in KCl were similar at both growth

Detection of cell surface hydrophobicity, biofilm and fimbirae genes in salmonella isolated from tunisian clinical and poultry meat.

Science.gov (United States)

Ben Abdallah, Fethi; Lagha, Rihab; Said, Khaled; Kallel, Héla; Gharbi, Jawhar

2014-04-01

The aim of this study was to evaluate the ability of 15 serotypes of Salmonella to form biofilm on polystyrene, polyvinyl chloride (PVC) and glass surfaces. . Initially slime production was assessed on CRA agar and hydrophobicity of 20 Salmonella strains isolated from poultry and human and two Salmonella enterica serovar Typhimurium references strains was achieved by microbial adhesion to n-hexadecane. In addition, biofilm formation on polystyrene, PVC and glass surfaces was also investigated by using MTT and XTT colorimetric assay. Further, distribution of Salmonella enterotoxin (stn), Salmonella Enteritidis fimbrial (sef) and plasmid encoded fimbrial (pef) genes among tested strains was achieved by PCR. Salmonella strains developed red and white colonies on CRA and they are considered as hydrophilic with affinity values to n-hexadecane ranged between 0.29% and 29.55%. Quantitative biofilm assays showed that bacteria are able to form biofilm on polystyrene with different degrees and 54.54% of strains produce a strong biofilm on glass. In addition, all the strains form only a moderate (54.54%) and weak (40.91%) biofilm on PVC. PCR detection showed that only S. Enteritidis harbour Sef gene, whereas Pef and stn genes were detected in S. Kentucky, S. Amsterdam, S. Hadar, S. Enteritidis and S. Typhimurium. Salmonella serotypes are able to form biofilm on hydrophobic and hydrophilic industrial surfaces. Biofilm formation of Salmonella on these surfaces has an increased potential to compromise food safety and potentiate public health risk.

Determination method of inactivating minimal dose of gama radiation for Salmonella typhimurium

International Nuclear Information System (INIS)

Araujo, E.S.; Campos, H. de; Silva, D.M.

1979-01-01

A method for determination of minimal inactivating dose (MID) with Salmonella typhimurium is presented. This is a more efficient way to improve the irradiated vaccines. The MID found for S. thyphimurium 6.616 by binomial test was 0.55 MR. The method used allows to get a definite value for MID and requires less consumption of material, work and time in comparison with the usual procedure [pt

Growth potential of exponential- and stationary-phase Salmonella Typhimurium during sausage fermentation

DEFF Research Database (Denmark)

Birk, Tina; Henriksen, Sidsel; Müller, K.

2016-01-01

- and stationary-phase Salmonella Typhimurium (DT12 and DTU292) during freezing at − 18 °C and their subsequent growth potential during 72 h sausage fermentation at 25 °C. After 0, 7 and > 35 d of frozen storage, sausage batters were prepared with NaCl (3%) and NaNO2 (0, 100 ppm) and fermented with and without...

The Role of the st313-td Gene in Virulence of Salmonella Typhimurium ST313

DEFF Research Database (Denmark)

Herrero-Fresno, Ana; Wallrodt, Inke; Leekitcharoenphon, Pimlapas

2014-01-01

Multidrug-resistant Salmonella enterica serovar Typhimurium ST313 has emerged in sub-Saharan Africa causing severe infections in humans. Therefore, it has been speculated that this specific sequence type, ST313, carries factors associated with increased pathogenicity. We assessed the role in viru...

A long-lasting outbreak of Salmonella Typhimurium U323 associated with several pork products, Denmark, 2010

DEFF Research Database (Denmark)

Kuhn, K. G.; Sørensen, Gitte; Torpdahl, M.

2013-01-01

This paper shows that control of foodborne disease outbreaks may be challenging even after establishing the source of infection. An outbreak of Salmonella Typhimurium U323 infections occurred in Denmark from March to September 2010, involving 172 cases. Before the detection of human cases, severa...... ready-to-eat spreadable pork sausage (teewurst) as the source of a sub-outbreak (matched odds ratio 17, 95% confidence interval 2·1–130)....... positive isolates of the outbreak strain had been found in a particular pig slaughterhouse and thus early traceback, investigation and control measures were possible. Several batches of pork and pork products were recalled and the slaughterhouse was closed twice for disinfection. No single common food item...

Effects of chemical and physical mutagens on the frequency of a large genetic duplication in Salmonella typhimurium. I

International Nuclear Information System (INIS)

Hoffman, G.R.; Morgan, R.W.

1978-01-01

In Salmonella typhimurium a simple selection has been described to detect bacteria that are merodiploid for almost one-third of the chromosome. The selective procedure is based upon improved utilization of L-malate as the sole carbon source in merodiploid strains. The spontaneous frequency of the duplication in haploid strains is approximately 10 -4 per cell plated. Following the exposure of a haploid strain to mutagenic agents, there is a dose-dependent increase in the duplication frequency above the spontaneous level. In this paper the authors describe the induction of genetic duplications in Salmonella typhimurium by X-rays, ultraviolet light (UV), ethyl methanesulfonate (EMS), nitrous acid, and the azaacridine half mustard, ICR-372. (Auth.)

Effect of wine-based marinades on the behavior of Salmonella Typhimurium and background flora in beef fillets.

Science.gov (United States)

Nisiotou, A; Chorianopoulos, N G; Gounadaki, A; Panagou, E Z; Nychas, G-J E

2013-06-17

The aim of this study was to evaluate the wine-based marinades to control the survival of acid-adapted and non-adapted Salmonella Typhimurium and background flora of fresh beef stored aerobically or under modified atmosphere. Beef slices were inoculated with a 3-strain cocktail of acid-adapted or non-adapted Salmonella Typhimurium strains DT 193, 4/74 and DSM 554 and marinated by immersion in wine (W) or wine supplemented with 0.3% thyme essential oil (WEO), for 12h at 4°C. Marinated slices were then stored under air or modified atmosphere conditions at 5°C. S. Typhimurium and background flora were followed for a 19-day period of storage. S. Typhimurium individual strains were monitored by pulsed field gel electrophoresis. Marination with wine significantly (Pwine marinade. The epidemic multi-drug resistant DT 193, the 4/74 and DSM 554 strains survived marination (for both W and WEO) and were detected at about similar proportions as revealed by PFGE results. Present results indicate that wine-based marinades are efficient, from a safety and shelf life stand point, in reducing pathogen's levels as well as the background beef flora. Copyright © 2013 Elsevier B.V. All rights reserved.

A new wireless detection device for the in-situ identification of Salmonella Typhimurium

Science.gov (United States)

Chai, Yating; Wikle, Howard C.; Park, Mi-kyung; Horikawa, Shin; Hong, Xie; Chin, Bryan A.

2013-05-01

This paper presents a new device and method for the in-situ detection of Salmonella Typhimurium on tomato surfaces. This real-time in-situ detection was accomplished with phage-based magnetoelastic (ME) biosensors on fresh food surfaces. The E2 phage from a landscape phage library serves as the bio-recognition element that has the capability of binding specifically with S. Typhimurium. This mass-sensitive ME biosensor is wirelessly actuated into mechanical resonance by an externally applied time-varying magnetic field. When the biosensor binds with S. Typhimurium, the mass of the sensor increases, resulting in a decrease in the sensor's resonant frequency. Until now, ME sensors had to be collected from the tomato surface where they are exposed to S. Typhimurium and inserted into a measurement coil for the detection of the bacterium. In contrast, the newly designed test device allows the whole detection process to take place directly on the tomato. Changes in resonant frequency over time due to the accumulation of S. Typhimurium on the sensor were measured and are presented. Real-time in-situ detection of 20 minutes was achieved. In addition, this new methodology effectively decreases the measurement error and enables the simultaneous detection of multiple pathogens.

Behaviour of Salmonella Typhimurium during production and storage of artisan water buffalo mozzarella cheese

Directory of Open Access Journals (Sweden)

Roberto Rosmini

2012-07-01

Full Text Available Water buffalo mozzarella cheese (WBMC is a fresh pasta filata cheese produced from whole chilled buffalo milk. Although pasteurization of milk and the use of defined starter cultures are recommended, traditional technology involving the use of unpasteurized milk and natural whey cultures is still employed for WBMC production in Italy. The aim of this study were to assess the behaviour of Salmonella Typhimurium during the production of artisan water buffalo mozzarella cheese and during its shelf life under different temperature conditions. Raw milk was inoculated with S. Typhimurium and the evolution of S. Typhimurium count during production and shelf life was monitored. In artisan WBMC production technology S. Typhimurium multiplied in the curd during ripening, but its growth rate expressed in log CFU/g/h was lower than the growth rate reported by theoretical predictions. Stretching proved to be a process with good repeatability and able to reduce S. Typhimurium contamination by 5.5 Log CFU/g. The intrinsic characteristics of traditional WBMC proved to be unable to obstacolate the growth of S. Typhimurium during storage in the case of thermal abuse. Control of raw milk contamination and a proper refrigeration temperature are key factors in reducing the risk for consumers.

Observations on the distribution and control of Salmonella in commercial duck hatcheries in the UK.

Science.gov (United States)

Martelli, F; Birch, C; Davies, R H

2016-01-01

Salmonella infection causes a significant number of cases of gastroenteritis and more serious illnesses in people in the UK and EU. The serovars Salmonella Enteritidis and Salmonella Typhimurium are most frequently associated with foodborne illness in Europe. Whilst control programmes exist to monitor these serovars in the chicken and turkey sectors, no regulatory programme is currently in place for the duck sector. A voluntary industry scheme (Duck Assurance Scheme) was launched in the UK in 2010. Hatcheries act as focal points of Salmonella contamination, in particular if Salmonella-contaminated eggs from positive breeding farms enter the hatchery. Five duck hatcheries were visited in this study and four were positive for Salmonella. S. Typhimurium DT8 and S. Indiana were isolated from hatchery 1 and S. Typhimurium DT41 and S. Senftenberg were isolated from hatchery 3. S. Kottbus, S. Bovismorbificans and S. Senftenberg were isolated from hatchery 2 and S. Kedougou was isolated from hatchery 4. Advice on the control/elimination of Salmonella was provided at each visit and a longitudinal study was undertaken to monitor its effectiveness. Extensive sampling was carried out in the hatcheries visited and the tray wash area and waste/external areas had the highest probability of being contaminated. The hatcher area was also found to be a primary focus of contamination. Improvements of farm and hatchery biosecurity standards have resulted in a reduction of hatchery contamination in this study and in previous investigations. Hatcheries 1 and 5 were cleared of Salmonella, demonstrating that elimination of Salmonella contamination from duck hatcheries is achievable.

The occurrence of Salmonella spp. in duck eggs on sale at retail or from catering in England.

Science.gov (United States)

Owen, M; Jorgensen, F; Willis, C; McLauchlin, J; Elviss, N; Aird, H; Fox, A; Kaye, M; Lane, C; de Pinna, E

2016-11-01

Since 2010, human salmonellosis outbreaks in the UK have been detected as associated with the consumption of duck eggs. Little data are available on the rate of occurrence of Salmonella in duck eggs. The aim of this study was to investigate the occurrence of Salmonella spp. in duck eggs on sale and from catering in England during 2011, particularly those from small-scale production. All samples were collected independently of human salmonellosis outbreak investigations. Composite samples of 6-10 eggs (shells and contents were examined separately) were examined for the presence of Salmonella spp. using the ISO 6579:2002 method. Salmonella spp. was recovered from two of 145 samples (1·4%). In one sample, Salmonella Typhimurium DT 8 was isolated from the shells while Salm. Typhimurium DT 8 and Salm. Typhimurium DT30 were isolated from the contents. Salmonella Typhimurium DT8 was isolated from the egg shells only in the second contaminated sample. This study provides baseline data for risk assessors, regulators and the food industry and may be helpful in communicating risks associated with the consumption of this product as well as evaluating risk management options to control food safety including vaccination of ducks. Human salmonellosis outbreaks in England and Northern Ireland due to Salmonella enterica serovar Typhimurium definitive phage type (DT) 8 have been identified as associated with the consumption of duck eggs since 2010. This study has shown that Salmonella spp. was detected in 1·4% of ducks egg samples providing baseline data for risk assessors, regulators and the food industry. This may be helpful in communicating risks associated with the consumption of this product as well as evaluating risk management options to control food safety including vaccination of ducks. © 2016 Crown copyright. Letters in Applied Microbiology © 2016 The Society for Applied Microbiology.

Co-ordinate regulation of Salmonella typhimurium invasion genes by environmental and regulatory factors is mediated by control of hilA expression.

Science.gov (United States)

Bajaj, V; Lucas, R L; Hwang, C; Lee, C A

1996-11-01

During infection of their hosts, salmonellae enter intestinal epithelial cells. It has been proposed that when Salmonella typhimurium is present in the intestinal lumen, several environmental and regulatory conditions modulate the expression of invasion factors required for bacterial entry into host cells. We report here that the expression of six different S. typhimurium invasion genes encoded on SPI1 (Salmonella pathogenicity island 1) is co-ordinately regulated by oxygen, osmolarity, pH, PhoPQ, and HilA. HilA is a transcriptional activator of the OmpR/ToxR family that is also encoded on SPI1. We have found that HilA plays a central role in the co-ordinated regulation of invasion genes by environmental and regulatory conditions. HilA can activate the expression of two invasion gene-lacZY fusions on reporter plasmids in Escherichia coll, suggesting that HilA acts directly at invasion-gene promoters in S. typhimurium. We have found that the regulation of invasion genes by oxygen, osmolarity, pH, and PhoPQ is indirect and is mediated by regulation of hilA expression by these environmental and regulatory factors. We hypothesize that the complex and co-ordinate regulation of Invasion genes by HilA is an important feature of salmonella pathogenesis and allows salmonellae to enter intestinal epithelial cells.

Long-term kinetics of Salmonella Typhimurium ATCC 14028 survival on peanuts and peanut confectionery products.

Directory of Open Access Journals (Sweden)

Maristela S Nascimento

Full Text Available Due to recent large outbreaks, peanuts have been considered a product of potential risk for Salmonella. Usually, peanut products show a low water activity (aw and high fat content, which contribute to increasing the thermal resistance and survival of Salmonella. This study evaluated the long-term kinetics of Salmonella survival on different peanut products under storage at 28°C for 420 days. Samples of raw in-shell peanuts (aw = 0.29, roasted peanuts (aw = 0.39, unblanched peanut kernel (aw = 0.54, peanut brittle (aw = 0.30, paçoca (aw = 0.40 and pé-de-moça (aw = 0.68 were inoculated with Salmonella Typhimurium ATCC 14028 at two inoculum levels (3 and 6 log cfu/ g. The Salmonella behavior was influenced (p420 days, especially in products with aw around 0.40.

Prevalence, Virulence Genes and Antimicrobial Resistance Profiles of Salmonella Serovars from Retail Beef in Selangor, Malaysia.

Science.gov (United States)

Thung, Tze Y; Radu, Son; Mahyudin, Nor A; Rukayadi, Yaya; Zakaria, Zunita; Mazlan, Nurzafirah; Tan, Boon H; Lee, Epeng; Yeoh, Soo L; Chin, Yih Z; Tan, Chia W; Kuan, Chee H; Basri, Dayang F; Wan Mohamed Radzi, Che W J

2017-01-01

The aim of the present study was to investigate the prevalence of Salmonella spp., Salmonella Enteritidis and Salmonella Typhimurium in retail beef from different retail markets of Selangor area, as well as, to assess their pathogenic potential and antimicrobial resistance. A total of 240 retail beef meat samples (chuck = 60; rib = 60; round = 60; sirloin = 60) were randomly collected. The multiplex polymerase chain reaction (mPCR) in combination with the most probable number (MPN) method was employed to detect Salmonella spp., S . Enteritidis and S . Typhimurium in the meat samples. The prevalence of Salmonella spp., S . Enteritidis and S . Typhimurium in 240 beef meat samples were 7.50, 1.25, and 0.83%, respectively. The microbial loads of total Salmonella was found in the range of retail beef products tested were widely contaminated with multi-drug resistant (MDR) Salmonella and various virulence genes are present among the isolated Salmonella serovars.

Microbial radio-resistance of Salmonella Typhimurium in egg increases due to repetitive irradiation with electron beam

International Nuclear Information System (INIS)

Tesfai, Adiam T.; Beamer, Sarah K.; Matak, Kristen E.; Jaczynski, Jacek

2011-01-01

Ionizing radiation improves food safety. However, foodborne pathogens develop increased resistance in response to sub-lethal stresses such as heat, pH, antibiotics, etc. Therefore, it is hypothesized that foodborne pathogens may develop increased radio-resistance to electron beam (e-beam) radiation. The objective was to determine if D 10 -value for Salmonella Typhimurium in de-shelled raw egg (egg white and yolk mixed together) increases due to repetitive processing with e-beam at sub-lethal doses. Survivors were enumerated on non-selective (TSA) and selective (XLD) media. Survivors from the highest dose were isolated and used in subsequent e-beam cycle. This process was repeated four times for a total of five e-beam cycles. D 10 -values for S. Typhimurium enumerated on TSA and XLD following each e-beam cycle were calculated as inverse reciprocal of the slope of survivor curves. D 10 -values for the ATCC strain were 0.59±0.031 and 0.46±0.022 kGy on TSA and XLD, respectively. However, following the fifth e-beam cycle, the respective D 10 -values increased (P 0.05) to develop radio-resistance faster on selective media, likely due to facilitated selection of radio-resistant cells within microbial population following each e-beam cycle. For all five e-beam cycles, S. Typhimurium had higher (P 10 -values on non-selective media, indicating that sub-lethal injury followed by cellular repair and recovery are important for radio-resistance and inactivation of this microorganism. This study demonstrated that e-beam efficiently inactivates S. Typhimurium in raw egg; however, similar to other inactivation techniques and factors affecting microbial growth, S. Typhimurium develops increased radio-resistance if repetitively processed with e-beam at sub-lethal doses.

Phenotypic and Genotypic Eligible Methods for Salmonella Typhimurium Source Tracking.

Science.gov (United States)

Ferrari, Rafaela G; Panzenhagen, Pedro H N; Conte-Junior, Carlos A

2017-01-01

Salmonellosis is one of the most common causes of foodborne infection and a leading cause of human gastroenteritis. Throughout the last decade, Salmonella enterica serotype Typhimurium (ST) has shown an increase report with the simultaneous emergence of multidrug-resistant isolates, as phage type DT104. Therefore, to successfully control this microorganism, it is important to attribute salmonellosis to the exact source. Studies of Salmonella source attribution have been performed to determine the main food/food-production animals involved, toward which, control efforts should be correctly directed. Hence, the election of a ST subtyping method depends on the particular problem that efforts must be directed, the resources and the data available. Generally, before choosing a molecular subtyping, phenotyping approaches such as serotyping, phage typing, and antimicrobial resistance profiling are implemented as a screening of an investigation, and the results are computed using frequency-matching models (i.e., Dutch, Hald and Asymmetric Island models). Actually, due to the advancement of molecular tools as PFGE, MLVA, MLST, CRISPR, and WGS more precise results have been obtained, but even with these technologies, there are still gaps to be elucidated. To address this issue, an important question needs to be answered: what are the currently suitable subtyping methods to source attribute ST. This review presents the most frequently applied subtyping methods used to characterize ST, analyses the major available microbial subtyping attribution models and ponders the use of conventional phenotyping methods, as well as, the most applied genotypic tools in the context of their potential applicability to investigates ST source tracking.

Phenotypic and Genotypic Eligible Methods for Salmonella Typhimurium Source Tracking

Directory of Open Access Journals (Sweden)

Rafaela G. Ferrari

2017-12-01

Full Text Available Salmonellosis is one of the most common causes of foodborne infection and a leading cause of human gastroenteritis. Throughout the last decade, Salmonella enterica serotype Typhimurium (ST has shown an increase report with the simultaneous emergence of multidrug-resistant isolates, as phage type DT104. Therefore, to successfully control this microorganism, it is important to attribute salmonellosis to the exact source. Studies of Salmonella source attribution have been performed to determine the main food/food-production animals involved, toward which, control efforts should be correctly directed. Hence, the election of a ST subtyping method depends on the particular problem that efforts must be directed, the resources and the data available. Generally, before choosing a molecular subtyping, phenotyping approaches such as serotyping, phage typing, and antimicrobial resistance profiling are implemented as a screening of an investigation, and the results are computed using frequency-matching models (i.e., Dutch, Hald and Asymmetric Island models. Actually, due to the advancement of molecular tools as PFGE, MLVA, MLST, CRISPR, and WGS more precise results have been obtained, but even with these technologies, there are still gaps to be elucidated. To address this issue, an important question needs to be answered: what are the currently suitable subtyping methods to source attribute ST. This review presents the most frequently applied subtyping methods used to characterize ST, analyses the major available microbial subtyping attribution models and ponders the use of conventional phenotyping methods, as well as, the most applied genotypic tools in the context of their potential applicability to investigates ST source tracking.

Antimicrobial susceptibility profile, treatment outcome and serotype distribution of clinical isolates of Salmonella enterica subspecies enterica: a 2-year study from Kerala, South India

Directory of Open Access Journals (Sweden)

Harichandran D

2017-03-01

Full Text Available Deepa Harichandran, Kavitha Radhakrishnan Dinesh Department of Microbiology, Amrita School of Medicine, Amrita Institute of Medical Sciences and Research Centre, Amrita Vishwa Vidyapeetham University, Kochi, Kerala, India Background/purpose: Typhoid and paratyphoid fever continue to be important causes of illness and death in parts of Asia, being associated with poor sanitation and consumption of unsafe food and water. Antimicrobial resistance has emerged to traditional first-line drugs, namely, the fluoroquinolones, as well as to third-generation cephalosporins, posing challenges to treatment. Azithromycin has proven to be an effective alternative for treatment of uncomplicated typhoid fever. The purpose of this study was to determine the antimicrobial susceptibility, clinical outcome and serotype distribution pattern of clinical isolates belonging to Salmonella enterica subspecies enterica. Methodology: All clinical isolates of S. enterica obtained from blood, sterile body fluids, as well as stool and urine samples at Amrita Institute of Medical Sciences and Research Centre, Kerala, India, between August 2011 and July 2013 were included in the study and processed based on standard microbiology protocols. Results: A total of 118 isolates of Salmonella were obtained during the study period. Out of these, 79 were of S. Typhi (66.95%, followed by isolates of S. Paratyphi A (22; 18.64% and S. Typhimurium 12 (10.17%. Five isolates could not be identified further. There was 100% susceptibility to ceftriaxone in all S. enterica subspecies. Ciprofloxacin susceptibility was 32.91% for S. Typhi and 40.90% for S. Paratyphi A as determined by the disk diffusion method. The susceptibility profile of S. Typhi isolates to different antimicrobials was as follows: chloramphenicol (94.93%, ampicillin (77.21%, cotrimoxazole (75.94% and azithromycin (78.48%. For S. Typhi, the minimum inhibitory concentration (MIC of ciprofloxacin required to inhibit the growth

Molecular typing, antibiotic resistance, virulence gene and biofilm formation of different Salmonella enterica serotypes.

Science.gov (United States)

Turki, Yousra; Mehr, Ines; Ouzari, Hadda; Khessairi, Amel; Hassen, Abdennaceur

2014-01-01

Salmonella enterica isolates representing commonly isolated serotypes in Tunisia were analyzed using genotyping and phenotyping methods. ERIC and ITS-PCR applied to 48 Salmonella spp. isolates revealed the presence of 12 and 10 different profiles, respectively. The distribution of profiles among serotypes demonstrated the presence of strains showing an identical fingerprinting pattern. All Salmonella strains used in this study were positive for the sdiA gene. Three Salmonella isolates belonging to serotypes Anatum, Enteritidis and Amsterdam were negative for the invA gene. The spvC gene was detected in thirteen isolates belonging to serotypes Anatum, Typhimurium, Enteritidis, Gallinarum and Montevideo. Antibiotic resistance was frequent among the recovered Salmonella isolates belonging to serotypes Anatum, Typhimurium, Enteritidis, Zanzibar and Derby. The majority of these isolates exhibited resistance to at least two antibiotic families. Four multidrug-resistant isolates were recovered from food animals and poultry products. These isolates exhibited not only resistance to tetracycline, sulphonamides, and ampicillin, but also have shown resistance to fluoroquinolones. Common resistance to nalidixic acid, ciprofloxacin and ofloxacin in two S. Anatum and S. Zanzibar strains isolated from raw meat and poultry was also obtained. Furthermore, wastewater and human isolates exhibited frequent resistance to nalidixic acid and tetracycline. Of all isolates, 33.5% were able to form biofilm.

Quinolone Resistance among Salmonella enterica from Cattle, Broilers and Swine in Denmark

DEFF Research Database (Denmark)

Wiuff, C.; Baggesen, Dorte Lau; Madsen, M.

2000-01-01

This study was conducted to determine the susceptibility to nalidixic acid and fluoroquinolones of Salmonella Dublin, S. Enteritidis, and S. Typhimurium isolates from cattle, broilers, and pigs over time in Denmark and to characterise the gyrA, gyrB, and parC genes in quinolone-resistant isolates...... that quinolone-resistant isolates have emerged in recent years among food-producing animals, especially among S. Enteritidis from broilers in Denmark, and that the resistance mainly is associated with mutations in gyrA.......This study was conducted to determine the susceptibility to nalidixic acid and fluoroquinolones of Salmonella Dublin, S. Enteritidis, and S. Typhimurium isolates from cattle, broilers, and pigs over time in Denmark and to characterise the gyrA, gyrB, and parC genes in quinolone-resistant isolates...... to quinolones. A single (1.1%) S. Typhimurium isolate from 1995 and three (5.9%) from 1998 were resistant to nalidixic acid. Six (9.0%) S. Dublin isolates from 1996, four (4.2%) from 1997, and one (1.7%) from 1998 were resistant to nalidixic acid. Resistance was not observed among isolates from cattle in 1999...

Contribution of different mechanisms to the resistance to fluoroquinolones in clinical isolates of Salmonella enterica

Directory of Open Access Journals (Sweden)

Abeer Ahmed Rushdy

Full Text Available OBJECTIVES: To study the potential factors include gene mutation, efflux pump and alteration of permeability associated with quinolone-resistance of Salmonella enterica strains isolated from patients with acute gastroenteritis and to evaluate the degree of synergistic activity of efflux pump inhibitors when combined with ciprofloxacin against resistant isolates. METHODS: Antimicrobial resistance patterns of fifty-eight Salmonella isolates were tested. Five isolates were selected to study the mechanism of resistance associated with quinolone group, including mutation in topoisomerase-encoding gene, altered cell permeability, and expression of an active efflux system. In addition, the combination between antibiotics and efflux pump inhibitors to overcome the microbial resistance was evaluated. RESULTS: Five Salmonella isolates totally resistant to all quinolones were studied. All isolates showed alterations in outer membrane proteins including disappearance of some or all of these proteins (Omp-A, Omp-C, Omp-D and Omp-F. Minimum inhibitory concentration values of ciprofloxacin were determined in the presence/absence of the efflux pump inhibitors: carbonyl cyanide m-chlorophenylhydrazone, norepinephrin and trimethoprim. Minimum inhibitory concentration values for two of the isolates were 2-4 fold lower with the addition of efflux pump inhibitors. All five Salmonella isolates were amplified for gyrA and parC genes and only two isolates were sequenced. S. Enteritidis 22 had double mutations at codon 83 and 87 in addition to three mutations at parC at codons 67, 76 and 80 whereas S. Typhimurium 57 had three mutations at codons 83, 87 and 119, but no mutations at parC. CONCLUSIONS: Efflux pump inhibitors may inhibit the major AcrAB-TolC in Salmonella efflux systems which are the major efflux pumps responsible for multidrug resistance in Gramnegative clinical isolates.

Evaluation of the addition of organic acids in the feed and/or water for broilers and the subsequent recovery of Salmonella Typhimurium from litter and ceca.

Science.gov (United States)

Bourassa, D V; Wilson, K M; Ritz, C R; Kiepper, B K; Buhr, R J

2018-01-01

Three separate broiler Salmonella Typhimurium challenge experiments were conducted evaluating efficacy of formic and propionic acid feed supplements to suppress environmental and cecal Salmonella Typhimurium prevalence. In experiment 1, broilers were provided feed with 1 kg/ton formic acid or 5 kg/ton propionic acid feed additives or a basal control diet. At the day of placement, half of the pens were inoculated with seeder chicks orally challenged with a marker strain of Salmonella Typhimurium and to yield challenged and adjacent nonchallenged pens. No differences in weekly litter samples or cecal Salmonella prevalence at 3 or 6 wk among feeding treatments were detected. In experiment 2, treatments were: 2 kg/ton propionic acid in feed, 1.0 mL/L formic acid in water, both propionic acid in feed and formic acid in water, and a basal control. Every pen was challenged with seeder chicks inoculated with Salmonella Typhimurium. By 6 wk all pens maintained detectable litter Salmonella, and broilers provided both propionic acid in feed and formic acid in water had the lowest cecal recovery (35%), compared to the control (60%). In experiment 3, treatments were: formic acid at 4 or 6 kg/ton from wk 0 to 6 or for only the last wk, propionic acid at 5 or 10 kg/ton for only the last wk, and a basal control. Each pen was challenged with Salmonella Typhimurium inoculated seeder chicks. By 6 wk, broilers fed formic acid (4 kg/ton) for the entire growout had no Salmonella-positive ceca (0/30). All treatments that provided acid supplemented feed for only the last wk had 3-13% Salmonella-positive ceca. These experiments indicate that adding formic acid to broiler feed appears to prevent Salmonella colonization from challenge pens entering into the adjacent nonchallenge pens. Feeding formic acid (4 kg/ton) for 6 wk resulted in no recovery of Salmonella from ceca compared to the control prevalence of 17%. Published by Oxford University Press on behalf of Poultry Science

Impact of relative humidity, inoculum carrier and size, and native microbiota on Salmonella ser. Typhimurium survival in baby lettuce.

Science.gov (United States)

López-Gálvez, Francisco; Gil, Maria Isabel; Allende, Ana

2018-04-01

The effects of relative humidity (RH), fluctuating climate conditions, inoculum size and carrier on the survival of Salmonella enterica serovar Typhimurium on baby lettuce in environmental test chambers were studied. Buffered peptone water (BPW), distilled water (DW), and irrigation water (IW) were compared as inoculum carriers. Additionally, survival of Salmonella in suspensions prepared using filtered and unfiltered IW was assessed. Salmonella Typhimurium survived better on baby lettuce plants at high RH independently of the inoculum size. When lettuce plants were grown under fluctuating environmental conditions, Salmonella survival was similar under both RH conditions. Regarding the inoculum carrier, the inoculated microorganism survived better on lettuce plants when BPW was used as carrier both at high and low RH. Survival rate of Salmonella in IW was affected by the presence of native microbiota. Native microbiota present in IW did not affect survival of Salmonella or the levels of mesophilic bacteria on the baby lettuce leaves. The information obtained in the present study contributes to the knowledge on the effect of environmental conditions on pathogenic bacteria survival on growing edible plants. These results are useful when selecting the methodology to carry out experimental studies on the survival of microbial pathogens under different pre-harvest conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

Managing Salmonella Typhimurium and Escherichia coli O157:H7 in soil with hydrated lime - An outdoor study in lysimeters and field plots.

Science.gov (United States)

Nyberg, Karin A; Vinnerås, Björn; Albihn, Ann

2014-01-01

An outbreak of Salmonella Typhimurium or E. coli O157:H7 among domestic animals can have great financial consequences for an animal enterprise but also be a threat for public health as there is a risk for transmission of the infection through the environment. In order to minimize disease transmission, it is important to treat not only the affected animals but also the areas on which they have been kept. In the present study, the effect of hydrated lime as a treatment for Salmonella Typhimurium or E. coli O157:H7 contaminated soil was investigated. The study was performed outdoors, in a lysimeter system and in field plots. The soils were spiked with Salmonella Typhimurium and/or E. coli O157:H7 and hydrated lime was added at three different concentrations (0.5, 1 and 2%). Sampling was performed over one month, and the levels of bacteria were analyzed by standard culture methods. In addition, the soil pH was monitored throughout the study. The results showed that application of 0.5-1 kg hydrated lime per m(2) reduced both Salmonella Typhimurium and E. coli O157:H7 numbers to below the detection limit (2 log10 CFU g-1 soil) in 3-7 days. Lower application rates of hydrated lime did not reduce pathogen numbers in the lysimeter study, but in the field plots no E. coli O157:H7 was detected at the end of the four-week study period regardless of hydrated lime application. A recommended strategy for treating a Salmonella Typhimurium or E. coli O157:H7 contaminated soil could therefore be to monitor the pH over the time of treatment and to repeat hydrated lime application if a decrease in pH is observed.

Branched-chain amino acid supplementation promotes aerobic growth of Salmonella Typhimurium under nitrosative stress conditions.

Science.gov (United States)

Park, Yoon Mee; Lee, Hwa Jeong; Jeong, Jae-Ho; Kook, Joong-Ki; Choy, Hyon E; Hahn, Tae-Wook; Bang, Iel Soo

2015-12-01

Nitric oxide (NO) inactivates iron-sulfur enzymes in bacterial amino acid biosynthetic pathways, causing amino acid auxotrophy. We demonstrate that exogenous supplementation with branched-chain amino acids (BCAA) can restore the NO resistance of hmp mutant Salmonella Typhimurium lacking principal NO-metabolizing enzyme flavohemoglobin, and of mutants further lacking iron-sulfur enzymes dihydroxy-acid dehydratase (IlvD) and isopropylmalate isomerase (LeuCD) that are essential for BCAA biosynthesis, in an oxygen-dependent manner. BCAA supplementation did not affect the NO consumption rate of S. Typhimurium, suggesting the BCAA-promoted NO resistance independent of NO metabolism. BCAA supplementation also induced intracellular survival of ilvD and leuCD mutants at wild-type levels inside RAW 264.7 macrophages that produce constant amounts of NO regardless of varied supplemental BCAA concentrations. Our results suggest that the NO-induced BCAA auxotrophy of Salmonella, due to inactivation of iron-sulfur enzymes for BCAA biosynthesis, could be rescued by bacterial taking up exogenous BCAA available in oxic environments.

Effects of bacterial dose and fly sex on persistence and excretion of Salmonella enterica serovar Typhimurium from adult house flies (Diptera: Muscidae)

Science.gov (United States)

Salmonella Typhimurium is a pathogen that causes gastroenteritis in humans and can be harbored by house flies. Factors influencing excretion of S. Typhimurium from infected flies have not been elucidated, but are essential for assessing transmission potential. We determined the persistence and excre...

Antibacterial effect of roselle extracts (Hibiscus sabadariffa), sodium hypochlorite and acetic acid against multidrug-resistant Salmonella strains isolated from tomatoes.

Science.gov (United States)

Gutiérrez-Alcántara, E J; Rangel-Vargas, E; Gómez-Aldapa, C A; Falfan-Cortes, R N; Rodríguez-Marín, M L; Godínez-Oviedo, A; Cortes-López, H; Castro-Rosas, J

2016-02-01

Antibiotic-resistant Salmonella strains were isolated from saladette and red round type tomatoes, and an analysis done of the antibacterial activity of roselle calyx extracts against any of the identified strains. One hundred saladette tomato samples and 100 red round tomato samples were collected from public markets. Each sample consisted of four whole tomatoes. Salmonella was isolated from the samples by conventional culture procedure. Susceptibility to 16 antibiotics was tested for the isolated Salmonella strains by standard test. The antibacterial effect of four roselle calyx extracts (water, methanol, acetone and ethyl acetate), sodium hypochlorite and acetic acid against antibiotic-resistant Salmonella isolates was evaluated on contaminated tomatoes. Twenty-four Salmonella strains were isolated from 12% of each tomato type. Identified Salmonella serotypes were Typhimurium and Typhi. All isolated strains exhibited resistance to at least three antibiotics and some to as many as 12. Over contaminated tomatoes, the roselle calyx extracts produced a greater reduction (2-2·6 log) in antibiotic-resistant Salmonella strain concentration than sodium hypochlorite and acetic acid. The presence of multidrug-resistant Salmonella in vegetables is a significant public health concern. Multidrug-resistant Salmonella strains were isolated from raw tomatoes purchased in public markets in Mexico and challenged with roselle Hibiscus sabdariffa calyx extracts, sodium hypochlorite and acetic acid. On tomatoes, the extracts caused a greater reduction in the concentration of antibiotic-resistant Salmonella strains than sodium hypochlorite and acetic acid. Roselle calyx extracts are a potentially useful addition to disinfection procedures of raw tomatoes in the field, processing plants, restaurants and homes. © 2015 The Society for Applied Microbiology.

Transfer of primed CD4+OX40- T lymphocytes induces increased immunity to experimental Salmonella typhimurium infections in rats

DEFF Research Database (Denmark)

Thygesen, P; Christensen, H B; Hougen, H P

1997-01-01

The protective effect of primed CD4 T cells against a lethal dose of Salmonella typhimurium was studied in Lewis rats. Primed CD4 T cells were obtained by inoculating Lewis rats with a non-lethal dose of S. typhimurium. Four weeks after the infection, spleen non-adherent mononuclear cells were is......-specific memory T cells that have returned to a resting state....

Generalised linear mixed models analysis of risk factors for contamination of Danish broiler flocks with Salmonella typhimurium

DEFF Research Database (Denmark)

Chriél, Mariann; Stryhn, H.; Dauphin, G.

1999-01-01

are the broiler flocks (about 4000 flocks) which are clustered within producers. Broiler flocks with ST-infected parent stocks show increased risk of salmonella infection, and also the hatchery affects the salmonella status significantly. Among the rearing factors, only the use of medicine as well as the time......We present a retrospective observational study of risk factors associated with the occurrence of Salmonella typhimurium (ST) in Danish broiler flocks. The study is based on recordings from 1994 in the ante-mortem database maintained by the Danish Poultry Council. The epidemiological units...

Salmonella typhimurium infection in total knee arthroplasty: A case report with review of literature

Directory of Open Access Journals (Sweden)

Sujeesh Sebastian

2017-01-01

Full Text Available Salmonella enterica serotype Typhimurium is a rare cause of prosthetic joint infection (PJI. The recognized predisposing risk factors for Salmonella septic arthritis include diabetes mellitus, renal failure, human immunodeficiency virus infection and chronic corticosteroid use. We describe a case of PJI of the knee in a 74-year-old lady who was on antitubercular treatment. The patient presented with discharging sinus and raised inflammatory markers. She was successfully treated by the removal of prosthesis and debridement followed by ciprofloxacin therapy for 6 weeks. This case report highlights the potential virulence of Salmonella in immunocompromised patient with a joint prosthesis. Continuous monitoring and close collaboration of microbiologists and orthopedicians helped obtain the resolution of infection in our patient.

Spray method for recovery of heat-injured Salmonella Typhimurium and Listeria monocytogenes.

Science.gov (United States)

Back, Kyeong-Hwan; Kim, Sang-Oh; Park, Ki-Hwan; Chung, Myung-Sub; Kang, Dong-Hyun

2012-10-01

Selective agar is inadequate for supporting recovery of injured cells. During risk assessment of certain foods, both injured and noninjured cells must be enumerated. In this study, a new method (agar spray method) for recovering sublethally heat-injured microorganisms was developed and used for recovery of heat-injured Salmonella Typhimurium and Listeria monocytogenes. Molten selective agar was applied as an overlay to presolidified nonselective tryptic soy agar (TSA) by spray application. Heat-injured cells (55°C for 10 min in 0.1% peptone water or 55°C for 15 min in sterilized skim milk) were inoculated directly onto solidified TSA. After a 2-h incubation period for cell repair, selective agar was applied to the TSA surface with a sprayer, and the plates were incubated. The recovery rate for heat-injured Salmonella Typhimurium and L. monocytogenes with the spray method was compared with the corresponding rates associated with TSA alone, selective media alone, and the conventional overlay method (selective agar poured on top of resuscitated cells grown on TSA and incubated for 2 h). No significant differences (P > 0.05) were found in pathogen recovery obtained with TSA, the overlay method, and the spray method. However, a lower recovery rate (P recovery and detection of injured cells.

A method for investigating protein-protein interactions related to Salmonella typhimurium pathogenesis

Energy Technology Data Exchange (ETDEWEB)

Chowdhury, Saiful M. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Shi, Liang [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Yoon, Hyunjin [Dartmouth College, Hanover, NH (United States); Ansong, Charles [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Rommereim, Leah M. [Dartmouth College, Hanover, NH (United States); Norbeck, Angela D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Auberry, Kenneth J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Moore, R. J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Adkins, Joshua N. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Heffron, Fred [Oregon Health and Science Univ., Portland, OR (United States); Smith, Richard D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

2009-02-10

We successfully modified an existing method to investigate protein-protein interactions in the pathogenic bacterium Salmonella typhimurium (STM). This method includes i) addition of a histidine-biotin-histidine tag to the bait proteins via recombinant DNA techniques; ii) in vivo cross-linking with formaldehyde; iii) tandem affinity purification of bait proteins under fully denaturing conditions; and iv) identification of the proteins cross-linked to the bait proteins by liquid-chromatography in conjunction with tandem mass-spectrometry. In vivo cross-linking stabilized protein interactions permitted the subsequent two-step purification step conducted under denaturing conditions. The two-step purification greatly reduced nonspecific binding of non-cross-linked proteins to bait proteins. Two different negative controls were employed to reduce false-positive identification. In an initial demonstration of this approach, we tagged three selected STM proteins- HimD, PduB and PhoP- with known binding partners that ranged from stable (e.g., HimD) to transient (i.e., PhoP). Distinct sets of interacting proteins were identified with each bait protein, including the known binding partners such as HimA for HimD, as well as anticipated and unexpected binding partners. Our results suggest that novel protein-protein interactions may be critical to pathogenesis by Salmonella typhimurium. .

Experimental Salmonella typhimurium infections in rats. I

DEFF Research Database (Denmark)

Hougen, H P; Jensen, E T; Klausen, B

1989-01-01

The course of experimentally induced Salmonella typhimurium infection was studied in three groups of inbred LEW rats: homozygous +/+, athymic rnu/rnu and isogeneic thymus-grafted rnu/rnu rats. In the first experiment the animals were inoculated intraperitoneally with 10(8) bacteria and all animals...... became severely septicemic and died within a week of inoculation, irrespective of presence or absence of thymus. In the second experiment the animals were inoculated with 10(6) bacteria, and both euthymic and thymus-grafted animals responded with high titres of anti bacterial antibodies while these were...... very low in the athymic nude animals. Polyclonal antibody production was only observed in the euthymic animals and only regarding IgG. Athymic rats were not able to clear the infection, while the thymus-grafted animals reacted like euthymic rats: Very few animals housed the bacteria four weeks after...

Utilization of a novel autologous killed tri-vaccine (serogroups B [Typhimurium], C [Mbandaka] and E [Orion]) for Salmonella control in commercial poultry breeders.

Science.gov (United States)

Pavic, Anthony; Groves, Peter J; Cox, Julian M

2010-02-01

An autologous killed trivalent vaccine (3x10(8) colony-forming units [CFU]), based on three Salmonella serovars (Typhimurium - serogroup B, Mbandaka - serogroup C, and Orion - serogroup E) prevalent in the flocks of Australian poultry companies, was developed using Salenvac techniques. At 20 weeks, hens vaccinated at 12 and 17 weeks as well as non-vaccinated hens were challenged (250 microl of 10(7) CFU) with autologous and heterologous serovars belonging to serogroup B (Typhimurium and Agona), serogroup C (Mbandaka and Infantis) and serogroup E (Orion and Zanzibar). Overall, vaccination resulted in a significant difference in carriage of Salmonella between non-vaccinated and vaccinated commercial Cobb hens (P 0.05) could be determined for serogroup E. All vaccinated flocks produced a significant antibody response (P<0.001) to the S. Typhimurium vaccine strain, measured using a S. Typhimurium enzyme-linked immunosorbent assay (Guildhay), which peaked at 20 weeks of age, with 39% of the hens positive. Maternal antibodies were detected in 16% of the yolks from eggs produced by these flocks. There was a significant difference after challenge with Salmonella (P <0.05) among 1-day-old chicks from vaccinated versus non-vaccinated parents, when challenged using 10(4) CFU but not when challenged with 10(8) CFU. The success of this trial resulted in the incorporation of this vaccine into a Salmonella control system in commercial broiler breeder production.

Isolation of Salmonella spp. in cattle egrets (Bubulcus ibis) from Fernando de Noronha Archipelago, Brazil.

Science.gov (United States)

Silva, Marcio A; Fernandes, Érika F S T; Santana, Sandra C; Marvulo, Maria Fernanda V; Barros, Mércia R; Vilela, Sineide M O; Reis, Eliane M F; Mota, Rinaldo A; Silva, Jean C R

2018-03-03

The growth of the population of cattle egrets (Bubulcus ibis) in the archipelago of Fernando de Noronha constitutes a threat to public health and biological diversity because of their competition with and predation on native species and the possibility of transmission of pathogens to human beings, livestock and native wildlife. The aim here was to search for, isolate and identify serovars of Salmonella in clinically healthy local cattle egrets. Cloacal swabs were obtained from 456 clinically healthy cattle egrets of both sexes and a variety of ages. The swabs were divided into 51 pools. Six of these (11.7%) presented four serovars of Salmonella enterica subspecies enterica: Salmonella serovar Typhimurium; Salmonella serovar Newport; Salmonella serovar Duisburg; and Salmonella serovar Zega. One sample was identified as S. enterica subspecies enterica O16:y:-. Results in this study suggest that cattle egrets may be reservoirs of this agent on Fernando de Noronha and represent a risk to public health and biological diversity. Copyright © 2018. Published by Elsevier Editora Ltda.

Microbial radio-resistance of Salmonella Typhimurium in egg increases due to repetitive irradiation with electron beam

Energy Technology Data Exchange (ETDEWEB)

Tesfai, Adiam T.; Beamer, Sarah K.; Matak, Kristen E. [West Virginia University, Division of Animal and Nutritional Sciences, PO Box 6108, Morgantown, WV 26508 (United States); Jaczynski, Jacek, E-mail: Jacek.Jaczynski@mail.wvu.ed [West Virginia University, Division of Animal and Nutritional Sciences, PO Box 6108, Morgantown, WV 26508 (United States)

2011-04-15

Ionizing radiation improves food safety. However, foodborne pathogens develop increased resistance in response to sub-lethal stresses such as heat, pH, antibiotics, etc. Therefore, it is hypothesized that foodborne pathogens may develop increased radio-resistance to electron beam (e-beam) radiation. The objective was to determine if D{sub 10}-value for Salmonella Typhimurium in de-shelled raw egg (egg white and yolk mixed together) increases due to repetitive processing with e-beam at sub-lethal doses. Survivors were enumerated on non-selective (TSA) and selective (XLD) media. Survivors from the highest dose were isolated and used in subsequent e-beam cycle. This process was repeated four times for a total of five e-beam cycles. D{sub 10}-values for S. Typhimurium enumerated on TSA and XLD following each e-beam cycle were calculated as inverse reciprocal of the slope of survivor curves. D{sub 10}-values for the ATCC strain were 0.59{+-}0.031 and 0.46{+-}0.022 kGy on TSA and XLD, respectively. However, following the fifth e-beam cycle, the respective D{sub 10}-values increased (P<0.05) to 0.69{+-}0.026 and 0.61{+-}0.029 kGy, respectively. S. Typhimurium showed a trend (P>0.05) to develop radio-resistance faster on selective media, likely due to facilitated selection of radio-resistant cells within microbial population following each e-beam cycle. For all five e-beam cycles, S. Typhimurium had higher (P<0.05) D{sub 10}-values on non-selective media, indicating that sub-lethal injury followed by cellular repair and recovery are important for radio-resistance and inactivation of this microorganism. This study demonstrated that e-beam efficiently inactivates S. Typhimurium in raw egg; however, similar to other inactivation techniques and factors affecting microbial growth, S. Typhimurium develops increased radio-resistance if repetitively processed with e-beam at sub-lethal doses.

Serotypes and Antimicrobial Susceptibility of Salmonella spp. Isolated from Farm Animals in China

Directory of Open Access Journals (Sweden)

Yuan Zong Hui

2015-06-01

Full Text Available Salmonella spp. can indirectly infect humans via transfer from animals and animal-derived food products, and thereby cause potentially fatal diseases. Therefore, gaining an understanding of Salmonella infection in farm animals is increasingly important. The aim of this study was to identify the distribution of serotypes in Salmonella samples isolated from chickens (n = 837, pigs (n = 930, and dairy cows (n = 418 in central China (Henan, Hubei, and Hunan provinces in 2010–2011, and investigate the susceptibility of strains to antimicrobial agents. Salmonella isolates were identified by PCR amplification of the invA gene, serotypes were determined by using a slide agglutination test for O and H antigens, and susceptibility to 24 antimicrobials was tested using the agar dilution method. In total, 248 Salmonella strains were identified: 105, 105, and 38 from chickens, dairy cows, and pigs, respectively. Additionally, 209 strains were identified in unhealthy pigs from the Huazhong Agricultural University veterinary hospital. Among these 457 strains, the dominant serotypes were Typhimurium in serogroup B, IIIb in serogroup C, and Enteritidis in serogroup D. In antimicrobial susceptibility tests, 41.14% of Salmonella spp. were susceptible to all antimicrobial agents, 48.14% were resistant to at least one, and 34.72% were resistant to more than three classes. Strains were highly resistant to sulfamethoxazole-trimethoprim (39.61%, nalidixic acid (39.17%, doxycycline (28.22%, and tetracycline (27.58%. Resistance to cephalosporins and fluoroquinolones ranged from 5.25% to 7.44% and 19.04% to 24.51%, respectively. Among penicillin-resistant and cephalosporin-resistant strains, 25 isolates produced extended-spectrum β-lactamases (ESBLs. The multidrug-resistant and ESBL-producing Salmonella strains identified in healthy animals here will present a challenge for veterinary medicine and farm animal husbandry, and could also pose a threat to public health

[The occurrence of campylobacter fetus subsp. jejuni and Salmonella bacteria in some wild birds (author's transl)].

Science.gov (United States)

Rosef, O

1981-12-01

An investigation was carried out into the occurrence of Campylobacter fetus subsp. jejuni and Salmonella species in some wild birds. A total of 129 birds was examined, consisting of 71 pigeons, 54 seagulls, three crows and one raven. Campylobacter bacteria were isolated from 32 birds (24.8%), of which three were pigeons, 27 seagulls and two were crows. Of the 27 Campylobacter strains isolated from seagulls, four had the biochemical characteristics of the NARTC biotype described by Skirrow and Benjamin, seven were grouped as Campylobacter coli biotype and 16 as the biotype of Campylobacter jejuni. All the strains isolated from crows and pigeons had the biochemical characteristics of Campylobacter jejuni biotypes. Salmonella bacteria were isolated from the intestinal contents of two of the 54 seagulls (3.7%), and were identified serologically as Salmonella indiana and Salmonella typhimurium. One seagull was found to be a carrier of both Campylobacter fetus subsp. jejuni and Salmonella typhimurium. A correlation could not be demonstrated between the occurrence of Salmonella bacteria and Campylobacter fetus subsp. jejuni.

The prevalences of Salmonella Genomic Island 1 variants in human and animal Salmonella Typhimurium DT104 are distinguishable using a Bayesian approach.

Directory of Open Access Journals (Sweden)

Alison E Mather

Full Text Available Throughout the 1990 s, there was an epidemic of multidrug resistant Salmonella Typhimurium DT104 in both animals and humans in Scotland. The use of antimicrobials in agriculture is often cited as a major source of antimicrobial resistance in pathogenic bacteria of humans, suggesting that DT104 in animals and humans should demonstrate similar prevalences of resistance determinants. Until very recently, only the application of molecular methods would allow such a comparison and our understanding has been hindered by the fact that surveillance data are primarily phenotypic in nature. Here, using large scale surveillance datasets and a novel Bayesian approach, we infer and compare the prevalence of Salmonella Genomic Island 1 (SGI1, SGI1 variants, and resistance determinants independent of SGI1 in animal and human DT104 isolates from such phenotypic data. We demonstrate differences in the prevalences of SGI1, SGI1-B, SGI1-C, absence of SGI1, and tetracycline resistance determinants independent of SGI1 between these human and animal populations, a finding that challenges established tenets that DT104 in domestic animals and humans are from the same well-mixed microbial population.

Direct and indirect transmission of four Salmonella enterica serotypes in pigs

Directory of Open Access Journals (Sweden)

Österberg Julia

2010-05-01

Full Text Available Abstract Background Feed-borne spread of Salmonella spp. to pigs has been documented several times in recent years in Sweden. Experiences from the field suggest that feed-associated serotypes might be less transmittable and subsequently easier to eradicate from pig herds than other serotypes more commonly associated to pigs. Four Salmonella serotypes were selected for experimental studies in pigs in order to study transmissibility and compare possible differences between feed-assoociated (S Cubana and S Yoruba and pig-associated serotypes (S Derby and S Typhimurium. Methods Direct contact transmission was studied in four groups of pigs formed by six 10-week-old salmonella negative pigs commingled with two fatteners excreting one of the four salmonella serotypes. Indirect transmission was studied by putting six 10-week-old salmonella negative pigs in each of four salmonella contaminated rooms. Each room had previously housed a group of pigs, excreting one of the four selected serotypes. All pigs were monitored for two weeks with respect to the faecal excretion of salmonella and the presence of serum antibodies. At the end of the trial, eight samples from inner tissues and organs were collected from each pig at necropsy. Results In the four direct transmission groups, one pig shed Salmonella (Cubana at one occasion. At necropsy, S Typhimurium was isolated from one pig. In the indirect transmission groups, two pigs in the Yoruba room and one pig in each of the other rooms were excreting detectable levels of Salmonella once during the study period of two weeks. At necropsy, S Derby was isolated from one of six pigs in the Derby room and S Typhimurium was isolated from four of the six pigs in the Typhimurium room. No significant serological response could be detected in any of the 48 pigs. Conclusions These results show that all four selected serotypes were able to be transmitted in at least one of these field-like trials, but the transmission rate

LPS structure and PhoQ activity are important for Salmonella Typhimurium virulence in the Galleria mellonella infection model [corrected].

Directory of Open Access Journals (Sweden)

Jennifer K Bender

Full Text Available The larvae of the wax moth, Galleria mellonella, have been used experimentally to host a range of bacterial and fungal pathogens. In this study we evaluated the suitability of G. mellonella as an alternative animal model of Salmonella infection. Using a range of inoculum doses we established that the LD₅₀ of SalmonellaTyphimurium strain NCTC 12023 was 3.6 × 10³ bacteria per larva. Further, a set of isogenic mutant strains depleted of known virulence factors was tested to identify determinants essential for S. Typhimurium pathogenesis. Mutants depleted of one or both of the type III secretion systems encoded by Salmonella Pathogenicity Islands 1 and 2 showed no virulence defect. In contrast, we observed reduced pathogenic potential of a phoQ mutant indicating an important role for the PhoPQ two-component signal transduction system. Lipopolysaccharide (LPS structure was also shown to influence Salmonella virulence in G. mellonella. A waaL(rfaL mutant, which lacks the entire O-antigen (OAg, was virtually avirulent, while a wzz(ST/wzz(fepE double mutant expressing only a very short OAg was highly attenuated for virulence. Furthermore, shortly after infection both LPS mutant strains showed decreased replication when compared to the wild type in a flow cytometry-based competitive index assay. In this study we successfully established a G. mellonella model of S. Typhimurium infection. By identifying PhoQ and LPS OAg length as key determinants of virulence in the wax moth larvae we proved that there is an overlap between this and other animal model systems, thus confirming that the G. mellonella infection model is suitable for assessing aspects of Salmonella virulence function.

Plasma lipopolysaccharide level and enterocyte brush border enzymes in gnotobiotic piglets infected with Salmonella typhimurium

Czech Academy of Sciences Publication Activity Database

Trebichavský, Ilja; Kozáková, Hana; Šplíchal, Igor

2002-01-01

Roč. 47, - (2002), s. 289-294 ISSN 8750-7943 R&D Projects: GA ČR GA524/01/0917; GA AV ČR IAA5020101 Institutional research plan: CEZ:AV0Z5020903 Keywords : swine * gnotobiotic piglet * salmonella typhimurium Subject RIV: EE - Microbiology, Virology Impact factor: 0.107, year: 2002

A critical appraisal of the phene-plate biochemical fingerprinting system for epidemiological subtyping of Salmonella typhimurium

DEFF Research Database (Denmark)

On, S.L.W.; Baggesen, Dorte Lau

1996-01-01

The efficacy and reproducibility of the Phene-Plate (PhP) system (Biosys Inova, Stockholm, Sweden) for biochemical fingerprinting of Salmonella typhimurium was investigated. Duplicate and replicate assays on 40 epidemiologically related and unrelated strains were performed in two batches of PhP-48......P-types which are epidemiologically unjustified, (ii) tests currently recommended for PhP-typing S. typhimurium may be somewhat unstable and not satisfactory for fingerprinting purposes, (iii) caution must be exercised when comparing data from different batches of PhP-48 plates, and (iv) best results...

QUANTIFICATION OF Salmonella Typhimurium REDUCTION DURING COLD STORAGE OF RAW SHRIMPS IN THE PRESENCE OF SODIUM METABISULFITE [Kuantifikasi Reduksi Salmonella Typhimurium pada Udang Segar selama Penyimpanan Dingin dengan Penambahan Natrium Metabisulfit

Directory of Open Access Journals (Sweden)

Andiarto Yanuardi3

2012-12-01

Full Text Available Prediction of bacterial growth, survival or reduction in food matrices is needed for microbiological risk assessment. The survival of Salmonella Typhimurium on surfaces of raw shrimps at low temperature was studied, in the presence of sodium metabisulfite which is often used to prevent melanosis. The growth and/or reduction rates were quantified using DMFit software with Baranyi model and or linear model. The result showed that without sodium metabisulfite (control, when the initial level was high (105 CFU/ml, S. Typhimurium grew with a lag phase of 51.99±7.46 h and a growth rate of 0.01±0.002 log CFU.ml-1.h-1 on raw shrimps during storage at 8±2°C. When 1.5% (w/w sodium metabisulfite, a maximum level that often used to prevent melanosis, was added under the same condition, the number of S. Typhimurium was reduced for 5 log CFU/ml after 5 days, with a reduction rate of -0.03±0.001 log CFU.ml-1.h-1. This study indicated that Baranyi model can be used to predict the growth of S. Typhimurium on raw shrimp at low temperature, when sodium metabisulfite is absent. However, when sodium metabisulfite is present, at least 0.4% as found in this study, the reduction of S. Typhimurium can be predicted using a simple linear model.

Prevalence and Antimicrobial Resistance of Salmonella Isolates Recovered from Retail Pork in Major Village Markets in Tai'an Region, China.

Science.gov (United States)

Miao, Zengmin; Li, Song; Qin, Kun; Zhou, Yufa

2017-10-01

The current study was undertaken to evaluate Salmonella contamination in retail pork at major village markets of the Tai'an region, China. In total, 200 retail pork samples were collected from four village markets between June 2015 and February 2016, of which 69 samples (34.5%) were determined to be positive for Salmonella. Eleven serotypes were identified from the 69 Salmonella isolates, and Salmonella Derby was the most common (18 of 69, 26.1%), followed by Typhimurium (17 of 69, 24.6%) and Meleagridis (11 of 69, 15.9%). Antimicrobial susceptibility testing showed that antimicrobial resistance against tetracycline was the most prevalent (42 of 69, 60.9%), but antimicrobial resistance against both ceftriaxone and cefotaxime was 1.4% (1 of 69) and 2.9% (2 of 69), respectively. Multilocus sequence typing revealed that the 69 Salmonella isolates were divided into 11 sequence types (STs), among which ST40 (18 of 69, 26.1%) was the most common, followed by ST34 (15 of 69, 21.7%) and ST64 (13 of 69, 18.8%). Collectively, retail pork at village markets in the Tai'an region has a high Salmonella contamination rate, and these isolates exhibit broad-spectrum antimicrobial resistance. However, the absence of a dominant ST demonstrates that the Salmonella isolates from retail pork may be of diverse origins.

Characterization of Nontyphoidal Salmonella Isolates from Asymptomatic Migrant Food Handlers in Peninsular Malaysia.

Science.gov (United States)

Woh, Pei Yee; Thong, Kwai Lin; Behnke, Jerzy Marian; Lewis, John Watkin; Zain, Siti Nursheena Mohd

2017-08-01

Asymptomatic Salmonella carriers who work as food handlers pose food safety and public health risks, particularly during food preparation, and this has serious implications for the disease burden in society. Therefore, we conducted a study to determine the number of Salmonella carriers in a migrant cohort in several food establishments in three major cities in Peninsular Malaysia. Sociodemographic data and stool samples were collected and analyzed using standard methods of detection and isolation. Antimicrobial susceptibility tests of the positive samples were also performed. A total of 317 migrant food handlers, originating from South and Southeast Asian countries, were recruited voluntarily. Nine (2.8%) stool samples were confirmed to be Salmonella positive. PCR serotyping and pulsed-field gel electrophoresis identified four serotypes as Typhimurium (n = 3), Corvallis (n = 2), Hadar (n = 1), Agona (n = 1) and two unknown serovars. Antimicrobial susceptibility tests revealed that all nine isolates were susceptible to amoxicillin-clavulanic acid, cefotaxime, ceftazidime, ceftriaxone, and gentamycin. However, seven isolates were found to be multidrug resistant to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, sulfonamides, streptomycin, and tetracycline. This study highlights that carriers of nontyphoidal Salmonella exist among migrant food handlers, which poses a health risk to consumers through food contamination. Our results indicate a need for authorities to enhance food safety awareness in the migrant workers and to reevaluate current health screening methods to include preventive measure such as mandatory stool screening as part of the preemployment and routine health examinations.

Analysis of the protein profiles of the antibiotic-resistant Salmonella ...

African Journals Online (AJOL)

The emergent Salmonella typhimurium definitive phage type (DT) 104 is of particular global concern due to its frequent isolation and multiple antibiotic resistances. There is thus a need to know the kind of proteins expressed by S. typhimurium DT104 so as to provide a basis for developing an intervention. This study ...

Salmonella isolated from the feces of migrating cranes at the Izumi Plain (2002-2008): serotype, antibiotic sensitivity and PFGE type.

Science.gov (United States)

Kitadai, Noriyuki; Ninomiya, Naoko; Murase, Toshiyuki; Obi, Takeshi; Takase, Kozo

2010-07-01

From November 2002 to February 2008, 2,251 crane feces were collected at the Izumi Plain in Kagoshima Prefecture. Salmonella enterica was isolated from 359 feces (15.9%), of which 332 (92.5%) were Salmonella Typhimurium (ST), 9 were S. Hvittingfoss/II, 4 were S. Abaetetuba, 3 were S. Enteritidis, 2 were S. Konstanz, 1 was S. Pakistan and 8 were untyped isolates, respectively. Against 12 antimicrobial agents, no resistant strains were found in 154 isolates examined, but one was found to be resistant to ampicillin. By pulsed-field gel electrophoresis (PFGE), all but one of the 68 ST isolates tested showed indistinguishable banding patterns; one had a different pattern. The results suggest that ST strains from the same origin would spread in crane flocks during their stay at Izumi Plain every winter.

The serologic response to Salmonella enteritidis and Salmonella typhimurium in experimentally infected chickens, followed by an indirect lipopolysaccharide enzyme-linked immunosorbent assay and bacteriologic examinations through a one-year period

DEFF Research Database (Denmark)

Skov, M.N.; Feld, Niels Christian; Carstensen, B.

2002-01-01

Three groups of 100 individually marked salmonella-free chickens were followed for a period of 53 wk. The chickens were infected as day olds by crop instillation of 101 colony-forming units: one group with Salmonella enteritidis and a second group with Salmonella typhimurium. A third group was kept...... in surveillance programs, in particular to detect flocks in early stages of infection before a measurable serologic response has been raised....

Modified intracellular-associated phenotypes in a recombinant Salmonella Typhi expressing S. Typhimurium SPI-3 sequences.

Directory of Open Access Journals (Sweden)

Patricio Retamal

Full Text Available A bioinformatics comparison of Salmonella Pathogenicity Island 3 sequences from S. Typhi and S. Typhimurium serovars showed that ten genes are highly conserved. However three of them are pseudogenes in S. Typhi. Our aim was to understand what functions are lost in S. Typhi due to pseudogenes by constructing a S. Typhi genetic hybrid carrying the SPI-3 region of S. Typhimurium instead of its own SPI-3. We observed that under stressful conditions the hybrid strain showed a clear impairment in resistance to hydrogen peroxide and decreased survival within U937 culture monocytes. We hypothesized that the marT-fidL operon, encoded in SPI-3, was responsible for the new phenotypes because marT is a pseudogen in S. Typhi and has a demonstrated role as a transcriptional regulator in S. Typhimurium. Therefore we cloned and transferred the S. Typhimurium marT-fidL operon into S. Typhi and confirmed that invasion of monocytes was dramatically decreased. Finally, our findings suggest that the genomic and functional differences between SPI-3 sequences have implications in the host specificity of Typhi and Typhimurium serovars.

Isolation and characterization of polyvalent bacteriophages infecting multi drug resistant Salmonella serovars isolated from broilers in Egypt.

Science.gov (United States)

Mahmoud, Mayada; Askora, Ahmed; Barakat, Ahmed Barakat; Rabie, Omar El-Farouk; Hassan, Sayed Emam

2018-02-02

In this study, we isolated and characterized three phages named as Salmacey1, Salmacey2 and Salmacey3, infecting multi drug resistant Salmonella serovars isolated from broilers in Egypt. The most prevalent Salmonella serovars were S. typhimurium, S. enteritidis, and S. kentucky. All these Salmonella serovars were found to be resistant to more than two of the ten antimicrobial agents tested. Only S. kentucky was found to be resistant to seven antimicrobial agents. Examination of these phage particles by transmission electron microscopy (TEM), demonstrated that two phages (Salmacey1, Salmacey2) were found to belong to family Siphoviridae, and Salmacey3 was assigned to the family Myoviridae. The results of host range assay revealed that these bacteriophages were polyvalent and thus capable of infecting four strains of Salmonella serovars and Citrobacter freundii. Moreover, the two phages (Salmacey1, Salmacey2) had a lytic effect on Enterobacter cloacae and Salmacey3 was able to infect E. coli. All phages could not infect S. para Typhi, Staphylococus aureus and Bacillus cereus. One-step growth curves of bacteriophages revealed that siphovirus phages (Salmacey1, Salmacey2) have burst size (80 and 90pfu per infected cell with latent period 35min and 40min respectively), and for the myovirus Salmacey3 had a burst size 110pfu per infected cell with latent period 60min. Molecular analyses indicated that these phages contained double-stranded DNA genomes. The lytic activity of the phages against the most multidrug resistant serovars S. kentucky as host strain was evaluated. The result showed that these bacteriophages were able to completely stop the growth of S. kentucky in vitro. These results suggest that phages have a high potential for phage application to control Salmonella serovars isolated from broilers in Egypt. Copyright © 2017. Published by Elsevier B.V.

Salmonella enterica Typhimurium fljBA operon stability: implications regarding the origin of Salmonella enterica I 4,[5],12:i:.

Science.gov (United States)

Tomiyama, M P O; Werle, C H; Milanez, G P; Nóbrega, D B; Pereira, J P; Calarga, A P; Flores, F; Brocchi, M

2015-12-29

Salmonella enterica subsp enterica serovar 4,5,12:i:- has been responsible for many recent Salmonella outbreaks worldwide. Several studies indicate that this serovar originated from S. enterica subsp enterica serovar Typhimurium, by the loss of the flagellar phase II gene (fljB) and adjacent sequences. However, at least two different clones of S. enterica 4,5,12:i:- exist that differs in the molecular events responsible for fljB deletion. The aim of this study was to test the stability of the fljBA operon responsible for the flagellar phase variation under different growth conditions in order to verify if its deletion is a frequent event that could explain the origin and dissemination of this serovar. In fact, coding sequences for transposons are present near this operon and in some strains, such as S. enterica Typhimurium LT2, the Fels-2 prophage gene is inserted near this operon. The presence of mobile DNA could confer instability to this region. In order to examine this, the cat (chloramphenicol acetyltransferase) gene was inserted adjacent to the fljBA operon so that deletions involving this genomic region could be identified. After growing S. enterica chloramphenicol-resistant strains under different conditions, more than 104 colonies were tested for the loss of chloramphenicol resistance. However, none of the colonies were sensitive to chloramphenicol. These data suggest that the origin of S. enterica serovar 4,5,12:i:- from Typhimurium by fljBA deletion is not a frequent event. The origin and dissemination of 4,5,12:i:- raise several questions about the role of flagellar phase variation in virulence.

Antibiotic resistance determinants and genetic analysis of Salmonella enterica isolated from food in Morocco.

Science.gov (United States)

Murgia, Manuela; Bouchrif, Brahim; Timinouni, Mohammed; Al-Qahtani, Ahmed; Al-Ahdal, Mohammed N; Cappuccinelli, Pietro; Rubino, Salvatore; Paglietti, Bianca

2015-12-23

Antimicrobial-resistant non-typhoidal Salmonella (NTS) are an important cause of infection in Africa, but there is a lack of information on their molecular mechanisms of resistance and epidemiology. This study contributes to fill this gap through the characterization by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), plasmid profiling and analysis of antibiotic-resistance determinants of 94 Salmonella enterica strains isolated from food in Morocco. PFGE revealed considerable heterogeneity among the strains, showing 32 pulsotypes. MLST of strains representative of the different serovars evidenced 13 sequence types (STs), three of which were newly identified (ST1694, ST1768 and ST1818) and nine not previously reported in Morocco. Thirty-four strains harbored from one to four plasmids, of IncI1 group in S. Mbandaka, IncFIIA in S. Typhimurium, IncL/M in S. Hadar and S. Blockley. For the first time in Morocco an intact Salmonella Genomic Island 1 (SGI1) carrying the resistance genes aadA2, floR, tetG, blaPSE-1 and sul1 was detected in S. Typhimurium DT104. In serovar Hadar resistance to ampicillin, tetracycline and streptomycin was associated to blaTEM-1, tetA and strA genes respectively, whereas one mutation in gyrA (Asp87Asn) and one in parC (Thr54Ser) genes conferred resistance to nalidixic acid. These findings improve the information on foodborne Salmonella in Morocco, evidencing the presence of MDR strains potentially dangerous to humans, and provide useful data for future studies. Copyright © 2015 Elsevier B.V. All rights reserved.

A functional cra gene is required for Salmonella enterica serovar typhimurium virulence in BALB/c mice

DEFF Research Database (Denmark)

Allen, J. H.; Utley, M.; Van den Bosch, H.

2000-01-01

A minitransposon mutant of Salmonella enterica serovar Typhimurium SR-11, SR-11 Fad(-), is unable to utilize gluconeogenic substrates as carbon sources and is avirulent and immunogenic when administered perorally to BALB/c mice (M. J. Utley et al., FEMS Microbiol. Lett., 163:129-134, 1998). Here,...

X-Ray crystal structure of GarR—tartronate semialdehyde reductase from Salmonella typhimurium

OpenAIRE

Osipiuk, J.; Zhou, M.; Moy, S.; Collart, F.; Joachimiak, A.

2009-01-01

Tartronate semialdehyde reductases (TSRs), also known as 2-hydroxy-3-oxopropionate reductases, catalyze the reduction of tartronate semialdehyde using NAD as cofactor in the final stage of D-glycerate biosynthesis. These enzymes belong to family of structurally and mechanically related β-hydroxyacid dehydrogenases which differ in substrate specificity and catalyze reactions in specific metabolic pathways. Here, we present the crystal structure of GarR a TSR from Salmonella typhimurium determi...

Comparative proteomic analysis of Salmonella enterica serovar Typhimurium ppGpp-deficient mutant to identify a novel virulence protein required for intracellular survival in macrophages

Directory of Open Access Journals (Sweden)

Kumagai Yoshinori

2010-12-01

Full Text Available Abstract Background The global ppGpp-mediated stringent response in pathogenic bacteria plays an important role in the pathogenesis of bacterial infections. In Salmonella enterica serovar Typhimurium (S. Typhimurium, several genes, including virulence genes, are regulated by ppGpp when bacteria are under the stringent response. To understand the control of virulence genes by ppGpp in S. Typhimurium, agarose 2-dimensional electrophoresis (2-DE combined with mass spectrometry was used and a comprehensive 2-DE reference map of amino acid-starved S. Typhimurium strain SH100, a derivative of ATCC 14028, was established. Results Of the 366 examined spots, 269 proteins were successfully identified. The comparative analysis of the wild-type and ppGpp0 mutant strains revealed 55 proteins, the expression patterns of which were affected by ppGpp. Using a mouse infection model, we further identified a novel virulence-associated factor, STM3169, from the ppGpp-regulated and Salmonella-specific proteins. In addition, Salmonella strains carrying mutations in the gene encoding STM3169 showed growth defects and impaired growth within macrophage-like RAW264.7 cells. Furthermore, we found that expression of stm3169 was controlled by ppGpp and SsrB, a response regulator of the two-component system located on Salmonella pathogenicity island 2. Conclusions A proteomic approach using a 2-DE reference map can prove a powerful tool for analyzing virulence factors and the regulatory network involved in Salmonella pathogenesis. Our results also provide evidence of a global response mediated by ppGpp in S. enterica.

Epidemiological characteristics of Salmonella Typhimurium isolated from animals and feed in Poland

DEFF Research Database (Denmark)

Wasyl, D.; Sandvang, D.; Skov, M. N.

2006-01-01

of XbaI and BlnI digest showed high genomic similarity between the strains and confirmed clonal spread of S. Typhimurium infections. Plasmid profiling allowed further differentiation of the strains. We have, therefore, confirmed the appearance of S. Typhimurium DT104 showing genome integrated integron......-mediated antimicrobial resistance in Poland. These findings are significant for public and animal health risks and document the dissemination of DT104 epidemic strains into new geographical regions....

Cloning and properties of the Salmonella typhimurium tricarboxylate transport operon in Escherichia coli

International Nuclear Information System (INIS)

Widenhorn, K.A.; Boos, W.; Somers, J.M.; Kay, W.W.

1988-01-01

The tricarboxylate transport operon (tctI) was cloned in Escherichia coli as a 12-kilobase (kb) fragment from an EcoRI library of the Salmonella typhimurium chromosome in λgtWES. It was further subcloned as a 12-kb fragment into pACYC184 and as an 8-kb fragment into pBR322. By insertional mutagenesis mediated by λTn5, restriction mapping, and phenotypic testing, the tctI operon was localized to a 4.5-kb region. The tctC gene which encodes a periplasmic binding protein (C-protein) was located near the center of the insert. E. coli/tctI clones on either multicopy or single-copy vectors grew on the same tricarboxylates as S. typhimurium, although unusually long growth lags were observed. E. coli/tctI clones exhibited similar [ 14 C] fluorocitrate transport kinetics to those of S. typhimurium, whereas E. coli alone was virtually impermeable to [ 14 C] fluorocitrate. The periplasmic C proteins (C1 and C2 isoelectric forms) were produced in prodigious quantities from the cloned strains. Motile E. coli/tctI clones were not chemotactic toward citrate, whereas tctI deletion mutants of S. typhimurium were. Taken together, these observations indicate that tctI is not an operon involved in chemotaxis

Genetic characterisation of multidrug-resistant Salmonella enterica serotypes isolated from poultry in Cairo, Egypt

Directory of Open Access Journals (Sweden)

Mohammed Abdel-Maksoud

2015-05-01

Full Text Available Background: Food-borne diseases pose serious health problems, affecting public health and economic development worldwide. Methods: Salmonella was isolated from samples of chicken parts, skin samples of whole chicken carcasses, raw egg yolks, eggshells and chicken faeces. Resulting isolates were characterised by serogrouping, serotyping, antimicrobial susceptibility testing and detection of extended-spectrum β-lactamase (ESBL production. Antibiotic resistance genes and integrons were identified by polymerase chain reaction (PCR. Results: The detection rates of Salmonella were 60%, 64% and 62% in chicken parts, skin, and faeces, respectively, whereas the egg yolks and eggshells were uniformly negative. Salmonella Kentucky and S. Enteritidis serotypes comprised 43.6% and 2.6% of the isolates, respectively, whilst S. Typhimurium was absent. Variable resistance rates were observed against 16 antibiotics; 97% were resistant to sulfamethoxazole, 96% to nalidixic acid and tetracycline and 76% to ampicillin. Multidrug resistance was detected in 82% (64/78 of the isolates and ESBL production was detected in 8% (6/78. The β-lactamase blaTEM-1 gene was detected in 57.6% and blaSHV-1 in 6.8% of the isolates, whilst the blaOXA gene was absent. The sul1gene was detected in 97.3% and the sul2 gene in 5.3% of the isolates. Sixty-four of the 78 isolates (82% were positive for the integrase gene (int I from class 1 integrons, whilst int II was absent. Conclusion: This study reveals the presence of an alarming number of multidrug-resistant Salmonella isolates in the local poultry markets in Cairo. The high levels of drug resistance suggest an emerging problem that could impact negatively on efforts to prevent and treat poultry and poultry-transmitted human diseases in Egypt.

Phenotypic and genotypic antimicrobial resistance and virulence genes of Salmonella enterica isolated from pet dogs and cats

Science.gov (United States)

Srisanga, Songsak; Angkititrakul, Sunpetch; Sringam, Patcharee; Le Ho, Phuong T.; Vo, An T. T.

2017-01-01

Salmonella enterica isolates (n = 122), including 32 serotypes from 113 dogs and 9 cats, were obtained from household dogs (n = 250) and cats (n = 50) during 2012–2015. The isolates were characterized by serotyping, antimicrobial resistance phenotyping and genotyping, and virulence gene screening. Serovars Weltevreden (15.6%) and Typhimurium (13.9%) were the most common. The majority (43%) of the isolates were multidrug resistant. The dog isolates (12.3%) harbored class 1 integrons, of which the dfrA12-aadA2 cassette was most frequent (66.7%). The only class integron in serovar Albany was located on a conjugative plasmid. Two ESBL-producing isolates (i.e., a serovar Krefeld and a serovar Enteritridis) carried blaTEM and blaCTX-M, and the blaTEM gene in both was horizontally transferred. Of the plasmid-mediated quinolone resistance genes tested, only qnrS (4.9%) was detected. Most Salmonella isolates harbored invA (100%), prgH (91.8%), and sipB (91%). Positive associations between resistance and virulence genes were observed for blaPSE-1/orgA, cmlA/spaN, tolC, and sul1/tolC (p resistance and virulence genes and that antimicrobial use in companion animals may select for the examined Salmonella virulence factors. PMID:27586467

Effects of commercial marinade seasoning and a natural blend of cultured sugar and vinegar on Campylobacter jejuni and Salmonella Typhimurium and the texture of chicken breasts.

Science.gov (United States)

Park, Na Yoon; Hong, Soo Hyeon; Yoon, Ki Sun

2014-03-01

Marination using various ingredients has been widely used to improve microbial safety and quality of chicken products at retail markets. The objective of this study was to investigate the effects of commercial marinade seasoning and cultured sugar/vinegar blend on Campylobacter jejuni and Salmonella Typhimurium populations during refrigerated storage. In addition, their effects on the texture of precooked chicken breasts during frozen and refrigerated storage was investigated. Chicken breasts inoculated with 4.5 to 5.0 log cfu/g of C. jejuni and Salmonella Typhimurium were treated with 3% cultured sugar/vinegar blend with and without 0.6% polish rub seasoning containing 32% herb content. Breasts were then vacuum-packaged and stored at 4 and 10°C. Survival and growth curves were fitted to the Baranyi equation to determine survival and growth kinetics of C. jejuni and Salmonella Typhimurium. In addition, the vacuum-packaged precooked chicken breasts with different marination treatments were subjected to 3 freeze-thaw cycles and shear force was measured. At 4°C, the populations of C. jejuni and Salmonella Typhimurium decreased, regardless of treatment group during storage. The greatest survival for C. jejuni was observed in untreated chicken breasts. At 10°C, the growth of Salmonella Typhimurium was completely prevented in precooked chicken breasts treated with 3% cultured sugar/vinegar blend, regardless of the presence of 0.6% seasoning. The 3% cultured sugar/vinegar blend also improved the tenderness of frozen chicken breasts and refrigerated, ready-to-eat chicken breast. Therefore, a natural blend of cultured sugar and vinegar can be used as antimicrobial and texture-modifying agents for poultry meat and poultry products.

Identification of Metabolic Pathways Essential for Fitness of Salmonella Typhimurium In Vivo

DEFF Research Database (Denmark)

Jelsbak, Lotte; Hartman, Hassan; Schroll, Casper

2014-01-01

Bacterial infections remain a threat to human and animal health worldwide, and there is an urgent need to find novel targets for intervention. In the current study we used a computer model of the metabolic network of Salmonella enterica serovar Typhimurium and identified pairs of reactions (cut s...

Transcriptomic analysis of swarm motility phenotype of Salmonella enterica serovar Typhimurium mutant defective in periplasmic glucan synthesis

Science.gov (United States)

Movement of food-borne pathogens on moist surfaces enables them to migrate towards more favorable niches and facilitate their survival for extended periods of time. Salmonella enterica serovar Typhimurium mutants defective in OPG synthesis are unable to exhibit motility on moist surfaces (swarming) ...

A Portable Impedance Immunosensing System for Rapid Detection of Salmonella Typhimurium.

Science.gov (United States)

Wen, Tao; Wang, Ronghui; Sotero, America; Li, Yanbin

2017-08-28

Salmonella Typhimurium is one of the most dangerous foodborne pathogens and poses a significant threat to human health. The objective of this study was to develop a portable impedance immunosensing system for rapid and sensitive detection of S . Typhimurium in poultry. The developed portable impedance immunosensing system consisted of a gold interdigitated array microelectrode (IDAM), a signal acquisitive interface and a laptop computer with LabVIEW software. The IDAM was first functionalized with 16-Mercaptohexadecanoic acid, and streptavidin was immobilized onto the electrode surface through covalent bonding. Then, biotin-labelled S . Typhimurium -antibody was immobilized onto the IDAM surface. Samples were dropped on the surface of the IDAM and the S . Typhimurium cells in the samples were captured by the antibody on the IDAM. This resulted in impedance changes that were measured and displayed with the LabVIEW software. An equivalent circuit of the immunosensor demonstrated that the largest change in impedance was due to the electron-transfer resistance. The equivalent circuit showed an increase of 35% for the electron-transfer resistance value compared to the negative control. The calibration result indicated that the portable impedance immunosensing system could be used to measure the standard impedance elements, and it had a maximum error of measurement of approximately 13%. For pure culture detection, the system had a linear relationship between the impedance change and the logarithmic value of S . Typhimurium cells ranging from 76 to 7.6 × 10⁶ CFU (colony-forming unit) (50 μL) -1 . The immunosensor also had a correlation coefficient of 0.98, and a high specificity for detection of S . Typhimurium cells with a limit of detection (LOD) of 10² CFU (50 μL) -1 . The detection time from the moment a sample was introduced to the display of the results was 1 h. To conclude, the portable impedance immunosensing system for detection of S . Typhimurium achieved

A Portable Impedance Immunosensing System for Rapid Detection of Salmonella Typhimurium

Directory of Open Access Journals (Sweden)

Tao Wen

2017-08-01

Full Text Available Salmonella Typhimurium is one of the most dangerous foodborne pathogens and poses a significant threat to human health. The objective of this study was to develop a portable impedance immunosensing system for rapid and sensitive detection of S. Typhimurium in poultry. The developed portable impedance immunosensing system consisted of a gold interdigitated array microelectrode (IDAM, a signal acquisitive interface and a laptop computer with LabVIEW software. The IDAM was first functionalized with 16-Mercaptohexadecanoic acid, and streptavidin was immobilized onto the electrode surface through covalent bonding. Then, biotin-labelled S. Typhimurium-antibody was immobilized onto the IDAM surface. Samples were dropped on the surface of the IDAM and the S. Typhimurium cells in the samples were captured by the antibody on the IDAM. This resulted in impedance changes that were measured and displayed with the LabVIEW software. An equivalent circuit of the immunosensor demonstrated that the largest change in impedance was due to the electron-transfer resistance. The equivalent circuit showed an increase of 35% for the electron-transfer resistance value compared to the negative control. The calibration result indicated that the portable impedance immunosensing system could be used to measure the standard impedance elements, and it had a maximum error of measurement of approximately 13%. For pure culture detection, the system had a linear relationship between the impedance change and the logarithmic value of S. Typhimurium cells ranging from 76 to 7.6 × 106 CFU (colony-forming unit (50 μL−1. The immunosensor also had a correlation coefficient of 0.98, and a high specificity for detection of S. Typhimurium cells with a limit of detection (LOD of 102 CFU (50 μL−1. The detection time from the moment a sample was introduced to the display of the results was 1 h. To conclude, the portable impedance immunosensing system for detection of S. Typhimurium

Distribution and Antimicrobial Susceptibility of Foodborne Salmonella Serovars in Eight Provinces in China from 2007 to 2012 (Except 2009).

Science.gov (United States)

Wang, Yin; Cao, Chenyang; Alali, Walid Q; Cui, Shenghui; Li, Fengqin; Zhu, Jianghui; Wang, Xin; Meng, Jianghong; Yang, Baowei

2017-07-01

One thousand four hundred ninety-one Salmonella isolates recovered from retail foods including chicken, beef, fish, pork, dumplings, and cold dishes in China in 2007, 2008, 2010, 2011, and 2012 were analyzed for distribution of serotype and antimicrobial susceptibility. A total of 129 Salmonella serotypes were detected among 1491 isolates. Salmonella Enteritidis (21.5%), Typhimurium (11.0%), Indiana (10.8%), Thompson (5.4%), Derby (5.1%), Agona (3.8%), and Shubra (3.0%) were the seven most important serotypes in 1491 isolates. For antibiotic susceptibility, except 16 (1.1%) isolates were susceptible to all tested antibiotics, 131 (8.8%) resisted 1-2 and 1344 (90.1%) resisted three or more antibiotics. One thousand forty-six (70.2%) of 1491 Salmonella isolates were identified as multidrug-resistant (MDR) isolates, which could resist three or more categories of antibiotics. Resistance to sulfisoxazole (78.1%) was most common among the tested Salmonella, followed by tetracycline (70.6%), trimethoprim/sulfamethoxazole (68.0%), and nalidixic acid (63.4%). Resistances to amikacin (20.0%), levofloxacin (18.7%), gatifloxacin (17.9%), ceftriaxone (17.7%), and cefoxitin (13.2%) were less frequently detected. Resistance to fluoroquinolones was most common among Salmonella Shubra and Indiana isolates, while resistance to cephalosporins was frequently detected among Salmonella Thompson isolates. The results highlighted the diversity of Salmonella serotypes and the high prevalence of Salmonella MDR isolates in China. Compared with Salmonella Enteritidis and Typhimurium isolates, the higher fluoroquinolones and cephalosporins resistance rates of some individual serotypes (Salmonella Shubra, Indiana, and Thompson) also provided more information for further study related to fluoroquinolones or cephalosporin-resistant Salmonella.

Evidence of metabolic switching and implications for food safety from the phenome(s) of Salmonella enterica serovar Typhimurium DT104 cultured at selected points across the pork production food chain.

Science.gov (United States)

Martins, Marta; McCusker, Matthew P; McCabe, Evonne M; O'Leary, Denis; Duffy, Geraldine; Fanning, Séamus

2013-09-01

Salmonella enterica serovar Typhimurium DT104 is a recognized food-borne pathogen that displays a multidrug-resistant phenotype and that is associated with systemic infections. At one extreme of the food chain, this bacterium can infect humans, limiting the treatment options available and thereby contributing to increased morbidity and mortality. Although the antibiotic resistance profile is well defined, little is known about other phenotypes that may be expressed by this pathogen at key points across the pork production food chain. In this study, 172 Salmonella enterica serovar Typhimurium DT104/DT104b isolated from an extensive "farm-to-fork" surveillance study, focusing on the pork food chain, were characterized in detail. Isolates were cultured from environmental, processing, retail, and clinical sources, and the study focused on phenotypes that may have contributed to persistence/survival in these different niches. Molecular subtypes, along with antibiotic resistance profiles, tolerance to biocides, motility, and biofilm formation, were determined. As a basis for human infection, acid survival and the ability to utilize a range of energy sources and to adhere to and/or invade Caco-2 cells were also studied. Comparative alterations to biocide tolerance were observed in isolates from retail. l-Tartaric acid and d-mannose-1-phosphate induced the formation of biofilms in a preselected subset of strains, independent of their origin. All clinical isolates were motile and demonstrated an enhanced ability to survive in acidic conditions. Our data report on a diverse phenotype, expressed by S. Typhimurium isolates cultured from the pork production food chain. Extending our understanding of the means by which this pathogen adapts to environmental niches along the "farm-to-fork" continuum will facilitate the protection of vulnerable consumers through targeted improvements in food safety measures.

Safety and tolerability of a live oral Salmonella typhimurium vaccine candidate in SIV-infected nonhuman primates.

Science.gov (United States)

Ault, Alida; Tennant, Sharon M; Gorres, J Patrick; Eckhaus, Michael; Sandler, Netanya G; Roque, Annelys; Livio, Sofie; Bao, Saran; Foulds, Kathryn E; Kao, Shing-Fen; Roederer, Mario; Schmidlein, Patrick; Boyd, Mary Adetinuke; Pasetti, Marcela F; Douek, Daniel C; Estes, Jacob D; Nabel, Gary J; Levine, Myron M; Rao, Srinivas S

2013-12-02

Nontyphoidal Salmonella (NTS) serovars are a common cause of acute food-borne gastroenteritis worldwide and can cause invasive systemic disease in young infants, the elderly, and immunocompromised hosts, accompanied by high case fatality. Vaccination against invasive NTS disease is warranted where the disease incidence and mortality are high and multidrug resistance is prevalent, as in sub-Saharan Africa. Live-attenuated vaccines that mimic natural infection constitute one strategy to elicit protection. However, they must particularly be shown to be adequately attenuated for consideration of immunocompromised subjects. Accordingly, we examined the safety and tolerability of an oral live attenuated Salmonella typhimurium vaccine candidate, CVD 1921, in an established chronic simian immunodeficiency virus (SIV)-infected rhesus macaque model. We evaluated clinical parameters, histopathology, and measured differences in mucosal permeability to wild-type and vaccine strains. Compared to the wild-type S. typhimurium strain I77 in both SIV-infected and SIV-uninfected nonhuman primate hosts, this live-attenuated vaccine shows reduced shedding and systemic spread, exhibits limited pathological disease manifestations in the digestive tract, and induces low levels of cellular infiltration in tissues. Furthermore, wild-type S. typhimurium induces increased intestinal epithelial damage and permeability, with infiltration of neutrophils and macrophages in both SIV-infected and SIV-uninfected nonhuman primates compared to the vaccine strain. Based on shedding, systemic spread, and histopathology, the live-attenuated S. typhimurium strain CVD 1921 appears to be safe and well-tolerated in the nonhuman primate model, including chronically SIV-infected rhesus macaques. Copyright © 2013. Published by Elsevier Ltd.

Metabolic profiling of meat: assessment of pork hygiene and contamination with Salmonella typhimurium.

Science.gov (United States)

Xu, Yun; Cheung, William; Winder, Catherine L; Dunn, Warwick B; Goodacre, Royston

2011-02-07

Spoilage in meat is the result of the action of microorganisms and results in changes of meat and microbial metabolism. This process may include pathogenic food poisoning bacteria such as Salmonella typhimurium, and it is important that these are differentiated from the natural spoilage process caused by non-pathogenic microorganisms. In this study we investigated the application of metabolic profiling using gas chromatography-mass spectrometry, to assess the microbial contamination of pork. Metabolite profiles were generated from microorganisms, originating from the natural spoilage process and from the artificial contamination with S. typhimurium. In an initial experiment, we investigated changes in the metabolic profiles over a 72 hour time course at 25 °C and established time points indicative of the spoilage process. A further experiment was performed to provide in-depth analysis of the metabolites characteristic of contamination by S. typhimurium. We applied a three-way PARAllel FACtor analysis 2 (PARAFAC2) multivariate algorithm to model the metabolic profiles. In addition, two univariate statistical tests, two-sample Wilcoxon signed rank test and Friedman test, were employed to identify metabolites which showed significant difference between natural spoiled and S. typhimurium contaminated samples. Consistent results from the two independent experiments were obtained showing the discrimination of the metabolic profiles of the natural spoiled pork chops and those contaminated with S. typhimurium. The analysis identified 17 metabolites of significant interest (including various types of amino acid and fatty acid) in the discrimination of pork contaminated with the pathogenic microorganism.

Eradication of Salmonella Typhimurium infection in a murine model of typhoid fever with the combination of probiotic Lactobacillus fermentum ME-3 and ofloxacin

Directory of Open Access Journals (Sweden)

Karki Tõnis

2008-08-01

Full Text Available Abstract Background The aim of the study was to detect whether in experimental Salmonella enterica Typhimurium infection the probiotic Lactobacillus fermentum ME-3 in combination with fluoroquinolone therapy would eradicate S. Typhimurium, prevent the development of liver and spleen granulomas and improve the indices of oxidative stress in the ileum mucosa. The selected bacteriological, histological and biochemical methods were applied. Results Combined treatment with L. fermentum ME-3 and ofloxacin eradicated Salmonella Typhimurium from blood, ileum and liver, decreased the number of animals with liver and spleen granulomas and reduced the value of lipid peroxides in the ileum mucosa. Higher total counts of intestinal lactobacilli in all experimental groups were associated with the absence of liver granulomas. Conclusion The antimicrobial and antioxidative probiotic L. fermentum ME-3 combined with ofloxacin enhances the eradication of experimental S. Typhimurium infection. These observations on probiotic and antimicrobial co-action may serve as basis to develop new strategies for treatment of invasive bacterial infections of the gut.

Tackling the issue of environmental survival of live Salmonella Typhimurium vaccines: deletion of the lon gene.

Science.gov (United States)

Leyman, Bregje; Boyen, Filip; Van Parys, Alexander; Verbrugghe, Elin; Haesebrouck, Freddy; Pasmans, Frank

2012-12-01

Vaccination is an important measure to control Salmonella contamination in the meat production chain. A previous study showed that both the ΔrfaJ and ΔrfaL strains are suitable markers and allow serological differentiation of infected and vaccinated animals. The aim of this study was to verify whether deletion of the lon gene in a Salmonella Typhimurium ΔrfaJ marker strain resulted in decreased environmental survival. Our results indicate that deletion of the lon gene in the ΔrfaJ strain did not affect invasiveness in IPEC-J2 cells and resulted in an increased susceptibility to UV, disinfectants (such as hydrogen peroxide and tosylchloramide sodium) and citric acid. Immunization of pigs with inactivated ΔrfaJ or ΔlonΔrfaJ vaccines allowed differentiation of infected and vaccinated pigs. Furthermore, deletion of the lon gene did not reduce the protection conferred by live wild type or ΔrfaJ vaccines against subsequent challenge with a virulent Salmonella Typhimurium strain in BALB/c mice. Based on our results in mice, we conclude that deletion of lon in ΔrfaJ contributes to environmental safety of the ΔrfaJ DIVA strain. Copyright © 2012 Elsevier Ltd. All rights reserved.

Laboratory monitoring of bacterial gastroenteric pathogens Salmonella and Shigella in Shanghai, China 2006-2012.

Science.gov (United States)

Zhang, J; Wang, F; Jin, H; Hu, J; Yuan, Z; Shi, W; Yang, X; Meng, J; Xu, X

2015-02-01

In 2006 we initiated an enhanced laboratory-based surveillance of Salmonella and Shigella infections in Shanghai, China. A total of 4483 Salmonella and 2226 Shigella isolates were recovered from stool specimens by 2012. In 80 identified Salmonella serovars, Enteritidis (34·5%) and Typhimurium (26·2%) were the most common. Shigella (S.) sonnei accounted for 63·9% of human Shigella infections over the same time period, and replaced S. flexneri to become the primary cause of shigellosis since 2010. Overall, a high level of antimicrobial resistance was observed in Salmonella and Shigella, particularly to nalidixic acid, ampicillin, and tetracycline. Ciprofloxacin resistance was common in Salmonella Typhimurium (21·0%) and S. flexneri (37·6%). The cephalosporin resistance in both pathogens also increased over the years, ranging from 3·4% to 7·0% in Salmonella, and from 10·4% to 28·6% in Shigella. Resistance to multiple antimicrobials was also identified in a large number of the isolates. This study provides insight into the distribution of Salmonella and Shigella in diarrhoeal diseases.

Buffer capacity of food components influences the acid tolerance response in Salmonella Typhimurium during simulated gastric passage

DEFF Research Database (Denmark)

Henriksen, Sidsel; Buschhardt, Tasja; Hansen, Tina Beck

2014-01-01

tubes, enabling simultaneous testing of biological triplicates under varying conditions. Surprisingly, we found that less buffered media provided higher protection of Salmonella, compared to media with high buffer capacity. By investigating the relative gene expression of rpoS and ompR encoding for two...... Heart Infusion Broth having a higher buffer capacity. We suggest this to be associated with a varying ability of Salmonella Typhimurium to mount a stationary phase acid tolerance response (ATR) depending on the buffer capacity of the food vehicle....

The architecture and ppGpp-dependent expression of the primary transcriptome of Salmonella Typhimurium during invasion gene expression

Directory of Open Access Journals (Sweden)

Ramachandran Vinoy K

2012-01-01

Full Text Available Abstract Background Invasion of intestinal epithelial cells by Salmonella enterica serovar Typhimurium (S. Typhimurium requires expression of the extracellular virulence gene expression programme (STEX, activation of which is dependent on the signalling molecule guanosine tetraphosphate (ppGpp. Recently, next-generation transcriptomics (RNA-seq has revealed the unexpected complexity of bacterial transcriptomes and in this report we use differential RNA sequencing (dRNA-seq to define the high-resolution transcriptomic architecture of wild-type S. Typhimurium and a ppGpp null strain under growth conditions which model STEX. In doing so we show that ppGpp plays a much wider role in regulating the S. Typhimurium STEX primary transcriptome than previously recognised. Results Here we report the precise mapping of transcriptional start sites (TSSs for 78% of the S. Typhimurium open reading frames (ORFs. The TSS mapping enabled a genome-wide promoter analysis resulting in the prediction of 169 alternative sigma factor binding sites, and the prediction of the structure of 625 operons. We also report the discovery of 55 new candidate small RNAs (sRNAs and 302 candidate antisense RNAs (asRNAs. We discovered 32 ppGpp-dependent alternative TSSs and determined the extent and level of ppGpp-dependent coding and non-coding transcription. We found that 34% and 20% of coding and non-coding RNA transcription respectively was ppGpp-dependent under these growth conditions, adding a further dimension to the role of this remarkable small regulatory molecule in enabling rapid adaptation to the infective environment. Conclusions The transcriptional architecture of S. Typhimurium and finer definition of the key role ppGpp plays in regulating Salmonella coding and non-coding transcription should promote the understanding of gene regulation in this important food borne pathogen and act as a resource for future research.

Antimicrobial resistance patterns of bovine Salmonella enterica isolates submitted to the Wisconsin Veterinary Diagnostic Laboratory: 2006-2015.

Science.gov (United States)

Valenzuela, J R; Sethi, A K; Aulik, N A; Poulsen, K P

2017-02-01

Salmonellosis on the dairy continues to have a significant effect on animal health and productivity and in the United States. Additionally, Salmonella enterica ssp. enterica causes an estimated 1.2 million cases of human illness annually. Contributing to the morbidity and mortality in both human and domestic animal species is emergence of antimicrobial resistance by Salmonella species and increased incidence of multidrug-resistant isolates. This study describes serotype distribution and the antimicrobial resistance patterns for various Salmonella serotypes isolated from bovine samples submitted to the Wisconsin Veterinary Diagnostic Laboratory (WVDL) over the past 10 yr. Salmonella serotyping and antimicrobial susceptibility testing data were obtained from the laboratory information management system at WVDL. Data from accessions were limited to bovine samples submitted to the WVDL between January 2006 and June 2015 and those that had both a definitive serotype and complete results for antimicrobial susceptibility testing. A total of 4,976 isolates were identified. Salmonella enterica ser. Dublin was the most prevalent serotype identified among bovine samples submitted to the WVDL, accounting for a total of 1,153 isolates (23% of total isolates) over the study period. Along with Dublin, Salmonella enterica ser. Cerro (795, 16%), Newport (720, 14%), Montevideo (421, 8%), Kentucky (419, 8%), and Typhimurium (202, 4%) comprised the top 6 most commonly isolated serotypes during that time. Overall, resistance of bovine Salmonella isolates in the study population remained stable, although decreases in resistance were noted for gentamicin, neomycin, and trimethoprim sulfamethoxazole during the study period. All isolates remained susceptible to enrofloxacin. These data show that antimicrobial susceptibility for bovine Salmonella has changed in the population served by WVDL in the past 10 yr. This information is important for understanding Salmonella disease ecology in

Selected lactic acid-producing bacterial isolates with the capacity to reduce Salmonella translocation and virulence gene expression in chickens.

Directory of Open Access Journals (Sweden)

Xiaojian Yang

Full Text Available BACKGROUND: Probiotics have been used to control Salmonella colonization/infection in chickens. Yet the mechanisms of probiotic effects are not fully understood. This study has characterized our previously-selected lactic acid-producing bacterial (LAB isolates for controlling Salmonella infection in chickens, particularly the mechanism underlying the control. METHODOLOGY/PRINCIPAL FINDINGS: In vitro studies were conducted to characterize 14 LAB isolates for their tolerance to low pH (2.0 and high bile salt (0.3-1.5% and susceptibility to antibiotics. Three chicken infection trials were subsequently carried out to evaluate four of the isolates for reducing the burden of Salmonella enterica serovar Typhimurium in the broiler cecum. Chicks were gavaged with LAB cultures (10(6-7 CFU/chick or phosphate-buffered saline (PBS at 1 day of age followed by Salmonella challenge (10(4 CFU/chick next day. Samples of cecal digesta, spleen, and liver were examined for Salmonella counts on days 1, 3, or 4 post-challenge. Salmonella in the cecum from Trial 3 was also assessed for the expression of ten virulence genes located in its pathogenicity island-1 (SPI-1. These genes play a role in Salmonella intestinal invasion. Tested LAB isolates (individuals or mixed cultures were unable to lower Salmonella burden in the chicken cecum, but able to attenuate Salmonella infection in the spleen and liver. The LAB treatments also reduced almost all SPI-1 virulence gene expression (9 out of 10 in the chicken cecum, particularly at the low dose. In vitro treatment with the extracellular culture fluid from a LAB culture also down-regulated most SPI-1 virulence gene expression. CONCLUSIONS/SIGNIFICANCE: The possible correlation between attenuation of Salmonella infection in the chicken spleen and liver and reduction of Salmonella SPI-1 virulence gene expression in the chicken cecum by LAB isolates is a new observation. Suppression of Salmonella virulence gene expression in

Presence and persistence of Salmonella enterica serotype typhimurium in the phyllosphere and rhizosphere of spray-irrigated parsley.

Science.gov (United States)

Kisluk, Guy; Yaron, Sima

2012-06-01

Salmonella enterica is one of the major food-borne pathogens associated with ready-to-eat fresh foods. Although polluted water might be a significant source of contamination in the field, factors that influence the transfer of Salmonella from water to the crops are not well understood, especially under conditions of low pathogen levels in water. The aim of this study was to investigate the short- and long-term (1 h to 28 days) persistence of Salmonella enterica serotype Typhimurium in the phyllosphere and the rhizosphere of parsley following spray irrigation with contaminated water. Plate counting and quantitative real-time PCR (qRT-PCR)-based methods were implemented for the quantification. By applying qRT-PCR with enrichment, we were able to show that even irrigation with water containing as little as ∼300 CFU/ml resulted in the persistence of S. Typhimurium on the plants for 48 h. Irrigation with water containing 8.5 log CFU/ml resulted in persistence of the bacteria in the phyllosphere and the rhizosphere for at least 4 weeks, but the population steadily declined with a major reduction in bacterial counts, of ∼2 log CFU/g, during the first 2 days. Higher levels of Salmonella were detected in the phyllosphere when plants were irrigated during the night compared to irrigation during the morning and during winter compared to the other seasons. Further elucidation of the mechanisms underlying the transfer of Salmonella from contaminated water to crops, as well as its persistence over time, will enable the implementation of effective irrigation and control strategies.

The epidemiology of Salmonella infection of calves: the role of dealers.

Science.gov (United States)

Wray, C.; Todd, N.; McLaren, I.; Beedell, Y.; Rowe, B.

1990-01-01

Salmonellas were detected in the environment of 10 of the 12 calf dealers' premises studied. The cleaning and disinfection routines were often ineffective and salmonellas were isolated from 7.6% and 5.3% of the wall and floor samples before disinfection and 6.8% and 7.6% afterwards. Eight different salmonella serotypes were detected, of which the commonest were Salmonella typhimurium, predominantly phage type DT204C, and S. dublin. Plasmid profiles were used to fingerprint S. typhimurium DT204C and the results indicated that with the exception of one of the premises, prolonged salmonella-persistence in the environment was not occurring. Three separate epidemics of salmonellosis in calves were studied by use of plasmid profile analysis. The results illustrated the role of delers, and their subcontractors, in the dissemination of salmonellas. The study concludes with suggestions for methods to reduce the spread of salmonellas in the calf marketing chain. PMID:2209734

Modelling transfer of Salmonella Typhimurium DT104 during simulation of grinding of pork.

Science.gov (United States)

Møller, C O A; Nauta, M J; Christensen, B B; Dalgaard, P; Hansen, T B

2012-01-01

The aim of this study was to develop a model to predict cross-contamination of Salmonella during grinding of pork. Transfer rates of Salmonella were measured in three experiments, where between 10 and 20 kg meat was ground into 200-g portions. In each experiment, five pork slices of about 200 g per slice were inoculated with 8-9 log-units of Salmonella Typhimurium DT104 and used for building up the contamination in the grinder. Subsequently, Salmonella-free slices were ground and collected as samples of c. 200 g minced pork. Throughout the process, representative samples were quantitatively analysed for Salmonella. A model suggested by Nauta et al. (2005) predicting cross-contamination of Campylobacter in poultry processing and two modified versions of this model were tested. The present study observed a tailing phenomenon of transfer of Salmonella during a small-scale grinding process. It was, therefore, hypothesized that transfer occurred from two environmental matrices inside the grinder and a model was developed. The developed model satisfactorily predicted the observed concentrations of Salmonella during its cross-contamination in the grinding of up to 110 pork slices. The proposed model provides an important tool to examine the effect of cross-contamination in quantitative microbial risk assessments and might also be applied to various other food processes where cross-contamination is involved. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

Use of a recombinant Salmonella enterica serovar Typhimurium strain expressing C-Raf for protection against C-Raf induced lung adenoma in mice

International Nuclear Information System (INIS)

Gentschev, Ivaylo; Fensterle, Joachim; Schmidt, Andreas; Potapenko, Tamara; Troppmair, Jakob; Goebel, Werner; Rapp, Ulf R

2005-01-01

Serine-threonine kinases of the Raf family (A-Raf, B-Raf, C-Raf) are central players in cellular signal transduction, and thus often causally involved in the development of cancer when mutated or over-expressed. Therefore these proteins are potential targets for immunotherapy and a possible basis for vaccine development against tumors. In this study we analyzed the functionality of a new live C-Raf vaccine based on an attenuated Salmonella enterica serovar Typhimurium aroA strain in two Raf dependent lung tumor mouse models. The antigen C-Raf has been fused to the C-terminal secretion signal of Escherichia coli α-hemolysin and expressed in secreted form by an attenuated aroA Salmonella enterica serovar Typhimurium strain via the α-hemolysin secretion pathway. The effect of the immunization with this recombinant C-Raf strain on wild-type C57BL/6 or lung tumor bearing transgenic BxB mice was analyzed using western blot and FACS analysis as well as specific tumor growth assays. C-Raf antigen was successfully expressed in secreted form by an attenuated Salmonella enterica serovar Typhimurium aroA strain using the E. coli hemolysin secretion system. Immunization of wild-type C57BL/6 or tumor bearing mice provoked specific C-Raf antibody and T-cell responses. Most importantly, the vaccine strain significantly reduced tumor growth in two transgenic mouse models of Raf oncogene-induced lung adenomas. The combination of the C-Raf antigen, hemolysin secretion system and Salmonella enterica serovar Typhimurium could form the basis for a new generation of live bacterial vaccines for the treatment of Raf dependent human malignancies

Survival of Salmonella Typhimurium on soybean sprouts after treatment with gaseous chlorine dioxide and biocontrol Pseudomonas bacteria

Science.gov (United States)

Control of Salmonella Typhimurium on sprouts and minimally processed produce is crucial for food and consumer safety. The aim of this research was to assess natural microflora populations on soybean and evaluate the effects of gaseous chlorine dioxide (ClO2) and biocontrol Pseudomonas on the surviva...

Experimental Salmonella typhimurium infections in rats. II. Active and passive immunization as protection against a lethal bacterial dose

DEFF Research Database (Denmark)

Hougen, H P; Jensen, E T; Klausen, B

1990-01-01

Immunization against a lethal dose of Salmonella typhimurium was studied in athymic and thymus-bearing LEW rats. Active immunization was performed with formalin-killed whole cell vaccine or sublethal infection prior to the lethal infection. After vaccination with killed bacteria the euthymic...... from immunized thymus grafted animals provided only limited protective effect, and treatment with cells from athymic animals had no effect. The study shows that although isogeneic thymus-grafted nude rats become resistent to reinfection with S. typhimurium, only large doses of spleen cells from...

Antimicrobial susceptibility of Salmonella isolates from healthy pigs and chickens (2008-2011).

Science.gov (United States)

de Jong, Anno; Smet, Annemieke; Ludwig, Carolin; Stephan, Bernd; De Graef, Evelyne; Vanrobaeys, Mia; Haesebrouck, Freddy

2014-07-16

Using the agar dilution method, antimicrobial susceptibility to human-use antibiotics was determined among Belgian faecal Salmonella isolates from healthy pigs and broiler chickens. Both epidemiological cut-off values and clinical breakpoints were applied for interpretation of the results. Cephalosporin-resistant isolates were examined for the presence of genes encoding CTX-M, SHV, TEM and CMY β-lactamases. All isolates with decreased quinolone susceptibility were screened for plasmid-borne genes qnr, qepA and aac(6')-Ib-cr. In all, 368 Salmonella isolates were recovered from pigs and 452 from chickens. Clinical resistance to ciprofloxacin was absent in isolates of both host species, and was 1.9 and 13.1% to cefotaxime in pig and poultry isolates, respectively. Decreased susceptibility to cefotaxime amounted to 2.2 and 0.7%, whereas for ciprofloxacin this was 3.0 and 23.0% in pig and poultry isolates, respectively. Ciprofloxacin decreased susceptibility was limited to few serovars, mainly Paratyphi B. Multidrug resistance was markedly higher for pig isolates (39.7%) than for chicken isolates (17.3%). Sixty-six cefotaxime-resistant isolates, 59 from chickens and 7 from pigs, were phenotypically determined as ESBL/AmpC producers; predominantly Paratyphi B and Typhimurium serovars. BlaCTX-M (mostly blaCTXM-1, but also blaCTXM-2 and blaCTXM-9) and blaTEM-52 were the predominant ESBL genes. Only few isolates expressed SHV-12 or an AmpC enzyme (CMY-2). Isolates of four serovars carried qnr genes: Brandenburg and Llandof from pigs, both qnrS; Indiana and Paratyphi B from chickens with qnrB and qnrA. The latter isolate carried blaCTX-M-9 and was the only strain with a plasmid-borne quinolone resistance gene among the ESBL/AmpC producers. This Salmonella survey confirms that the ESBL/AmpC producers are particularly prevalent in chickens (12.8%), and much less in pigs (1.9%). A link between plasmid-borne quinolone resistance genes and ESBLs/AmpC was uncommon. Copyright �

Identification of potential drug targets in Salmonella enterica sv. Typhimurium using metabolic modelling and experimental validation

DEFF Research Database (Denmark)

Hartman, Hassan B.; Fell, David A.; Rossell, Sergio

2014-01-01

Salmonella enterica sv. Typhimurium is an established model organism for Gram-negative, intracellular pathogens. Owing to the rapid spread of resistance to antibiotics among this group of pathogens, new approaches to identify suitable target proteins are required. Based on the genome sequence of ...

In vitro characterization and in vivo properties of Salmonellae lytic bacteriophages isolated from free-range layers

Directory of Open Access Journals (Sweden)

L Fiorentin

2004-06-01

Full Text Available Occurrence of food poisoning related to Salmonella-contaminated eggs and chicken meat has been frequent in humans. Salmonella Enteritidis (SE and Salmonella Typhimurium (ST are included among the most important paratyphoid salmonellae associated with chicken meat and eggs. Elimination of Salmonella at the pre-harvest stage can play a significant role in preventing the introduction of this pathogen into the food chain and consequently in the reduction of food poisoning in humans. Bactericidal bacteriophages may provide a natural, nontoxic, feasible and non-expensive component of the multi-factorial approach for a pre-harvest control of Salmonella in poultry. Five bacteriophages lytic for SE PT4 and ST were obtained from 107 samples of feces of free-range layers in Brazil. All bacteriophages were characterized in vitro and in vivo, showing head and tail morphology and dsDNA as nucleic acids. Results of "in vivo" studies suggested that bacteriophages do not remain in Salmonella-free birds longer than one day, whereas they multiply in Salmonella-infected birds for longer periods. Besides, selection for phage-resistant SE PT4 did not seem to occur in the short term. Isolated bacteriophages will be investigated for their potential for pre-harvest biocontrol of SE PT4 in poultry.

Beneficial Effects of Sodium Phenylbutyrate Administration during Infection with Salmonella enterica Serovar Typhimurium.

Science.gov (United States)

Jellbauer, Stefan; Perez Lopez, Araceli; Behnsen, Judith; Gao, Nina; Nguyen, Thao; Murphy, Clodagh; Edwards, Robert A; Raffatellu, Manuela

2016-09-01

Sodium phenylbutyrate (PBA) is a derivative of the short-chain fatty acid butyrate and is approved for treatment of urea cycle disorders and progressive familial intrahepatic cholestasis type 2. Previously known functions include histone deacetylase inhibitor, endoplasmic reticulum stress inhibitor, ammonia sink, and chemical chaperone. Here, we show that PBA has a previously undiscovered protective role in host mucosal defense during infection. Administration of PBA to Taconic mice resulted in the increase of intestinal Lactobacillales and segmented filamentous bacteria (SFB), as well as an increase of interleukin 17 (IL-17) production by intestinal cells. This effect was not observed in Jackson Laboratory mice, which are not colonized with SFB. Because previous studies showed that IL-17 plays a protective role during infection with mucosal pathogens, we hypothesized that Taconic mice treated with PBA would be more resistant to infection with Salmonella enterica serovar Typhimurium (S Typhimurium). By using the streptomycin-treated mouse model, we found that Taconic mice treated with PBA exhibited significantly lower S Typhimurium intestinal colonization and dissemination to the reticuloendothelial system, as well as lower levels of inflammation. The lower levels of S Typhimurium gut colonization and intestinal inflammation were not observed in Jackson Laboratory mice. Although PBA had no direct effect on bacterial replication, its administration reduced S Typhimurium epithelial cell invasion and lowered the induction of the proinflammatory cytokine IL-23 in macrophage-like cells. These effects likely contributed to the better outcome of infection in PBA-treated mice. Overall, our results suggest that PBA induces changes in the microbiota and in the mucosal immune response that can be beneficial to the host during infection with S Typhimurium and possibly other enteric pathogens. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Experimental infection of one-day-old chicks with Salmonella Serotypes Previously isolated from poultry facilities, wild birds, and swine

Directory of Open Access Journals (Sweden)

E de Sousa

2013-12-01

Full Text Available In order to maintain the high production and export rates achieved by the Brazilian poultry industry, it is necessary to prevent and control certain disease agents, such as Salmonella spp. Using bacterial cultures, the aim of the present study was to investigate the prevalence of Salmonella spp. in specimens collected from broiler facilities. Local wild birds were also sampled, as well as the feces of swine housed on the poultry farm. After sample collection, the isolated serotypes were subsequently inoculated into broiler chicks to determine their effects. Positive samples were collected from the following locations in the poultry facilities: poultry litter (S. serotype 4,5,12:R:-; S. Heidelberg; S. Infantis, broiler feces (S. Heidelberg; S. serotype 6,7:R:-; S. serotype 4,5,12:R:-; S. Tennessee, water (S. Glostrup; S. serotype 6,8:d:-;, and lesser mealworms (Alphitobius diaperinus found in the litter (S. Tennessee. Among the 36 wild birds captured, S. Heidelberg was isolated from one bird's organs and intestinal contents (Colaptes campestris, and S. Enteritidis was isolated from another bird's intestinal contents (Zenaida auriculata. Salmonella Panama and Salmonella Typhimurium were isolated from swine feces. One-day-old chicks (150 were divided into 10 groups of 15 animals each. Each group was orally inoculated with a previously isolated serotype of Salmonella. Soft stools were observed on the cage floor and around the birds' cloaca between 3 and 12 days post-infection (dpi. The different serotypes of Salmonella used to inoculate the chicks were re-isolated from the spleen, liver, and cecal content samples of the infected birds on 15 and 21 dpi.

Structural basis for the mechanism of inhibition of uridine phosphorylase from Salmonella typhimurium

Energy Technology Data Exchange (ETDEWEB)

Lashkov, A. A.; Zhukhlistova, N. E.; Sotnichenko, S. E.; Gabdulkhakov, A. G.; Mikhailov, A. M., E-mail: amm@ns.crys.ras.ru [Russian Academy of Sciences, Shubnikov Institute of Crystallography (Russian Federation)

2010-01-15

The three-dimensional structures of three complexes of Salmonella typhimurium uridine phosphorylase with the inhibitor 2,2'-anhydrouridine, the substrate PO{sub 4}, and with both the inhibitor 2,2'-anhydrouridine and the substrate PO{sub 4} (a binary complex) were studied in detail by X-ray diffraction. The structures of the complexes were refined at 2.38, 1.5, and 1.75 A resolution, respectively. Changes in the three-dimensional structure of the subunits in different crystal structures are considered depending on the presence or absence of the inhibitor molecule and (or) the phosphate ion in the active site of the enzyme. The presence of the phosphate ion in the phosphate-binding site was found to substantially change the orientations of the side chains of the amino-acid residues Arg30, Arg91, and Arg48 coordinated to this ion. A comparison showed that the highly flexible loop L9 is unstable. The atomic coordinates of the refined structures of the complexes and the corresponding structure factors were deposited in the Protein Data Bank (their PDB ID codes are 3DD0 and 3C74). The experimental data on the spatial reorganization of the active site caused by changes in its functional state from the unligated to the completely inhibited state suggest the structural basis for the mechanism of inhibition of Salmonella typhimurium uridine phosphorylase.

Effect of ionizing radiation dose, temperature, and atmosphere on the survival of Salmonella typhimurium in sterile, mechanically deboned chicken meat

International Nuclear Information System (INIS)

Thayer, D.W.; Boyd, G.

1991-01-01

The response to gamma radiation (0 to 3.60 kGy; 100 krad = 1 kGy) of Salmonella typhimurium was tested in otherwise sterile, mechanically deboned chicken meat (MDCM) in the absence of competing microflora. Response was determined at temperatures of -20 to +20 C and when the MDCM was packaged in vacuum or in the presence of air. A central composite response-surface design was used to test the response of the pathogen to the treatments in a single experiment. Predictive equations were developed from the analyses of variances of the resulting data. The accuracy of each predictive equation was tested by further studies of the effects of gamma radiation on S. typhimurium in the presence or absence of air at -20, 0, and +20 C. All data were then analyzed to refine the predictive equations further. Both the original and the refined equations adequately predicted the response of S. typhimurium in MDCM to gamma radiation doses up to 3.60 kGy in the presence of air or in vacuo. Gamma irradiation was significantly more lethal for S. typhimurium in the presence of air and at higher temperatures. The final equations predict a reduction in the number of surviving Salmonella in MDCM irradiated to 1.50 kGy at -20 C of 2.53 logs in air or 2.12 logs if irradiated in vacuum. If the contaminated MDCM were to receive a dose of 3.0 kGy at -20 C in air, the number of Salmonella would be decreased by 4.78 logs, and if irradiated in vacuum, by 4.29 logs

Sobrevivência e perfil de resistência a antimicrobianos de Salmonella sp. isoladas em um sistema de tratamento de dejetos de suínos Survival and resistance patterns of Salmonella sp. isolated in a pig slurry treatment plant

Directory of Open Access Journals (Sweden)

Verônica Schmidt

2003-10-01

Full Text Available No presente estudo, foi avaliada a sobrevivência de Salmonella sp., presente em dejetos suínos, durante tratamento em um sistema de separação física e lagoas de estabilização ligadas em série. Nas amostras de Salmonella sp. isoladas foi determinado o perfil de resistência pelo método de difusão em ágar, usando 14 antimicrobianos. Das 20 coletas realizadas, foi possível isolar Salmonella sp. em 13 coletas no ponto correspondente ao início do sistema de tratamento e em apenas uma no ponto final do mesmo. Amostras de Salmonella sp. isoladas (161/163 pertenciam ao sorotipo Typhimurium e demonstraram resistência contra sulfonamida (100%, tetraciclina (99,4%, estreptomicina (90,1%, sulfa/trimetoprima (84,5%, ácido nalidíxico (77,6%, ampicilina (76,4%, cloranfenicol (29,2%, cefaclor (25,5%, tobramicina (13,7%, gentamicina (6,2%, amoxacilina/ácido clavulânico (5%, neomicina (5% e amicacina (3,7%. A maioria (94,5% das amostras isoladas foram resistente a 4 ou mais antimicrobianos e apresentaram grande variabilidade nos perfis de resistência. O nível de resistência e a variabilidade dos perfis mantiveram-se em nível semelhante ao longo do sistema.The survival of Salmonella sp. in pig slurry submitted to treatment in successive stabilization ponds on a pig-breeding farm was investigated. Furthermore, the isolated Salmonella strains were tested for their resistance against 14 antibiotics, using the agar diffusion method. Of a total of 20 samples taken from different points in the stabilization ponds system, 13 were positive for Salmonella sp. in the beginning and only one at the end of the system. Most of the isolated Salmonella strains (161/163 belonged to sorovar Typhimurium. These strains were resistant to sulfonamide (100%, tetracycline (9.4%, sulfamethoxazole/trimethoprin (84.5%, ampicillin (76.4%, cloramphenicol (29.2%, streptomycin (90.1%, nalidixic acid (77.6%, tobramycin (13.7%, neomycin (5%, amikacin (3.7%, cefaclor (25

Effect of Pulsed Electric Field on Membrane Lipids and Oxidative Injury of Salmonella typhimurium.

Science.gov (United States)

Yun, Ou; Zeng, Xin-An; Brennan, Charles S; Han, Zhong

2016-08-22

Salmonella typhimurium cells were subjected to pulsed electric field (PEF) treatment at 25 kV/cm for 0-4 ms to investigate the effect of PEF on the cytoplasmic membrane lipids and oxidative injury of cells. Results indicated that PEF treatment induced a decrease of membrane fluidity of Salmonella typhimurium (S. typhimuriumi), possibly due to the alterations of fatty acid biosynthesis-associated gene expressions (down-regulation of cfa and fabA gene expressions and the up-regulation of fabD gene expression), which, in turn, modified the composition of membrane lipid (decrease in the content ratio of unsaturated fatty acids to saturated fatty acids). In addition, oxidative injury induced by PEF treatment was associated with an increase in the content of malondialdehyde. The up-regulation of cytochrome bo oxidase gene expressions (cyoA, cyoB, and cyoC) indicated that membrane damage was induced by PEF treatment, which was related to the repairing mechanism of alleviating the oxidative injury caused by PEF treatment. Based on these results, we achieved better understanding of microbial injury induced by PEF, suggesting that micro-organisms tend to decrease membrane fluidity in response to PEF treatment and, thus, a greater membrane fluidity might improve the efficiency of PEF treatment to inactivate micro-organisms.

Functional and crystallographic characterization of Salmonella typhimurium Cu,Zn superoxide dismutase coded by the sodCI virulence gene

NARCIS (Netherlands)

Pesce, A; Battistoni, A; Stroppolo, ME; Polizio, F; Nardini, M; Kroll, JS; Langford, PR; O'Neill, P; Sette, M; Desideri, A; Bolognesi, M

2000-01-01

The functional and three-dimensional structural features of Cu,Zn superoxide dismutase coded by the Salmonella typhimurium sodCI gene, have been characterized. Measurements of the catalytic rate indicate that this enzyme is the most efficient superoxide dismutase analyzed so far, a feature that may

Genotypic homogeneity of multidrug resistant S. Typhimurium infecting distinct adult and childhood susceptibility groups in Blantyre, Malawi.

Directory of Open Access Journals (Sweden)

Chisomo L Msefula

Full Text Available Nontyphoidal Salmonella (NTS serovars are a common cause of bacteraemia in young children and HIV-infected adults in Malawi and elsewhere in sub-Saharan Africa. These patient populations provide diverse host-immune environments that have the potential to drive bacterial adaptation and evolution. We therefore investigated the diversity of 27 multidrug resistant (MDR Salmonella Typhimurium strains isolated over 6 years (2002-2008 from HIV-infected adults and children and HIV-uninfected children. Sequence reads from whole-genome sequencing of these isolates using the Illumina GA platform were mapped to the genome of the laboratory strain S. Typhimurium SL1344 excluding homoplastic regions that contained prophage and insertion elements. A phylogenetic tree generated from single nucleotide polymorphisms showed that all 27 strains clustered with the prototypical MDR strain D23580. There was no clustering of strains based on host HIV status or age, suggesting that these susceptible populations acquire S. Typhimurium from common sources or that isolates are transmitted freely between these populations. However, 7/14 of the most recent isolates (2006/2008 formed a distinct clade that branched off 22 SNPs away from the cluster containing earlier isolates. These data suggest that the MDR bacterial population is not static, but is undergoing microevolution which might result in further epidemiology change.

Sewage sludge amendment and inoculation with plant-parasitic nematodes do not facilitate the internalization of Salmonella Typhimurium LT2 in lettuce plants.

Science.gov (United States)

Fornefeld, Eva; Baklawa, Mohamed; Hallmann, Johannes; Schikora, Adam; Smalla, Kornelia

2018-05-01

Contamination of fruits and vegetables with Salmonella is a serious threat to human health. In order to prevent possible contaminations of fresh produce it is necessary to identify the contributing ecological factors. In this study we investigated whether the addition of sewage sludge or the presence of plant-parasitic nematodes foster the internalization of Salmonella enterica serovar Typhimurium LT2 into lettuce plants, posing a potential threat for human health. Greenhouse experiments were conducted to investigate whether the amendment of sewage sludge to soil or the presence of plant-parasitic nematodes Meloidogyne hapla or Pratylenchus crenatus promote the internalization of S. Typhimurium LT2 from soil into the edible part of lettuce plants. Unexpectedly, numbers of cultivable S. Typhimurium LT2 decreased faster in soil with sewage sludge than in control soil but not in root samples. Denaturing gradient gel electrophoresis analysis revealed shifts of the soil bacterial communities in response to sewage sludge amendment and time. Infection and proliferation of nematodes inside plant roots were observed but did not influence the number of cultivable S. Typhimurium LT2 in the root samples or in soil. S. Typhimurium LT2 was not detected in the leaf samples 21 and 49 days after inoculation. The results indicate that addition of sewage sludge, M. hapla or P. crenatus to soil inoculated with S. Typhimurium LT2 did not result in an improved survival in soil or internalization of lettuce plants. Copyright © 2017. Published by Elsevier Ltd.

Gold nanoparticles enhanced SERS aptasensor for the simultaneous detection of Salmonella typhimurium and Staphylococcus aureus.

Science.gov (United States)

Zhang, Hui; Ma, Xiaoyuan; Liu, Ying; Duan, Nuo; Wu, Shijia; Wang, Zhouping; Xu, Baocai

2015-12-15

Salmonella typhimurium and Staphylococcus aureus are most common causes of food-associated disease. A Raman based biosensor was developed for S. typhimurium and S. aureus detection simultaneously. The biosensor was based on nanoparticles enhanced Raman intensity and the specific recognition of aptamer. The Raman signal probe and the capture probe are built. Gold nanoparticles (GNPs) modified with Raman molecules (Mercaptobenzoic acid and 5,5'-Dithiobis(2-nitrobenzoic acid)) and aptamer are used as the signal probe for S. typhimurium and S. aureus, respectively. Fe3O4 magnetic gold nanoparticles (MGNPs) immobilized with both aptamer of S. typhimurium and S. aureus are used as the capture probe. When S. typhimurium and S. aureus are added in the reaction system, the capture probe will capture the target bacteria through the specific binding effect of aptamer. And then the signal probe will be connected to the bacteria also by the effect of aptamer to form the sandwich like detection structure. The Raman intensified spectrum was measured to quantify S. typhimurium and S. aureus. Under optimal conditions, the SERS intensity of MBA at 1582 cm(-1) are used to measure S. typhimurium (y=186.4762+704.8571x, R(2)=0.9921) and the SERS intensity of DNTB at 1333 cm(-1) are used to measure S. aureus (y=135.2381+211.4286x, R(2)=0.9946) in the range of 10(2)-10(7) cfu mL(-1). The LOD is 35 cfu mL(-1) for S. aureus and 15 cfu mL(-1) for S. typhimurium. This method is simple and rapid, results in high sensitivity and specificity, and can be used to detect actual samples. Copyright © 2015 Elsevier B.V. All rights reserved.

A rapid and specific detection of pathogenic serovar Salmonella typhimurium by loop-mediated isothermal amplification method (LAMP

Directory of Open Access Journals (Sweden)

Hadi Ravan

2017-09-01

Discussion and conclusion: As a result of a high sensitivity and specificity of the method as well as its low cost per assay, it could be concluded that the present LAMP assay is a powerful, accurate, and efficient method for detecting pathogenic serovar Salmonella typhimurium in food-processing industries and diagnostic laboratories.

Caracterização de sorotipos em linhagens do gênero Salmonella isoladas de diferentes afecções em animais domésticos

Directory of Open Access Journals (Sweden)

Márcio G. Ribeiro

2010-02-01

Full Text Available Foram caracterizados os sorotipos, o perfil de sensibilidade microbiana e os achados clínico-epidemiológicos em 53 linhagens do gênero Salmonella isoladas de 41 cães, nove equinos e três bovinos, acometidos por diferentes manifestações clínicas entre 1997 e 2007. Salmonella Typhimurium (45,3%, Salmonella enterica (22,6%, Salmonella Enteritidis (7,5%, Salmonella enterica subsp enterica 4,5,12i (5,7%, Salmonella Newport (5,7%, Salmonella Dublin (3,8%, Salmonella Agona (3,8%, Salmonella Glostrup (3,8%, Salmonella Saintpaul (1,8% foram os sorotipos encontrados. Ciprofloxacina (100,0%, norfloxacina (100,0% e gentamicina (100,0% foram os antimicrobianos mais efetivos, enquanto a maior resistência das linhagens foi observada para ceftiofur (28,5% e florfenicol (7,0%. As linhagens foram isoladas de animais com enterite, infecção do trato urinário, septicemia, piometra, pneumonia e conjuntivite. Ressalta-se para o predomínio do sorovar Typhimurium nas diferentes manifestações da salmonelose nos animais. Destaca-se, também, a identificação de sorotipos nos animais que também são observados em casos de salmonelose em humanosThe serotype characterization, antimicrobial susceptibility profile, and clinical-epidemiological findings were evaluated in 53 Salmonella spp. strains isolated from 41 dogs, nine horses and three cattle presenting different clinical manifestations between 1997 at 2007. Salmonella Typhimurium (45.3%, Salmonella enterica (22.6%, Salmonella Enteritidis (7.5%, Salmonella enterica subsp. enterica 4,5,12i (5.7%, Salmonella Newport (5.7%, Salmonella Dublin (3.8%, Salmonella Agona (3.8%, Salmonella Glostrup (3.8%, Salmonella Saintpaul (1.8% were the more common serotypes. Ciprofloxacin (100.0%, norfloxacin (100.0% and gentamicin (100.0% were more effective drugs while resistance of isolates was observed to ceftiofur (28.5% and florfenicol (7.0%. The strains were isolated from animals with enteritis, urinary tract infections

Effect of the oyster contaminated in Salmonella typhimurium and Escherichia coli

International Nuclear Information System (INIS)

Brena V, M.

1992-08-01

In this work the effect of the oyster contaminated in the species of bacteria better studied by its genetic composition these are the Salmonella typhimurium and the Escherichia coli and that its have been starting point for later radiobiological studies in superior organisms. Of this its have arisen two general lines. The research about the genotoxic effect of substances or mixtures in bacteria with the collaboration of the groups of Drosophila and X-ray Fluorescence analysis and on the other hand the study of the low doses of radiation also in bacteria. It is also treated the topic about the genetic effect of aromatic hydrocarbons in different biological systems. (Author)

Sonication reduces the attachment of Salmonella Typhimurium ATCC 14028 cells to bacterial cellulose-based plant cell wall models and cut plant material.

Science.gov (United States)

Tan, Michelle S F; Rahman, Sadequr; Dykes, Gary A

2017-04-01

This study investigated the removal of bacterial surface structures, particularly flagella, using sonication, and examined its effect on the attachment of Salmonella Typhimurium ATCC 14028 cells to plant cell walls. S. Typhimurium ATCC 14028 cells were subjected to sonication at 20 kHz to remove surface structures without affecting cell viability. Effective removal of flagella was determined by staining flagella of sonicated cells with Ryu's stain and enumerating the flagella remaining by direct microscopic counting. The attachment of sonicated S. Typhimurium cells to bacterial cellulose-based plant cell wall models and cut plant material (potato, apple, lettuce) was then evaluated. Varying concentrations of pectin and/or xyloglucan were used to produce a range of bacterial cellulose-based plant cell wall models. As compared to the non-sonicated controls, sonicated S. Typhimurium cells attached in significantly lower numbers (between 0.5 and 1.0 log CFU/cm 2 ) to all surfaces except to the bacterial cellulose-only composite without pectin and xyloglucan. Since attachment of S. Typhimurium to the bacterial cellulose-only composite was not affected by sonication, this suggests that bacterial surface structures, particularly flagella, could have specific interactions with pectin and xyloglucan. This study indicates that sonication may have potential applications for reducing Salmonella attachment during the processing of fresh produce. Copyright © 2016 Elsevier Ltd. All rights reserved.

Characterization of Salmonella Typhimurium isolates from domestically acquired infections in Finland by phage typing, antimicrobial susceptibility testing, PFGE and MLVA.

Science.gov (United States)

Lienemann, Taru; Kyyhkynen, Aino; Halkilahti, Jani; Haukka, Kaisa; Siitonen, Anja

2015-07-02

Salmonella enterica spp. enterica serotype Typhimurium (STM) is the most common agent of domestically acquired salmonellosis in Finland. Subtyping methods which allow the characterization of STM are essential for effective laboratory-based STM surveillance and for recognition of outbreaks. This study describes the diversity of Finnish STM isolates using phage typing, antimicrobial susceptible testing, pulsed-field gel electrophoresis (PFGE) and multilocus variable-number tandem repeat analysis (MLVA), and compares the discriminatory power and the concordance of these methods. A total of 375 sporadic STM isolates were analysed. The isolates were divided into 31 definite phage (DT) types, dominated by DT1 (47 % of the isolates), U277 (9 % of the isolates) and DT104 (8 % of the isolates). Of all the isolates, 62 % were susceptible to all the 12 antimicrobials tested and 11 % were multidrug resistant. Subtyping resulted in 83 different XbaI-PFGE profiles and 111 MLVA types. The three most common XbaI-PFGE profiles (STYM1, STYM7 and STYM8) and one MLVA profile with three single locus variants accounted for 56 % and 49 % of the STM isolates, respectively. The studied isolates showed a genetic similarity of more than 70 % by XbaI-PFGE. In MLVA, 71 % of the isolates lacked STTR6 and 77 % missed STTR10p loci. Nevertheless, the calculated Simpson's diversity index for XbaI-PFGE was 0.829 (95 % CI 0.792-0.865) and for MLVA 0.867 (95 % CI 0.835-0.898). However, the discriminatory power of the 5-loci MLVA varied among the phage types. The highest concordance of the results was found between XbaI-PFGE and phage typing (adjusted Wallace coefficient was 0.833 and adjusted Rand coefficient was 0.627). In general, the calculated discriminatory power was higher for genotyping methods (MLVA and XbaI-PFGE) than for phenotyping methods (phage typing). Overall, comparable diversity indices were calculated for PFGE and MLVA (both DI > 0.8). However, MLVA was phage type dependent

The common structural architecture of Shigella flexneri and Salmonella typhimurium type three secretion needles.

Directory of Open Access Journals (Sweden)

Jean-Philippe Demers

2013-03-01

Full Text Available The Type Three Secretion System (T3SS, or injectisome, is a macromolecular infection machinery present in many pathogenic Gram-negative bacteria. It consists of a basal body, anchored in both bacterial membranes, and a hollow needle through which effector proteins are delivered into the target host cell. Two different architectures of the T3SS needle have been previously proposed. First, an atomic model of the Salmonella typhimurium needle was generated from solid-state NMR data. The needle subunit protein, PrgI, comprises a rigid-extended N-terminal segment and a helix-loop-helix motif with the N-terminus located on the outside face of the needle. Second, a model of the Shigella flexneri needle was generated from a high-resolution 7.7-Å cryo-electron microscopy density map. The subunit protein, MxiH, contains an N-terminal α-helix, a loop, another α-helix, a 14-residue-long β-hairpin (Q51-Q64 and a C-terminal α-helix, with the N-terminus facing inward to the lumen of the needle. In the current study, we carried out solid-state NMR measurements of wild-type Shigella flexneri needles polymerized in vitro and identified the following secondary structure elements for MxiH: a rigid-extended N-terminal segment (S2-T11, an α-helix (L12-A38, a loop (E39-P44 and a C-terminal α-helix (Q45-R83. Using immunogold labeling in vitro and in vivo on functional needles, we located the N-terminus of MxiH subunits on the exterior of the assembly, consistent with evolutionary sequence conservation patterns and mutagenesis data. We generated a homology model of Shigella flexneri needles compatible with both experimental data: the MxiH solid-state NMR chemical shifts and the state-of-the-art cryoEM density map. These results corroborate the solid-state NMR structure previously solved for Salmonella typhimurium PrgI needles and establish that Shigella flexneri and Salmonella typhimurium subunit proteins adopt a conserved structure and orientation in their

Contaminated Larval and Adult Lesser Mealworms, Alphitobius diaperinus (Coleoptera: Tenebrionidae)can Transmit Salmonella Typhimurium in a Broiler Flock

Science.gov (United States)

The ability of the lesser mealworm, Alphitobius diaperinus (Panzer), commonly known as the darkling beetle, to transmit a marker strain Salmonella Typhimurium to day-of-hatch broiler chicks was evaluated, as well as the spread to non-challenged pen mates. Day-of-hatch chicks were orally gavaged wit...

Association with MDCK epithelial cells by Salmonella typhimurium is reduced during utilization of carbohydrates.

OpenAIRE

Schiemann, D A

1995-01-01

Association of Salmonella typhimurium with MDCK epithelial cells in monolayers, represented primarily by intracellular bacteria after 30 min of contact, with centrifugation followed by vigorous washing, was measured during aerobic and anaerobic growth of the bacteria in brain heart infusion broth. Cell association was greatest during a short period in the late log phase of growth under aerobic conditions. At this time, the pH of the growth medium was changing from acid to alkaline and glucose...

Combination of peracetic acid and ultrasound reduces Salmonella Typhimurium on fresh lettuce (Lactuca sativa L. var. crispa).

Science.gov (United States)

Silveira, Luiza Oliveira; do Rosário, Denes Kaic Alves; Giori, Ana Carolina Garcia; Oliveira, Syllas Borburema Silva; da Silva Mutz, Yhan; Marques, Clara Suprani; Coelho, Jussara Moreira; Bernardes, Patrícia Campos

2018-04-01

Salmonella outbreaks related to fruits and vegetables have been reported being lettuce one of the most contaminated. Peracetic acid (PA) at 50 mg/L, sodium dichloroisocyanurate (SD) at 100 mg/L, and the combination of SD at 100 mg/L and babaçu coconut ( Attalea speciosa ) oil detergent at 100 mg/L were applied to fresh lettuce. Natural contaminant microbiota, physicochemical characteristics, and sensory attributes were evaluated. PA and SD reduced mesophilic aerobic counts by 2.1 and 1.5 log cfu/g, respectively. The most efficient treatment in reducing natural microbiota (i.e., PA) was applied alone and in combination with ultrasound (US). It reduced Salmonella enterica Typhimurium counts to undetectable levels (< 1 log cfu/g). US further reduced S. Typhimurium counts by 0.6 log cfu/g in relation to PA, treatment which lessened the pH but increased the titratable acidity of lettuce, but did not cause total color difference. Therefore, the combination of PA and US holds a potential industrial application for sanitization purposes.

In vitro quantitative analysis of Salmonella typhimurium preference for amino acids secreted by human breast tumor

Science.gov (United States)

Choi, Eunpyo; Maeng, Bohee; Lee, Jae-hun; Chang, Hyung-kwan; Park, Jungyul

2016-12-01

Bacterial therapies have been paid significant attentions by their ability to penetrate deep into the solid tumor tissue and its propensity to naturally accumulate in tumors of living animals. Understanding the actual mechanism for bacteria to target the tumor is therapeutically crucial but is poorly understood. We hypothesized that amino acids released from the specific tumors induced bacteria to those tumors and the experiments for chemotactic response of bacteria toward the cancer secreting amino acids was then performed by using the diffusion based multiple chemical gradient generator constructed by in situ self-assembly of microspheres. The quantitative analysis was carried out by comparison of intensity using green fluorescent protein (GFP) tagged Salmonella typhimurium ( S. typhimurium) in the gradient generator, which showed the clear preference to the released amino acids, especially from breast cancer patients. The understanding chemotaxis toward the cancer secreting amino acids is essential for controlling S. typhimurium targeting in tumors and will allow for the development of bacterial therapies.

Radiation-induced mutagenicity and lethality in Salmonella typhimurium

International Nuclear Information System (INIS)

Isildar, M.; Bakale, G.

1983-01-01

The mutagenic and lethal effects of ionizing radiation on histidine-deficient auxotrophs of Salmonella typhimurium were studied to improve the understanding of radiation damage to DNA. The auxotrophs were divided into two groups - one which is sensitive to base-pair substitutions and another sensitive to frameshifts. These groups were composed of parent-daughter pairs in which the chemical mutagenicity enhancing plasmid, pKM101, is absent in the parent strain and present in the daughter. Co-60 #betta#-radiation and 250 kV x-rays were used to irradiate the bacteria. Irradiation of the frameshift - sensitive strains which carry the pKm101 plasmid doubled the absolute number of induced revertants whereas irradiation of the base-pair substitution sensitive strain which also carries the pKm101 plasmid produced nearly no change in the number of induced revertants. A nearly negligible effect on the mutation rate was observed for all parent strains

The pharmacodynamic effect of amoxicillin and danofloxacin against Salmonella typhimurium in an in-vitro pharmacodynamic model

DEFF Research Database (Denmark)

Lindecrona, R.H.; Friis, C.; Aarestrup, Frank Møller

2000-01-01

The pharmacodynamic effect of amoxicillin and danofloxacin against two strains of Salmonella typhimurium was examined in an in-vitro pharmacodynamic model. For amoxicillin, peak concentrations of 1, 2 and 4 mu g ml(-1) and half-lives (t(1/2) of 3 and 15 hours were evaluated. For danofloxacin peak...... concentrations of 0.25, 0.50 and 1.50 mu g ml(-1) and half-lives of 7 and 15 hours were examined. For amoxicillin both the peak concentration and the half-life influenced the pharmacodynamic effect (P pharmacodynamic effect was observed when the antibiotic concentration was greater than minimum...... inhibitory concentration for 79 per cent or more of the dosing interval. The hires of the isolates increased when the amoxicillin concentrations were close to the nac during the first hours of exposure. For danofloxacin the pharmacodynamic effect was dependent on the peak concentration only (P

Emergence of Salmonella genomic island 1 (SGI1) among Proteus mirabilis clinical isolates in Dijon, France.

Science.gov (United States)

Siebor, Eliane; Neuwirth, Catherine

2013-08-01

Salmonella genomic island 1 (SGI1) is often encountered in antibiotic-resistant Salmonella enterica and exceptionally in Proteus mirabilis. We investigated the prevalence of SGI1-producing clinical isolates of P. mirabilis in our hospital (Dijon, France). A total of 57 strains of P. mirabilis resistant to amoxicillin and/or gentamicin and/or trimethoprim/sulfamethoxazole isolated from August 2011 to February 2012 as well as 9 extended-spectrum β-lactamase (ESBL)-producing P. mirabilis from our collection were tested for the presence of SGI1 by PCR. The complete SGI1 structure from positive isolates [backbone and multidrug resistance (MDR) region] was sequenced. SGI1 was detected in 7 isolates; 5 out of the 57 isolates collected during the study period (9%) and 2 out of the 9 ESBL-producing strains of our collection. The structures of the seven SGI1s were distinct. Three different backbones were identified: one identical to the SGI1 backbone from the epidemic Salmonella Typhimurium DT104, one with variations already described in SGI1-K from Salmonella Kentucky (deletion and insertion of IS1359 in the region spanning from S005 to S009) and one with a variation never detected before (deletion from S005 to S009). Six different MDR regions were identified: four simple variants containing resistance genes already described and two variants harbouring a very complex structure including regions derived from several transposons and IS26 elements with aphA1a never reported to date in SGI1. SGI1 variants are widely distributed among P. mirabilis clinical strains and might spread to other commensal Enterobacteriaceae. This would become a serious public health problem.

Assessing the prevalence of Salmonella enterica in poultry hatcheries by using hatched eggshell membranes.

Science.gov (United States)

Chao, M-R; Hsien, C-H; Yeh, C-M; Chou, S-J; Chu, C; Su, Y-C; Yu, C-Y

2007-08-01

Salmonella enterica causes a number of significant poultry diseases and is also a major pathogen in humans. Most poultry infected by Salmonella become carriers; infection may also be fatal, depending on the particular serovar and the age of the bird at infection. Younger birds are more susceptible to infection by Salmonella, so it is critical that hatcheries monitor birds. We developed a method to use hatched eggshell membranes (HEM) to assess contamination by Salmonella in poultry hatching cabinets and to evaluate the prevalence of Salmonella in a goose hatchery and rearing farm. Comparison of the Salmonella isolation rate in hatching cabinets using 3 sampling methods showed that the highest Salmonella contamination was detected in HEM, and that these results differed significantly from those obtained from fluff samples and cabinet swab samples (P chicken, and duck hatcheries. The lowest Salmonella-positive rate was found for the chicken hatchery, followed by the goose and the duck hatcheries (P hatcheries: A, B, C1, C2, D, and E. The distribution of these serogroups differed among the hatcheries. Salmonella serogroup C1 was the major serogroup found in geese, compared with serogroup B in chickens and ducks. However, Salmonella Typhimurium was dominant in 1 goose hatchery and also in geese from this hatchery that had been transferred to a farm. Antibiotic susceptibility analysis showed that Salmonella Typhimurium strains isolated from the farm geese with diarrhea showed significantly higher resistance to doxycycline, colistin, sulfamethoxazole-trimethoprin, and cephalothin than those isolated from the hatchery (P hatcheries and rearing farms.

Bovine salmonellosis in Northeast of Iran: Frequency, genetic fingerprinting and antimicrobial resistance patterns of Salmonella spp.

Directory of Open Access Journals (Sweden)

Hessam A. Halimi

2014-01-01

Conclusion: The emergence of multiple antibiotic-resistant strains of Salmonella Typhimurium should be of great concern to the public. No correlation between ERIC fingerprinting and resistance patterns of Salmonella isolates was found, which indicates resistance to antimicrobial agents was not related to specific genetic background.

Impact of Clinical Salmonellosis in Veal Calves on the Recovery of Salmonella in Lymph Nodes at Harvest.

Science.gov (United States)

Muñoz-Vargas, Lohendy; Finney, Sarah K; Hutchinson, Holden; Masterson, Margaret A; Habing, Greg

2017-11-01

The objective of this study was to determine the prevalence, serotypes, antimicrobial resistance phenotypes, and pulsed-field gel electrophoresis (PFGE) patterns of Salmonella recovered in feces and mesenteric and prefemoral lymph nodes (LNs) from cohorts of calves with and without a confirmed outbreak of salmonellosis. In a prospective cohort study, 160 calves from four farms without a reported outbreak (nonoutbreak farms) were sampled at farm and harvest. In addition, harvest samples from 80 calves of two farms with a confirmed outbreak (outbreak farms) were collected. A culture protocol for Salmonella isolation was applied for all samples and recovered isolates were further characterized by serotyping, antimicrobial susceptibility testing, and PFGE. Among nonoutbreak farms, Salmonella was recovered from 0% (0/160) farm fecal samples, 3.7% (6/160) harvest fecal swabs, 21.9% (35/160) mesenteric LNs, and 0.6% (1/160) prefemoral LNs. Serotypes identified in nonoutbreak herds included Salmonella Typhimurium, Cerro, Hartford, and Newport. Most isolates (64.3%, 27/42) exhibited a unique multidrug-resistant (MDR) phenotype, including resistance to extended-spectrum cephalosporins. Salmonella prevalence in harvest fecal samples and prefemoral LNs among calves from outbreak farms was numerically higher, but not significantly different than those without an outbreak. Serotypes recovered from outbreak farms included Salmonella Heidelberg and Typhimurium, and the monophasic Salmonella Typhimurium strains 4,5,12:i:- and 4,12:i:-, which have been also reported as highly pathogenic in humans. All isolates (33/33) exhibited an MDR phenotype. Salmonella strains recovered from ill calves in two outbreaks had indistinguishable PFGE patterns, suggesting between-farm transmission. In addition, the genotype of Salmonella Heidelberg causing an outbreak among calves was recovered from three prefemoral LNs of surviving members of the cohort at harvest. Implementation of preharvest

Prevalence, serotypes and resistance patterns of Salmonella in Danish pig production

DEFF Research Database (Denmark)

Arguello, Hector; Sørensen, Gitte; Carvajal, Ana

2013-01-01

Typhimurium in finishers and Salmonella Derby in breeding herds while the most prevalent phage types of the S. Typhimurium isolates were DT 12 and DT 120. The antimicrobial resistance analysis yielded a 35.2% of the isolates from the slaughter pigs resistant to one or more antimicrobials while 19.3% were...... resistant to four or more antimicrobials. A significantly higher percentage of resistance to antimicrobials was found in the S. Typhimurium isolates (χ2=4.72, p=0.029), where 42.9% presented resistance to one or more compounds. In breeding herds, just S. Typhimurium and S. 4,5],12:i: – isolates were tested......The objective of this paper is to analyse in further detail the Danish results of the EFSA baseline studies in slaughter pigs and breeding herds, and compare them with the results obtained in (1) the pre-implementation study that was carried out to establish the initial prevalence values...

Targeted deletion of the ara operon of Salmonella typhimurium enhances L-arabinose accumulation and drives PBAD-promoted expression of anti-cancer toxins and imaging agents.

Science.gov (United States)

Hong, Hyun; Lim, Daejin; Kim, Geun-Joong; Park, Seung-Hwan; Sik Kim, Hyeon; Hong, Yeongjin; Choy, Hyon E; Min, Jung-Joon

2014-01-01

Tumor-specific expression of antitumor drugs can be achieved using attenuated Salmonella typhimurium harboring the PBAD promoter, which is induced by L-arabinose. However, L-arabinose does not accumulate because it is metabolized to D-xylulose-5-P by enzymes encoded by the ara operon in Salmonellae. To address this problem, we developed an engineered strain of S. typhimurium in which the ara operon is deleted. Linear DNA transformation was performed using λ red recombinase to exchange the ara operon with linear DNA carrying an antibiotic-resistance gene with homology to regions adjacent to the ara operon. The ara operon-deleted strain and its parental strain were transformed with a plasmid encoding Renilla luciferase variant 8 (RLuc8) or cytolysin A (clyA) under the control of the PBAD promoter. Luciferase assays demonstrated that RLuc8 expression was 49-fold higher in the ara operon-deleted S. typhimurium than in the parental strain after the addition of L-arabinose. In vivo bioluminescence imaging showed that the tumor tissue targeted by the ara operon-deleted Salmonella had a stronger imaging signal (~30-fold) than that targeted by the parental strain. Mice with murine colon cancer (CT26) that had been injected with the ara operon-deleted S. typhimurium expressing clyA showed significant tumor suppression. The present report demonstrates that deletion of the ara operon of S. typhimurium enhances L-arabinose accumulation and thereby drives PBAD-promoted expression of cytotoxic agents and imaging agents. This is a promising approach for tumor therapy and imaging.

Effects of irradiation and fumaric acid treatment on the inactivation of Listeria monocytogenes and Salmonella typhimurium inoculated on sliced ham

Science.gov (United States)

Song, Hyeon-Jeong; Lee, Ji-Hye; Song, Kyung Bin

2011-11-01

To examine the effects of fumaric acid and electron beam irradiation on the inactivation of foodborne pathogens in ready-to-eat meat products, sliced ham was inoculated with Listeria monocytogenes and Salmonella typhimurium. The inoculated ham slices were treated with 0.5% fumaric acid or electron beam irradiation at 2 kGy. Fumaric acid treatment reduced the populations of L. monocytogenes and S. typhimurium by approximately 1 log CFU/g compared to control populations. In contrast, electron beam irradiation decreased the populations of S. typhimurium and L. monocytogenes by 3.78 and 2.42 log CFU/g, respectively. These results suggest that electron beam irradiation is a better and appropriate technique for improving the microbial safety of sliced ham.

Salmonella Typhimurium undergoes distinct genetic adaption during chronic infections of mice

DEFF Research Database (Denmark)

Søndberg, Emilie; Jelsbak, Lotte

2016-01-01

Background Typhoid fever caused by Salmonella enterica serovar Typhi (S. Typhi) is a severe systemic human disease and endemic in regions of the world with poor drinking water quality and sewage treatment facilities. A significant number of patients become asymptomatic life-long carriers of S....... In the current study genetic adaptation during experimental chronic S. Typhimurium infections of mice, an established model of chronic typhoid fever, was probed as an approach for studying the molecular mechanisms of host-adaptation during long-term host-association. Results Individually sequence-tagged wild...

ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR SCREENING OF MILK SAMPLES FOR SALMONELLA-TYPHIMURIUM IN DAIRY HERDS

DEFF Research Database (Denmark)

Hoorfar, Jeffrey; Wedderkopp, A.

1995-01-01

We investigated the ability of an antibody-specific, O antigen-based ELISA to document Salmonella typhimurium herd infections by screening of milk samples. Three cattle populations, 20 herds with no history of salmonellosis, 8 herds with history of S typhimurium epsiodes within the previous 7...... months, and 220 herds of unknown disease status, were tested. A herd was considered ELISA positive if at least 5% of the cows had OD values > 0.3. Among the 20 herds without history of salmonellosis, only 2 herds were ELISA positive, whereas all 8 herds with a known history of salmonellosis were ELISA...... positive (herd specificity, 0.9 and herd sensitivity, 1.0). A sig nificant correlation (P history of salmonellosis. It was concluded that ELISA testing of individual milk sam ples can be used for surveillance...

Isolation and identification of Salmonella spp. in drinking water, streams, and swine wastewater by molecular techniques in Taiwan

Science.gov (United States)

Kuo, C.; Hsu, B.; Shen, T.; Tseng, S.; Tsai, J.; Huang, K.; Kao, P.; Chen, J.

2013-12-01

Salmonella spp. is a common water-borne pathogens and its genus comprises more than 2,500 serotypes. Major pathogenic genotypes which cause typhoid fever, enteritis and other intestinal-type diseases are S. Typhimurium, S. Enteritidis, S. Stanley, S. Agona, S.Albany, S. Schwarzengrund, S. Newport, S. Choleraesuis, and S. Derby. Hence, the identification of the serotypes of Salmonella spp. is important. In the present study, the analytical procedures include direct concentration method, non-selective pre-enrichment method and selective enrichment method of Salmonella spp.. Both selective enrichment method and cultured bacteria were detected with specific primers of Salmonella spp. by polymerase chain reaction (PCR). At last, the serotypes of Salmonella were confirmed by using MLST (multilocus sequence typing) with aroC, dnaN, hemD, hisD, purE, sucA, thrA housekeeping genes to identify the strains of positive samples. This study contains 121 samples from three different types of water sources including the drinking water (51), streams (45), and swine wastewater (25). Thirteen samples with positive invA gene are separated from culture method. The strains of these positive samples which identified from MLST method are S. Albany, S. Typhimurium, S. Newport, S. Bareilly, and S. Derby. Some of the serotypes, S. Albany, S. Typhimurium and S. Newport, are highly pathogenic which correlated to human diarrhea. In our results, MLST is a useful method to identify the strains of Salmonella spp.. Keywords: Salmonella, PCR, MLST.

Impact of desiccation and heat exposure stress on Salmonella tolerance to acidic conditions.

Science.gov (United States)

Richardson, Kurt E; Cox, Nelson A; Cosby, Douglas E; Berrang, Mark E

2018-02-01

In a recent study, the pH of commonly used Salmonella pre-enrichment media became acidic (pH 4.0 to 5.0) when feed or feed ingredients were incubated for 24 h. Acidic conditions have been reported to injure or kill Salmonella. In this study, cultures of four known feed isolates (S. montevideo, S. senftenberg, S. tennessee, and S. schwarzengrund) and four important processing plant isolates (S. typhimurium, S. enteritidis, S. infantis, and S. heidelberg) were grown on meat and bone meal and later subjected to desiccation and heat exposure to stress the microorganism. The impact of stress on the isolates ability to survive in acidic conditions ranging from pH 4.0 to 7.0 was compared to the non-stressed isolate. Cell injury was determined on xylose lysine tergitol 4 (XLT4) and cell death determined on nutrient agar (NA). When measured by cell death in non-stressed Salmonella, S. typhimurium was the most acid tolerant and S. heidelberg was the most acid sensitive whereas in stressed Salmonella, S. senftenberg was the most acid tolerant and S. tennessee was the most acid sensitive. The pH required to cause cell injury varied among isolates. With some isolates, the pH required for 50% cell death and 50% cell injury was similar. In other isolates, cell injury occurred at a more neutral pH. These findings suggest that the pH of pre-enrichment media may influence the recovery and bias the serotype of Salmonella recovered from feed during pre-enrichment.

The two umuDC-like operons, samAB and umuDCST, in Salmonella typhimurium: The umuDCST operon may reduce UV-mutagenesis-promoting ability of the samAB operon

International Nuclear Information System (INIS)

Nohmi, Takehiko; Hakura, Atsushi; Watanabe, Masahiko; Yamada, Masami; Sofuni, Toshio; Nakai, Yasuharu; Murayama, Somay Y.

1993-01-01

Salmonella typhimurium, especially its derivatives containing pKM101 plasmid, has been widely used in the Ames test for the detection of environmental mutagens and carcinogens. It is known, however, that if the pKM101 plasmid is eliminated, S. typhimurium itself shows a much weaker mutagenic response to UV and some chemical mutagens than does Escherichia coli. In fact, certain potent base-change type mutagens, such as furylfuramide and aflatoxin B 1 , are nonmutagenic to S. typhimurium in the absence of pKM101, whereas they are strongly mutagenic to S. typhimurium in the presence of pKM101 plasmid as well as to E. coli. The low mutability can be restored to levels comparable to E. coli by introducing the plasmid carrying the E. coli umuDC operon or the pKM101 plasmid carrying mucAB operon. Salmonella typhimurium has an SOS regulatory system which resembles that of E. coli. Thus, it was suggested that S. typhimurium is deficient in the function of umuDC operon, which plays an essential role in UV and most chemical mutagenesis in E. coli. In order to clarify the implications of umuDC genes in mutagenesis and antimutagenesis in typhimurium, we have independently screened the umuDC-like genes of S. typhimurium TA1538. Consequently, we have cloned another umuDC-like operon which is 40% diverged from the aforementioned umuDC operon of S. typhimurium LT2 at the nucleotide level (16). We have termed the cloned DNA the samAB (Salmonella; mutagenesis) operon, and tentatively referred to the umuDC operon cloned from S. typhimurium LT2 (27,31) as the umuDC ST operon. Based on the results of the Southern hybridization experiment, we concluded that the two sets of umuDC-like operons reside in the same cells of S. typhimurium LT2 and TA1538. Our results also suggested that the umuDC ST operon reduces the UV-mutagenesis promoting ability of the samAB operon when the two operons are present on the same multi-copy number plasmid

Structure of the ribosomal interacting GTPase YjeQ from the enterobacterial species Salmonella typhimurium

International Nuclear Information System (INIS)

Nichols, C. E.; Johnson, C.; Lamb, H. K.; Lockyer, M.; Charles, I. G.; Hawkins, A. R.; Stammers, D. K.

2007-01-01

The X-ray crystal structure of the GTPase YjeQ from S. typhimurium is presented and compared with those of orthologues from T. maritima and B. subtilis. The YjeQ class of P-loop GTPases assist in ribosome biogenesis and also bind to the 30S subunit of mature ribosomes. YjeQ ribosomal binding is GTP-dependent and thought to specifically direct protein synthesis, although the nature of the upstream signal causing this event in vivo is as yet unknown. The attenuating effect of YjeQ mutants on bacterial growth in Escherichia coli makes it a potential target for novel antimicrobial agents. In order to further explore the structure and function of YjeQ, the isolation, crystallization and structure determination of YjeQ from the enterobacterial species Salmonella typhimurium (StYjeQ) is reported. Whilst the overall StYjeQ fold is similar to those of the previously reported Thematoga maritima and Bacillus subtilis orthologues, particularly the GTPase domain, there are larger differences in the three OB folds. Although the zinc-finger secondary structure is conserved, significant sequence differences alter the nature of the external surface in each case and may reflect varying signalling pathways. Therefore, it may be easier to develop YjeQ-specific inhibitors that target the N- and C-terminal regions, disrupting the metabolic connectivity rather than the GTPase activity. The availability of coordinates for StYjeQ will provide a significantly improved basis for threading Gram-negative orthologue sequences and in silico compound-screening studies, with the potential for the development of species-selective drugs

Structure of the ribosomal interacting GTPase YjeQ from the enterobacterial species Salmonella typhimurium

Energy Technology Data Exchange (ETDEWEB)

Nichols, C. E. [Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom); Johnson, C.; Lamb, H. K. [Institute of Cell and Molecular Biosciences, Catherine Cookson Building, Medical School, Framlington Place, Newcastle University, Newcastle-upon-Tyne NE2 4HH (United Kingdom); Lockyer, M. [Arrow Therapeutics Ltd, Britannia House, Trinity Street, Borough, London SE1 1DA (United Kingdom); Charles, I. G. [The Wolfson Institute for Biomedical Research, The Cruciform Building, University College London, Gower Street, London WC1E 6BT (United Kingdom); Hawkins, A. R. [Institute of Cell and Molecular Biosciences, Catherine Cookson Building, Medical School, Framlington Place, Newcastle University, Newcastle-upon-Tyne NE2 4HH (United Kingdom); Stammers, D. K., E-mail: daves@strubi.ox.ac.uk [Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN (United Kingdom)

2007-11-01

The X-ray crystal structure of the GTPase YjeQ from S. typhimurium is presented and compared with those of orthologues from T. maritima and B. subtilis. The YjeQ class of P-loop GTPases assist in ribosome biogenesis and also bind to the 30S subunit of mature ribosomes. YjeQ ribosomal binding is GTP-dependent and thought to specifically direct protein synthesis, although the nature of the upstream signal causing this event in vivo is as yet unknown. The attenuating effect of YjeQ mutants on bacterial growth in Escherichia coli makes it a potential target for novel antimicrobial agents. In order to further explore the structure and function of YjeQ, the isolation, crystallization and structure determination of YjeQ from the enterobacterial species Salmonella typhimurium (StYjeQ) is reported. Whilst the overall StYjeQ fold is similar to those of the previously reported Thematoga maritima and Bacillus subtilis orthologues, particularly the GTPase domain, there are larger differences in the three OB folds. Although the zinc-finger secondary structure is conserved, significant sequence differences alter the nature of the external surface in each case and may reflect varying signalling pathways. Therefore, it may be easier to develop YjeQ-specific inhibitors that target the N- and C-terminal regions, disrupting the metabolic connectivity rather than the GTPase activity. The availability of coordinates for StYjeQ will provide a significantly improved basis for threading Gram-negative orthologue sequences and in silico compound-screening studies, with the potential for the development of species-selective drugs.

Salmonella enterica Serovar Typhimurium in Mauritius Linked to Consumption of Marlin Mousse

DEFF Research Database (Denmark)

Issack, M. I.; Hendriksen, Rene S.; Lun, P. L. K.

2009-01-01

. A laboratory investigation of one sample of marlin mousse manufactured 3 days later, and the individual ingredients sampled a week after production of the contaminated batch were all negative for Salmonella. Serotyping and minimum inhibitory concentration determination were performed on 12 patient isolates......We report the first outbreak of salmonellosis caused by consumption of contaminated marlin mousse. Between 29 October and 5 November 2008, at least 53 persons developed diarrheal illness, all with a history of eating marlin mousse. Salmonella spp. that did not produce gas from glucose was isolated...... from stools of 26 affected patients and blood culture from one patient. Salmonella sp. isolates with the same phenotype were isolated in three samples of marlin mousse manufactured on 27 October 2008. The constituents of the mousse were smoked marlin, raw eggs, bovine gelatin, oil, and cream...

Consumer attitudes and behaviours--key risk factors in an outbreak of Salmonella typhimurium phage type 12 infection sourced to chicken nuggets.

Science.gov (United States)

Kenny, B; Hall, R; Cameron, S

1999-04-01

To identify the source and intervention methods for an outbreak of Salmonella Typhimurium phage type 12 in South Australia. Ten cases of S. Typhimurium phage type (PT) 12 infection were notified in South Australia in a four-week period from 7 May 1998. Nine cases and 27 controls were included in a case control study to test the hypothesis that illness was associated with the consumption of chicken nuggets. A significant association between illness and the consumption of one brand of chicken nuggets was determined, odds ratio undefined (95% CI undefined; p = undefined). Nine of nine cases and one of 27 controls reported eating these chicken nuggets. S. Typhimurium PT 12 was isolated from an opened sample of this particular brand of nuggets which had been retrieved from the home of one case. The implicated nuggets were essentially a raw product which had been 'flash fried' in contrast with other brands which were fully cooked. The investigation highlighted issues of inadequate labelling and consumer responses to labelling information which affect food safety. A media release to highlight to the consumer the need to cook frozen food properly and a voluntary recall of the 'flash fried' product was instigated as a result of these conclusions. Further action is needed to eliminate the potential hazard that consumers will perceive and handle 'flash fried' nuggets as if they are a cooked chicken product.

Diversity of Salmonella isolates from central Florida surface waters.

Science.gov (United States)

McEgan, Rachel; Chandler, Jeffrey C; Goodridge, Lawrence D; Danyluk, Michelle D

2014-11-01

Identification of Salmonella serotypes is important for understanding the environmental diversity of the genus Salmonella. This study evaluates the diversity of Salmonella isolates recovered from 165 of 202 Central Florida surface water samples and investigates whether the serotype of the environmental Salmonella isolates can be predicted by a previously published multiplex PCR assay (S. Kim, J. G. Frye, J. Hu, P. J. Fedorka-Cray, R. Gautom, and D. S. Boyle, J. Clin. Microbiol. 44:3608-3615, 2006, http://dx.doi.org/10.1128/JCM.00701-06). Multiplex PCR was performed on 562 Salmonella isolates (as many as 36 isolates per water sample) to predict serotypes. Kauffmann-White serogrouping was used to confirm multiplex PCR pattern groupings before isolates were serotyped, analyzed by pulsed-field gel electrophoresis, and assayed for antimicrobial susceptibility. In 41.2% of the Salmonella-positive water samples, all Salmonella isolates had identical multiplex PCR patterns; in the remaining 58.8%, two or more multiplex PCR patterns were identified. Within each sample, isolates with matching multiplex PCR patterns had matching serogroups. The multiplex patterns of 495 isolates (88.1%) did not match any previously reported pattern. The remaining 68 isolates matched reported patterns but did not match the serotypes for those patterns. The use of the multiplex PCR allowed the number of isolates requiring further analysis to be reduced to 223. Thirty-three Salmonella enterica serotypes were identified; the most frequent included serotypes Muenchen, Rubislaw, Anatum, Gaminara, and IV_50:z4,z23:-. A majority (141/223) of Salmonella isolates clustered into one genotypic group. Salmonella isolates in Central Florida surface waters are serotypically, genotypically, and phenotypically (in terms of antimicrobial susceptibility) diverse. While isolates could be grouped as different or potentially the same using multiplex PCR, the multiplex PCR pattern did not predict the Salmonella

Isolation of Salmonella typhimurium strains that utilize exogenous 3-deoxy-D-manno-Octulosonate for synthesis of lipopolysaccharide

International Nuclear Information System (INIS)

Goldman, R.C.; Devine, E.M.

1987-01-01

Spontaneous mutants of Salmonella typhimurium LT2 were selected for the ability to accumulate exogenous 3-deoxy-D-manno-octulosonate (KDO). Bacteria containing a gene (kdsA) which codes for a temperature-sensitive KDO-8-phosphate synthetase were plated at the restrictive temperature of 42 0 C on medium containing 5 mM KDO. Since bacteria containing the kdsA lesion are unable to grown at 42 0 C due to inhibition of lipopolysaccharide (LPS) synthesis and accumulation of lipid A precursor, this method allowed direct, positive selection of mutants capable of utilizing exogenous KDO for LPS synthesis. Spontaneous mutants, selected at a frequency of about 10 -6 , required exogenous KDO for growth at 42 0 C. The growth rate at 42 0 C was nearly normal in the presence of 20 mM KDO and was directly proportional to KDO concentrations below 20 mM. Exogenous KDO also suppressed accumulation of lipid A precursor. The apparent K/sub m/ for KDO accumulation was 23 mM, and the maximum rate of transport was calculated to be 505 pmol of DKO per min per 10 8 cells. Bacteria incorporated exogenous [ 3 H]KDO exclusively into LPS, with less that 10% dilution in specific activity due to residual endogenous KDO synthesis. The mutation giving rise to the ability to accumulate exogenous KDO was extremely useful in the direct screening for new mutations in the kdsA gene after localized mutagenesis. Five mutations in kdsA were isolated, four of which were new alleles as determined by on fine-structure analysis. The ability to introduce labeled ( 3 H, 13 C, and 14 C) KDO in vivo should simplify and extend the analysis of this critical metabolic pathway in gram-negative bacteria

Modification of Salmonella Typhimurium motility by the probiotic yeast strain Saccharomyces boulardii.

Directory of Open Access Journals (Sweden)

Rodolphe Pontier-Bres

Full Text Available BACKGROUND: Motility is an important component of Salmonella enterica serovar Typhimurium (ST pathogenesis allowing the bacteria to move into appropriate niches, across the mucus layer and invade the intestinal epithelium. In vitro, flagellum-associated motility is closely related to the invasive properties of ST. The probiotic yeast Saccharomyces boulardii BIOCODEX (S.b-B is widely prescribed for the prophylaxis and treatment of diarrheal diseases caused by bacteria or antibiotics. In case of Salmonella infection, S.b-B has been shown to decrease ST invasion of T84 colon cell line. The present study was designed to investigate the impact of S.b-B on ST motility. METHODOLOGY/PRINCIPAL FINDINGS: Experiments were performed on human colonic T84 cells infected by the Salmonella strain 1344 alone or in the presence of S.b-B. The motility of Salmonella was recorded by time-lapse video microscopy. Next, a manual tracking was performed to analyze bacteria dynamics (MTrackJ plugin, NIH image J software. This revealed that the speed of bacterial movement was modified in the presence of S.b-B. The median curvilinear velocity (CLV of Salmonella incubated alone with T84 decreased from 43.3 µm/sec to 31.2 µm/sec in the presence of S.b-B. Measurement of track linearity (TL showed similar trends: S.b-B decreased by 15% the number of bacteria with linear tract (LT and increased by 22% the number of bacteria with rotator tract (RT. Correlation between ST motility and invasion was further established by studying a non-motile flagella-deficient ST strain. Indeed this strain that moved with a CLV of 0.5 µm/sec, presented a majority of RT and a significant decrease in invasion properties. Importantly, we show that S.b-B modified the motility of the pathogenic strain SL1344 and significantly decreased invasion of T84 cells by this strain. CONCLUSIONS: This study reveals that S.b-B modifies Salmonella's motility and trajectory which may account for the modification

Modification of Salmonella Typhimurium Motility by the Probiotic Yeast Strain Saccharomyces boulardii

Science.gov (United States)

Pontier-Bres, Rodolphe; Prodon, François; Munro, Patrick; Rampal, Patrick; Lemichez, Emmanuel; Peyron, Jean François; Czerucka, Dorota

2012-01-01

Background Motility is an important component of Salmonella enterica serovar Typhimurium (ST) pathogenesis allowing the bacteria to move into appropriate niches, across the mucus layer and invade the intestinal epithelium. In vitro, flagellum-associated motility is closely related to the invasive properties of ST. The probiotic yeast Saccharomyces boulardii BIOCODEX (S.b-B) is widely prescribed for the prophylaxis and treatment of diarrheal diseases caused by bacteria or antibiotics. In case of Salmonella infection, S.b-B has been shown to decrease ST invasion of T84 colon cell line. The present study was designed to investigate the impact of S.b-B on ST motility. Methodology/Principal Findings Experiments were performed on human colonic T84 cells infected by the Salmonella strain 1344 alone or in the presence of S.b-B. The motility of Salmonella was recorded by time-lapse video microscopy. Next, a manual tracking was performed to analyze bacteria dynamics (MTrackJ plugin, NIH image J software). This revealed that the speed of bacterial movement was modified in the presence of S.b-B. The median curvilinear velocity (CLV) of Salmonella incubated alone with T84 decreased from 43.3 µm/sec to 31.2 µm/sec in the presence of S.b-B. Measurement of track linearity (TL) showed similar trends: S.b-B decreased by 15% the number of bacteria with linear tract (LT) and increased by 22% the number of bacteria with rotator tract (RT). Correlation between ST motility and invasion was further established by studying a non-motile flagella-deficient ST strain. Indeed this strain that moved with a CLV of 0.5 µm/sec, presented a majority of RT and a significant decrease in invasion properties. Importantly, we show that S.b-B modified the motility of the pathogenic strain SL1344 and significantly decreased invasion of T84 cells by this strain. Conclusions This study reveals that S.b-B modifies Salmonella's motility and trajectory which may account for the modification of Salmonella

A community effort towards a knowledge-base and mathematical model of the human pathogen Salmonella Typhimurium LT2

Directory of Open Access Journals (Sweden)

Shin Sook-Il

2011-01-01

Full Text Available Abstract Background Metabolic reconstructions (MRs are common denominators in systems biology and represent biochemical, genetic, and genomic (BiGG knowledge-bases for target organisms by capturing currently available information in a consistent, structured manner. Salmonella enterica subspecies I serovar Typhimurium is a human pathogen, causes various diseases and its increasing antibiotic resistance poses a public health problem. Results Here, we describe a community-driven effort, in which more than 20 experts in S. Typhimurium biology and systems biology collaborated to reconcile and expand the S. Typhimurium BiGG knowledge-base. The consensus MR was obtained starting from two independently developed MRs for S. Typhimurium. Key results of this reconstruction jamboree include i development and implementation of a community-based workflow for MR annotation and reconciliation; ii incorporation of thermodynamic information; and iii use of the consensus MR to identify potential multi-target drug therapy approaches. Conclusion Taken together, with the growing number of parallel MRs a structured, community-driven approach will be necessary to maximize quality while increasing adoption of MRs in experimental design and interpretation.

A community effort towards a knowledge-base and mathematical model of the human pathogen Salmonella Typhimurium LT2.

Science.gov (United States)

Thiele, Ines; Hyduke, Daniel R; Steeb, Benjamin; Fankam, Guy; Allen, Douglas K; Bazzani, Susanna; Charusanti, Pep; Chen, Feng-Chi; Fleming, Ronan M T; Hsiung, Chao A; De Keersmaecker, Sigrid C J; Liao, Yu-Chieh; Marchal, Kathleen; Mo, Monica L; Özdemir, Emre; Raghunathan, Anu; Reed, Jennifer L; Shin, Sook-il; Sigurbjörnsdóttir, Sara; Steinmann, Jonas; Sudarsan, Suresh; Swainston, Neil; Thijs, Inge M; Zengler, Karsten; Palsson, Bernhard O; Adkins, Joshua N; Bumann, Dirk

2011-01-18

Metabolic reconstructions (MRs) are common denominators in systems biology and represent biochemical, genetic, and genomic (BiGG) knowledge-bases for target organisms by capturing currently available information in a consistent, structured manner. Salmonella enterica subspecies I serovar Typhimurium is a human pathogen, causes various diseases and its increasing antibiotic resistance poses a public health problem. Here, we describe a community-driven effort, in which more than 20 experts in S. Typhimurium biology and systems biology collaborated to reconcile and expand the S. Typhimurium BiGG knowledge-base. The consensus MR was obtained starting from two independently developed MRs for S. Typhimurium. Key results of this reconstruction jamboree include i) development and implementation of a community-based workflow for MR annotation and reconciliation; ii) incorporation of thermodynamic information; and iii) use of the consensus MR to identify potential multi-target drug therapy approaches. Taken together, with the growing number of parallel MRs a structured, community-driven approach will be necessary to maximize quality while increasing adoption of MRs in experimental design and interpretation.

A community effort towards a knowledge-base and mathematical model of the human pathogen Salmonella Typhimurium LT2

Energy Technology Data Exchange (ETDEWEB)

Thiele, Ines; Hyduke, Daniel R.; Steeb, Benjamin; Fankam, Guy; Allen, Douglas K.; Bazzani, Susanna; Charusanti, Pep; Chen, Feng-Chi; Fleming, Ronan MT; Hsiung, Chao A.; De Keersmaecker, Sigrid CJ; Liao, Yu-Chieh; Marchal, Kathleen; Mo, Monica L.; Özdemir, Emre; Raghunathan, Anu; Reed, Jennifer L.; Shin, Sook-Il; Sigurbjörnsdóttir, Sara; Steinmann, Jonas; Sudarsan, Suresh; Swainston, Neil; Thijs, Inge M.; Zengler, Karsten; Palsson, Bernhard O.; Adkins, Joshua N.; Bumann, Dirk

2011-01-01

Metabolic reconstructions (MRs) are common denominators in systems biology and represent biochemical, genetic, and genomic (BiGG) knowledge-bases for target organisms by capturing currently available information in a consistent, structured manner. Salmonella enterica subspecies I serovar Typhimurium is a human pathogen, causes various diseases and its increasing antibiotic resistance poses a public health problem. Here, we describe a community-driven effort, in which more than 20 experts in S. Typhimurium biology and systems biology collaborated to reconcile and expand the S. Typhimurium BiGG knowledge-base. The consensus MR was obtained starting from two independently developed MRs for S. Typhimurium. Key results of this reconstruction jamboree include i) development and implementation of a community-based workflow for MR annotation and reconciliation; ii) incorporation of thermodynamic information; and iii) use of the consensus MR to identify potential multi-target drug therapy approaches. Finally, taken together, with the growing number of parallel MRs a structured, community-driven approach will be necessary to maximize quality while increasing adoption of MRs in experimental design and interpretation.

Effect of gamma irradiation on chemical and biological properties of lipopolysaccharide from Salmonella typhimurium

International Nuclear Information System (INIS)

Naidu, Mamta D.; Chander, Ramesh; Nair, P.M.

1998-01-01

Lipopolysaccharide (LPS) from S. typhimurium on exposure to γ-radiation resulted in decrease in toxicity and was less mitogenic. Silver stained profiles of irradiated LPS on polyacrylamide gels revealed complete loss of its heteropolysaccharides which was confirmed further by analysing lipid A and LPS from Salmonella minnesota Re mutants on SDS-PAGE. Glucosamine and 2-keto 3-deoxy-octonate (Kdo) contents were significantly decreased on treatment. Lipid A obtained by removal of heteropolysaccharides from LPS was less toxic on exposure to gamma radiations. (author)

The influence of glycerol on γ-induced mutagenesis in Salmonella typhimurium cells

International Nuclear Information System (INIS)

Basha, S.G.; Krasavin, E.A.; Kozubek, S.; Amirtaev, K.G.

1990-01-01

A study was made of the modifying effect of glycerol on the survival rate and γ-radiation-induced mutagenesis of Salmonella typhimurium cells TA98, TA100 and TA102. The DMF value, with respect to the survival rate, was 2.05-0.20. The dependence of the yield of γ-radiation-induced mutants on radiation dose was described by the curve with a maximum; the mutation frequency M(D) was well described by a gradual function M(D)=kD x . DMF values of the induced mutagenesis amounted to 2 for strains TA100 and TA102, and 1.5 for strain TA98

Complete Genome Sequence of a Human-Invasive Salmonella enterica Serovar Typhimurium Strain of the Emerging Sequence Type 213 Harboring a Multidrug Resistance IncA/C Plasmid and a blaCMY-2-Carrying IncF Plasmid.

Science.gov (United States)

Silva, Claudia; Calva, Edmundo; Calva, Juan J; Wiesner, Magdalena; Fernández-Mora, Marcos; Puente, José L; Vinuesa, Pablo

2015-11-12

Salmonella enterica subsp. enterica serovar Typhimurium strain 33676 was isolated in Mexico City, Mexico, from a patient with a systemic infection, and its complete genome sequence was determined using PacBio single-molecule real-time technology. Strain 33676 harbors an IncF plasmid carrying the extended-spectrum cephalosporin gene blaCMY-2 and a multidrug resistance IncA/C plasmid. Copyright © 2015 Silva et al.

Construction of genetic markers for the study of Salmonella typhimurium infection of murine macrophages

DEFF Research Database (Denmark)

Jelsbak, Lotte; Olsen, John Elmerdahl

in combination with available host markers it will be possible to estimate the time-point at which a specific gene is required for progression of SCV maturation. These developmentally regulated reporter fusions constitute a set of novel developmental markers for the study of Salmonella Typhimurium infection...... with the host cell, (2) Formation of early SCV, (3) Maturation into late SCV, (4) Initiation of bacterial replication, (5) Formation of Sifs. In this project, we have constructed a set of reporter fusions which are temporally and spatially regulated during the progression of SCV maturation. The reporter fusions...... were constructed using Red-mediated recombination (1) and the promoters were selected from the recently published expressional data of Salmonella infection of murine macrophages (2). As reporter proteins we both use a stable GFPmut3 variant as well as an unstable GFP variant (3). Using these fusions...

In Vitro Development of Ciprofloxacin Resistance of Salmonella enterica Serovars Typhimurium, Enteritidis, and Indiana Isolates from Food Animals.

Science.gov (United States)

Zhang, Wen-Hui; Zhang, Chuan-Zhen; Liu, Zhi-Jie; Gu, Xi-Xi; Li, Wan; Yang, Ling; Liu, Ya-Hong; Zeng, Zhen-Ling; Jiang, Hong-Xia

2017-09-01

Difference in the development of resistance may be associated with the epidemiological spread and drug resistance of different Salmonella enterica serovar strains. In the present study, three susceptible S. enterica serovars, Typhimurium (ST), Enteritidis (SE), and Indiana (SI) strains, were subjected to stepwise selection with increasing ciprofloxacin concentrations. The results indicated that the mutation frequencies of the SI group were 10 1 -10 4 higher and developed resistance to ciprofloxacin more rapidly compared with the ST and SE groups. Ciprofloxacin accumulation in the SI strain was also higher than the other two strains in the presence of an efflux pump inhibitor. The development of ciprofloxacin resistance was quite different among the three serovar strains. In SI, increasing AcrAB-TolC efflux pump expression and single or double mutations in gyrA with or without a single parC mutation (T57S) were found in the development of ciprofloxacin resistance. In SE, an increase in the AcrAB-TolC efflux pump regulatory gene ramA gradually decreased as resistant bacteria developed; then resistance resulted from gyrA D87G and gyrB E466D mutations and/or in other active efflux pumps besides AcrAB-TolC. For ST, ramA expression increased rapidly along with gyrA D87 N and/or gyrB S464F mutations. In conclusion, persistent use of ciprofloxacin may aggravate the resistance of different S. enterica serovars and prudent use of the fluoroquinolones is needed. The quicker resistance and higher mutation frequency of the SI isolates present a potential public health threat.

Comparative immunological evaluation of recombinant Salmonella Typhimurium strains expressing model antigens as live oral vaccines.

Science.gov (United States)

Zheng, Song-yue; Yu, Bin; Zhang, Ke; Chen, Min; Hua, Yan-Hong; Yuan, Shuofeng; Watt, Rory M; Zheng, Bo-Jian; Yuen, Kwok-Yung; Huang, Jian-Dong

2012-09-26

Despite the development of various systems to generate live recombinant Salmonella Typhimurium vaccine strains, little work has been performed to systematically evaluate and compare their relative immunogenicity. Such information would provide invaluable guidance for the future rational design of live recombinant Salmonella oral vaccines. To compare vaccine strains encoded with different antigen delivery and expression strategies, a series of recombinant Salmonella Typhimurium strains were constructed that expressed either the enhanced green fluorescent protein (EGFP) or a fragment of the hemagglutinin (HA) protein from the H5N1 influenza virus, as model antigens. The antigens were expressed from the chromosome, from high or low-copy plasmids, or encoded on a eukaryotic expression plasmid. Antigens were targeted for expression in either the cytoplasm or the outer membrane. Combinations of strategies were employed to evaluate the efficacy of combined delivery/expression approaches. After investigating in vitro and in vivo antigen expression, growth and infection abilities; the immunogenicity of the constructed recombinant Salmonella strains was evaluated in mice. Using the soluble model antigen EGFP, our results indicated that vaccine strains with high and stable antigen expression exhibited high B cell responses, whilst eukaryotic expression or colonization with good construct stability was critical for T cell responses. For the insoluble model antigen HA, an outer membrane expression strategy induced better B cell and T cell responses than a cytoplasmic strategy. Most notably, the combination of two different expression strategies did not increase the immune response elicited. Through systematically evaluating and comparing the immunogenicity of the constructed recombinant Salmonella strains in mice, we identified their respective advantages and deleterious or synergistic effects. Different construction strategies were optimally-required for soluble versus

Salmonella Typhimurium ST213 is associated with two types of IncA/C plasmids carrying multiple resistance determinants.

Science.gov (United States)

Wiesner, Magdalena; Calva, Edmundo; Fernández-Mora, Marcos; Cevallos, Miguel A; Campos, Freddy; Zaidi, Mussaret B; Silva, Claudia

2011-01-11

Salmonella Typhimurium ST213 was first detected in the Mexican Typhimurium population in 2001. It is associated with a multi-drug resistance phenotype and a plasmid-borne blaCMY-2 gene conferring resistance to extended-spectrum cephalosporins. The objective of the current study was to examine the association between the ST213 genotype and blaCMY-2 plasmids. The blaCMY-2 gene was carried by an IncA/C plasmid. ST213 strains lacking the blaCMY-2 gene carried a different IncA/C plasmid. PCR analysis of seven DNA regions distributed throughout the plasmids showed that these IncA/C plasmids were related, but the presence and absence of DNA stretches produced two divergent types I and II. A class 1 integron (dfrA12, orfF and aadA2) was detected in most of the type I plasmids. Type I contained all the plasmids carrying the blaCMY-2 gene and a subset of plasmids lacking blaCMY-2. Type II included all of the remaining blaCMY-2-negative plasmids. A sequence comparison of the seven DNA regions showed that both types were closely related to IncA/C plasmids found in Escherichia, Salmonella, Yersinia, Photobacterium, Vibrio and Aeromonas. Analysis of our Typhimurium strains showed that the region containing the blaCMY-2 gene is inserted between traA and traC as a single copy, like in the E. coli plasmid pAR060302. The floR allele was identical to that of Newport pSN254, suggesting a mosaic pattern of ancestry with plasmids from other Salmonella serovars and E. coli. Only one of the tested strains was able to conjugate the IncA/C plasmid at very low frequencies (10-7 to 10-9). The lack of conjugation ability of our IncA/C plasmids agrees with the clonal dissemination trend suggested by the chromosomal backgrounds and plasmid pattern associations. The ecological success of the newly emerging Typhimurium ST213 genotype in Mexico may be related to the carriage of IncA/C plasmids. We conclude that types I and II of IncA/C plasmids originated from a common ancestor and that the

Global monitoring of Salmonella serovar distribution from the World Health Organization Global Foodborne Infections Network Country Data Bank: results of quality assured laboratories from 2001 to 2007.

Science.gov (United States)

Hendriksen, Rene S; Vieira, Antonio R; Karlsmose, Susanne; Lo Fo Wong, Danilo M A; Jensen, Arne B; Wegener, Henrik C; Aarestrup, Frank M

2011-08-01

Salmonella enterica is commonly acquired from contaminated food and is an important cause of illness worldwide. Interventions are needed to control Salmonella; subtyping Salmonella by serotyping is useful for targeting such interventions. We, therefore, analyzed the global distribution of the 15 most frequently identified serovars of Salmonella isolated from humans from 2001 to 2007 in laboratories from 37 countries that participated in World Health Organization Global Foodborne Infections Network and demonstrated serotyping proficiency in the Global Foodborne Infections Network External Quality Assurance System. In all regions throughout the study period, with the exception of the Oceania and North American regions, Salmonella serovars Enteritidis and Typhimurium ranked as the most common and second most common serovar, respectively. In the North American and Oceania (Australia and New Zealand) regions, Salmonella serovar Typhimurium was the most common serovar reported, and Salmonella serovar Enteritidis was the second most common serovar. During the study period, the proportion of Salmonella isolates reported from humans that were Salmonella serovar Enteritidis was 43.5% (range: 40.6% [2007] to 44.9% [2003]), and Salmonella serovar Typhimurium was 17.1% (range: 15% [2007] to 18.9% [2001]). Salmonella serovars Newport (mainly observed in Latin and North American and European countries), Infantis (dominating in all regions), Virchow (mainly observed in Asian, European, and Oceanic countries), Hadar (profound in European countries), and Agona (intense in Latin and North American and European countries) were also frequently isolated with an overall proportion of 3.5%, 1.8%, 1.5%, 1.5%, and 0.8%, respectively. There were large differences in the most commonly isolated serovars between regions, but lesser differences between countries within the same region. The results also highlight the complexity of the global epidemiology of Salmonella and the need and importance

Role of subtyping in detecting Salmonella cross contamination in the laboratory.

LENUS (Irish Health Repository)

De Lappe, Niall

2009-01-01

BACKGROUND: With the exception of M. tuberculosis, little has been published on the problems of cross-contamination in bacteriology laboratories. We performed a retrospective analysis of subtyping data from the National Salmonella Reference Laboratory (Ireland) from 2000-2007 to identify likely incidents of laboratory cross contamination. METHODS: Serotyping and antimicrobial susceptibility testing was performed on all Salmonella isolates received in the NSRL. Phage typing was performed on all S. Typhimurium and S. Enteritidis isolates while multi-locus variance analysis (MLVA) was performed on selected S. Typhimurium isolates. Pulsed field gel electrophoresis (PFGE) using the PulseNet standard protocol was performed on selected isolates of various serovars. RESULTS: Twenty-three incidents involving fifty-six isolates were identified as likely to represent cross contamination. The probable sources of contamination identified were the laboratory positive control isolate (n = 13), other test isolates (n = 9) or proficiency test samples (n = 1). CONCLUSION: The scale of laboratory cross-contamination in bacteriology is most likely under recognized. Testing laboratories should be aware of the potential for cross-contamination, regularly review protocols to minimize its occurrence and consider it as a possibility when unexpected results are obtained.

The type VI secretion system encoded in SPI-6 plays a role in gastrointestinal colonization and systemic spread of Salmonella enterica serovar Typhimurium in the chicken.

Directory of Open Access Journals (Sweden)

David Pezoa

Full Text Available The role of the Salmonella Pathogenicity Islands (SPIs in pathogenesis of Salmonella enterica Typhimurium infection in the chicken is poorly studied, while many studies have been completed in murine models. The Type VI Secretion System (T6SS is a recently described protein secretion system in Gram-negative bacteria. The genus Salmonella contains five phylogenetically distinct T6SS encoded in differentially distributed genomic islands. S. Typhimurium harbors a T6SS encoded in SPI-6 (T6SSSPI-6, which contributes to the ability of Salmonella to colonize mice. On the other hand, serotype Gallinarum harbors a T6SS encoded in SPI-19 (T6SSSPI-19 that is required for colonization of chicks. In this work, we investigated the role of T6SSSPI-6 in infection of chicks by S. Typhimurium. Oral infection of White Leghorn chicks showed that a ΔT6SSSPI-6 mutant had reduced colonization of the gut and internal organs, compared with the wild-type strain. Transfer of the intact T6SSSPI-6 gene cluster into the T6SS mutant restored bacterial colonization. In addition, our results showed that transfer of T6SSSPI-19 from S. Gallinarum to the ΔT6SSSPI-6 mutant of S. Typhimurium not only complemented the colonization defect but also resulted in a transient increase in the colonization of the cecum and ileum of chicks at days 1 and 3 post-infection. Our data indicates that T6SSSPI-6 contributes to chicken colonization and suggests that both T6SSSPI-6 and T6SSSPI-19 perform similar functions in vivo despite belonging to different phylogenetic families.

Molecular epidemiology and antimicrobial resistance of Salmonella Typhimurium DT104 on Ontario swine farms

Science.gov (United States)

Farzan, Abdolvahab; Friendship, Robert M.; Poppe, Cornelis; Martin, Laura; Dewey, Catherine E.; Funk, Julie

2008-01-01

This study was conducted to examine antimicrobial resistances, plasmid profiles, and pulsed-field gel electrophoresis patterns of 80 Salmonella Typhimurium (including var. Copenhagen) DT104 strains (including DT104a and DT104b) recovered from pig and environmental fecal samples on 17 swine farms in Ontario. No resistance was observed to amoxicillin/clavulanic acid, apramycin, carbadox, cephalothin, ceftriaxone, ceftiofur, cefoxitin, ciprofloxacin, nalidixic acid, trimethoprim, and tobramycin. However, the isolates exhibited resistance against 4 to 10 antimicrobials with the most frequent resistance being to sulfonamides (Su), ampicillin (A), streptomycin (S), spectinomycin (Sp), chloramphenicol (C), tetracycline (T), and florfenicol (F). Thirteen distinct resistance patterns were determined but 88% of isolates shared the typical resistance pattern “ACSpSSuT.” Twelve different plasmid profiles were observed; the 62 MDa virulence-associated plasmid was detected in 95% of the isolates. The 2.1 MDa plasmid was the second most frequent one, which was harbored by 65% isolates. The isolates were classified into 23 distinct genotypes by PFGE-SpeI + BlnI when difference in at least one fragment was defined as a distinct genotype. In total, 39 distinct “types” were observed when defining a “type” based on the combination of antimicrobial resistance, plasmid pattern, and PFGE-SpeI + BlnI for each isolate. The highest diversity was 0.96 (95% CI: 0.92, 0.96) for the “type” described above followed by 0.92 (95% CI: 0.88, 0.93) for PFGE-SpeI + BlnI. The diversity of DT104 isolates indicates there might be multiple sources for this microorganism on swine farms. This knowledge might be used to track these sources, as well as to study the extent of human salmonellosis attributed to pork compared to food products derived from other food-producing animals. PMID:18505209

Effects of gamma radiation and heat on immunogenicity and morphology of Salmonella typhimurium

International Nuclear Information System (INIS)

Santos Araujo, E. dos; Silva, D.M.

1979-01-01

Results are presented about immunogenicity of vaccines prepared from Salmonella typhimurium suspensions submitted to 0,55 MR and 1,10 MR of gamma radiation and heating at 60 0 C, (60 min) and at 100 0 C, (3 min) correlated with the cell morphology alterations observed at electron microscopy. The results of mouse-protection tests showed that prepared vaccines with 0,55 MR and with heating at 60 0 C, 60 min were identical and more efficient than the two other treatments. The electronmicrografies also showed one positive correlation between morphological alterations caused by the treatments and the immunogenicities. (Author) [pt

Effects of P22 bacteriophage on salmonella Enterica subsp. enterica serovar Typhimurium DMC4 strain biofilm formation and eradication

Directory of Open Access Journals (Sweden)

Karaca Basar

2015-01-01

Full Text Available Over the last decades, several antimicrobial agents have been made available. Due to increasing antimicrobial resistance, bacteriophages were rediscovered for their potential applications against bacterial infections. In the present study, biofilm inhibition and eradication of Salmonella enterica subsp. enterica serovar Typhimurium DMC4 strain (S. Typhimurium was evaluated with respect to different incubation periods at different P22 phage titrations. The efficacy of P22 phage on biofilm formation and eradication of S. Typhimurium DMC4 strain was screened in vitro on polystyrene and stainless steel surfaces. The biofilm forming capacity of S. Typhimurium was significantly reduced at higher phage titrations (106 pfu/mL ≤. All phage titers (104-108 pfu/mL were found to be effective at the end of the 24 h-incubation period whereas higher phage titrations were found to be effective at the end of the 48 h and 72 h of incubation. P22 phage has less efficacy on already formed, especially mature biofilms (72 h-old biofilm. Notable results of P22 phage treatment on S. Typhimurium biofilm suggest that P22 phage has potential uses in food systems.

Effect of Salvia chorassanica Root Aqueous, Ethanolic and Hydro Alcoholic Extracts on Staphylococcus aureus, Enterococcus faecalis, Salmonella typhimurium and Escherichia coli

Directory of Open Access Journals (Sweden)

Azam Mehraban

2016-11-01

Full Text Available Background Nowadays, through the previous researches, it has become clear that Salvia has important health benefits. Salvia chorassanica is one of the valuable native Iranian species which only grows in Khorasan province, Iran. Objectives The aim of this study is to evaluate the antimicrobial effect of Salvia chorassanica root aqueous, ethanolic and hydro alcoholic extracts on Staphylococcus aureus, Enterococcus faecalis, Salmonella typhimurium and Escherichia coli. Methods In this experimental study, maceration method was used to prepare extracts. Study setup was conducted in March 2014.The duration of study setup took for two months. The micro dilution method by ELISA was used to determine the minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC of aqueous, ethanolic and hydro alcoholic extracts of root of Salvia chorassanica against Staphylococcus aureus, Enterococcus faecalis, Salmonella typhimurium and Escherichia coli. The antibacterial effect also was evaluated using agar diffusion method. The inhibition zones of growth against the extracts were measured in comparison to standards antibiotics. Chloramphenicol as positive control on Enterococcus faecalis, Tetracycline on Staphylococcus aureus, Gentamicin on Escherichia coli and Neomycin on Salmonella typhimurium. The data were analyzed using one way analysis of variance (ANOVA with SPSS version 16. Results The highest inhibition zone in diffusion method was related to ethanolic extract of Salvia chorassanica root against Gram-positive bacteria, Staphylococcus aureus and Enterococcus faecalis. The calculated MIC in aqueous and ethanolic extracts of root for Staphylococcus aureus was 240 and 120 mg/mL, for Enterococcus faecalis was 120 and 60 mg/mL respectively, and for Escherichia coli and Salmonella typhimurium was equal to 240 mg/mL. The amount in hydro alcoholic extracts for Gram-positive bacteria was 60 mg/mL and for Gram-negative bacteria was 120 mg/mL. The

Aptasensors for rapid detection of Escherichia coli O157:H7 and Salmonella typhimurium

Science.gov (United States)

Wu, Wen-he; Li, Min; Wang, Yue; Ouyang, Hou-xian; Wang, Lin; Li, Ci-xiu; Cao, Yu-chen; Meng, Qing-he; Lu, Jian-xin

2012-11-01

Herein we reported the development of aptamer-based biosensors (aptasensors) based on label-free aptamers and gold nanoparticles (AuNPs) for detection of Escherichia coli ( E. coli) O157:H7 and Salmonella typhimurium. Target bacteria binding aptamers are adsorbed on the surface of unmodified AuNPs to capture target bacteria, and the detection was accomplished by target bacteria-induced aggregation of the aptasensor which is associated as red-to-purple color change upon high-salt conditions. By employing anti- E. coli O157:H7 aptamer and anti- S. typhimurium aptamer, we developed a convenient and rapid approach that could selectively detect bacteria without specialized instrumentation and pretreatment steps such as cell lysis. The aptasensor could detect as low as 105colony-forming units (CFU)/ml target bacteria within 20 min or less and its specificity was 100%. This novel method has a great potential application in rapid detection of bacteria in the near future.

Interactions of Salmonella enterica Serovar Typhimurium and Pectobacterium carotovorum within a Tomato Soft Rot.

Science.gov (United States)

George, Andrée S; Cox, Clayton E; Desai, Prerak; Porwollik, Steffen; Chu, Weiping; de Moraes, Marcos H; McClelland, Michael; Brandl, Maria T; Teplitski, Max

2018-03-01

Salmonella spp. are remarkably adaptable pathogens, and this adaptability allows these bacteria to thrive in a variety of environments and hosts. The mechanisms with which these pathogens establish within a niche amid the native microbiota remain poorly understood. Here, we aimed to uncover the mechanisms that enable Salmonella enterica serovar Typhimurium strain ATCC 14028 to benefit from the degradation of plant tissue by a soft rot plant pathogen, Pectobacterium carotovorum The hypothesis that in the soft rot, the liberation of starch (not utilized by P. carotovorum ) makes this polymer available to Salmonella spp., thus allowing it to colonize soft rots, was tested first and proven null. To identify the functions involved in Salmonella soft rot colonization, we carried out transposon insertion sequencing coupled with the phenotypic characterization of the mutants. The data indicate that Salmonella spp. experience a metabolic shift in response to the changes in the environment brought on by Pectobacterium spp. and likely coordinated by the csrBC small regulatory RNA. While csrBC and flhD appear to be of importance in the soft rot, the global two-component system encoded by barA sirA (which controls csrBC and flhDC under laboratory conditions) does not appear to be necessary for the observed phenotype. Motility and the synthesis of nucleotides and amino acids play critical roles in the growth of Salmonella spp. in the soft rot. IMPORTANCE Outbreaks of produce-associated illness continue to be a food safety concern. Earlier studies demonstrated that the presence of phytopathogens on produce was a significant risk factor associated with increased Salmonella carriage on fruits and vegetables. Here, we genetically characterize some of the requirements for interactions between Salmonella and phytobacteria that allow Salmonella spp. to establish a niche within an alternate host (tomato). Pathways necessary for nucleotide synthesis, amino acid synthesis, and motility

Safety and immunogenicity of Salmonella typhimurium expressing C-terminal truncated human IL-2 in a murine model

Directory of Open Access Journals (Sweden)

Brent Sorenson

2010-03-01

Full Text Available Brent Sorenson, Kaysie Banton, Lance Augustin, Sean Barnett, Karen McCulloch, Joshua Dorn, Natalie Frykman, Arnold Leonard, Daniel SaltzmanDepartment of Surgery, University of Minnesota Medical School, Minneapolis, MN, USAAbstract: Salmonella enterica serovar Typhimurium preferentially colonizes tumors in vivo and has proven to be an effective biologic vector. The attenuated S. enterica Typhimurium strain χ4550 was engineered to express truncated human interleukin-2 and renamed SalpIL2. Previously, we observed that a single oral administration of SalpIL2 reduced tumor number and volume, while significantly increasing local and systemic natural killer (NK cell populations in an experimental metastasis model. Here we report that in nontumor-bearing mice, a single oral dose of SalpIL2 resulted in increased splenic cytotoxic T and NK cell populations that returned to control levels by 4 weeks post oral administration. Though SalpIL2 was detected in mouse tissues for up to 10 weeks, no prolonged alterations in peripheral blood serum chemistry or complete blood cell counts were observed. Similarly, comparative histopathological analysis of tissues revealed no significant increase in pyogranulomas in SalpIL2-treated animals with respect to saline controls. In Rag-1 knockout mice, which have severely impaired B and T cell function, SalpIL2 reduced growth of hepatic metastases. Furthermore, SalpIL2 altered expression of several proinflammatory cytokines and chemokines in the serum of mice with pulmonary osteosarcoma metastases. These data further suggest that SalpIL2 is avirulent and induces a cell-mediated antitumor response.Keywords: Salmonella Typhimurium, natural killer cells, interleukin-2

Laboratory surveillance of Salmonella enterica from human clinical cases in Colombia 2005-2011.

Science.gov (United States)

Rodríguez, Edna Catering; Díaz-Guevara, Paula; Moreno, Jaime; Bautista, Adriana; Montaño, Lucy; Realpe, María Elena; Della Gaspera, Anabella; Wiesner, Magdalena

Salmonella is an enteropathogen acquired through contaminated food or water. In Colombia, Salmonella spp. is included in the national surveillance of Acute Diarrhoeal Diseases and typhoid fever initiated in 1997. This report shows the phenotype and genotype results obtained from 2005 to 2011. A total of 4010 isolates of Salmonella enterica were analysed by serotyping with Kauffmann-White-LeMinor, antimicrobial resistance patterns, and pulse-field gel electrophoresis (PFGE). A total of 93 serovars were identified, of which, Typhimurium, Enteritidis, Typhi, Dublin, Panama, Derby, Braenderup, Saintpaul, and Uganda were prominent. The highest levels of resistance were found for tetracycline and nalidixic acid. Susceptibility was observed in 52.4% (2101/4010) of the isolates. Multi-resistance was recorded in 54.9% of Typhimurium isolates, with 81 different combinations. Using PFGE, 51.9% (2083/4010) isolates were analysed in 34 serovars, and 828 electrophoretic patterns were obtained. From these, 8 patterns were found in at least two Latin-American countries. The surveillance of Salmonella spp. provides information on the serovar distribution, antimicrobial resistance, and clonal distribution in Colombia, as well as information to treat this disease and control the spread of antimicrobial bacterial resistance. Copyright © 2016 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Actin restructuring during Salmonella typhimurium infection investigated by confocal and super-resolution microscopy

Science.gov (United States)

Han, Jason J.; Kunde, Yuliya A.; Hong-Geller, Elizabeth; Werner, James H.

2014-01-01

We have used super-resolution optical microscopy and confocal microscopy to visualize the cytoskeletal restructuring of HeLa cells that accompanies and enables Salmonella typhimurium internalization. Herein, we report the use of confocal microscopy to verify and explore infection conditions that would be compatible with super-resolution optical microscopy, using Alexa-488 labeled phalloidin to stain the actin cytoskeletal network. While it is well known that actin restructuring and cytoskeletal rearrangements often accompany and assist in bacterial infection, most studies have employed conventional diffraction-limited fluorescence microscopy to explore these changes. Here we show that the superior spatial resolution provided by single-molecule localization methods (such as direct stochastic optical reconstruction microscopy) enables more precise visualization of the nanoscale changes in the actin cytoskeleton that accompany bacterial infection. In particular, we found that a thin (100-nm) ring of actin often surrounds an invading bacteria 10 to 20 min postinfection, with this ring being transitory in nature. We estimate that a few hundred monofilaments of actin surround the S. typhimurium in this heretofore unreported bacterial internalization intermediate.

Incidence of Salmonella contamination in broiler chickens in Saskatchewan.

Science.gov (United States)

Bhargava, K K; O'Neil, J B; Prior, M G; Dunkelgod, K E

1983-01-01

The incidence of Salmonella contamination in ten Saskatchewan broiler flocks varying in size from 6 200 to 14 000 was investigated from February, 1977 to April, 1979. Prior to the initial chick placement, brooding equipment, feed, water and fresh litter samples were found to be free of Salmonellae. Samples obtained from the clean and disinfected processing plant equipment before the commencement of daily operation were negative except the isolation for Salmonella anatum from the fingers of the defeathering machine in flock 4. There was no evidence of Salmonella contamination in flocks 5, 6, 8 and 10. The incidence of Salmonella was lower when cloacal swabs were taken from day old chicks fasted for 48 hours than for the same groups of chicks when carcasses were blended in nutrient broth (flocks 7 and 9). The blending of such chicks appears to be a more critical test. The serotypes isolated from eviscerated birds were the same as those isolated from used litter samples. Salmonella saintpaul was isolated from a water sample at 53 days in flock 1 and the same serotype was recovered from the intestinal contents and skin of eviscerated birds. Salmonella typhimurium was recovered from the eviscerated birds and neck samples in flock 3. In flock 4, S. saintpaul and S. anatum were isolated from 13% of the eviscerated birds sampled. Salmonella thompson, Salmonella agona and Salmonella heidelberg were recovered from 61%, 5% and 1%, respectively, of the processed carcasses sampled in flock 7.

Isolamento de Salmonella enterica em gambás (Didelphis aurita e Didelphis albiventris do Estado de São Paulo, Brasil Isolation of Salmonella enterica in opossum (Didelphis aurita and Didelphis albiventris of the São Paulo State, Brazil

Directory of Open Access Journals (Sweden)

Renata Assis Casagrande

2011-03-01

Full Text Available No Brasil, não há relato de estudos de Salmonella em gambás, sendo assim, este trabalho tem por objetivo determinar a frequência de isolamento de Salmonella enterica em gambás (D. aurita e D. albiventris no Estado de São Paulo. No período de janeiro de 2005 a dezembro de 2006, foram necropsiados 106 D. aurita e 40 D. albiventris e colhidos fragmentos de intestinos delgado, grosso e suabe da cloaca. As amostras foram plaqueadas diretamente em ágar Mac Conkey, paralelamente suspendidas nos caldos Rappaport-Vassiliadis e Tetrationato e posteriormente plaqueados em ágar XLT4. As colônias sugestivas de Salmonella foram confirmadas através de provas bioquímicas e sorotipagem. Encontrou-se Salmonella enterica em 17,0% (18/106 dos D. aurita. Destes, 50% apresentaram positividade no intestino delgado (ID, 88,9% no intestino grosso (IG e 66,7% na cloaca. Da espécie S. enterica, as subespécies encontradas foram: diarizonae (11,1% houtenae e enterica (5,5% cada um; enquanto da subespécie S. enterica enterica os sorotipos foram Newport (83,3%, Typhimurium e Cerro (5,5% cada um. Nos D. albiventris, 17,5% (7/40 eram positivos, sendo que se encontraram 42,8% no ID, 85,7% no IG e 71,4% na cloaca. O sorotipo mais prevalente também foi Newport (71,4%, seguido por Typhimurium, Bareilly e Thompson (14,3% cada um. Através dos resultados obtidos neste estudo pode-se comprovar a presença de Salmonella enterica no trato intestinal de gambás no Brasil.In Brazil there is not report of Salmonella in opossum, so then, the objective of this study is to determine the isolation frequency of Salmonella enterica in opossum in São Paulo State, Brazil. From January 2005 to December 2006, 106 D. aurita and 40 D. albiventris were necropsied and samples from small and large intestine and cloacal swab were collected. These samples were submitted to direct plating in Mac Conkey agar and parallel suspension in Rappaport-Vassiliadis and Tetrationate broths with

Non-Saccharomyces yeasts protect against epithelial cell barrier disruption induced by Salmonella enterica subsp. enterica serovar Typhimurium.

Science.gov (United States)

Smith, I M; Baker, A; Arneborg, N; Jespersen, L

2015-11-01

The human gastrointestinal epithelium makes up the largest barrier separating the body from the external environment. Whereas invasive pathogens cause epithelial barrier disruption, probiotic micro-organisms modulate tight junction regulation and improve epithelial barrier function. In addition, probiotic strains may be able to reduce epithelial barrier disruption caused by pathogenic species. The aim of this study was to explore non-Saccharomyces yeast modulation of epithelial cell barrier function in vitro. Benchmarking against established probiotic strains, we evaluated the ability of four nonpathogenic yeast species to modulate transepithelial electrical resistance (TER) across a monolayer of differentiated human colonocytes (Caco-2 cells). Further, we assessed yeast modulation of a Salmonella Typhimurium-induced epithelial cell barrier function insult. Our findings demonstrate distinct patterns of non-Saccharomyces yeast modulation of epithelial cell barrier function. While the established probiotic yeast Saccharomyces boulardii increased TER across a Caco-2 monolayer by 30%, Kluyveromyces marxianus exhibited significantly stronger properties of TER enhancement (50% TER increase). In addition, our data demonstrate significant yeast-mediated modulation of Salmonella-induced epithelial cell barrier disruption and identify K. marxianus and Metschnikowia gruessii as two non-Saccharomyces yeasts capable of protecting human epithelial cells from pathogen invasion. This study demonstrates distinct patterns of non-Saccharomyces yeast modulation of epithelial cell barrier function in vitro. Further, our data demonstrate significant yeast-mediated modulation of Salmonella Typhimurium-induced epithelial cell barrier disruption and identify Kluyveromyces marxianus and Metschnikowia gruessii as two non-Saccharomyces yeasts capable of protecting human epithelial cells from pathogen invasion. This study is the first to demonstrate significant non-Saccharomyces yeast

Sensibilidade e resistência de amostras de Salmonella Typhimurium isoladas de suínos abatidos no Rio Grande do Sul/Brasil frente aos desinfetantes químicos quaternário de amônio e iodofor Sensitivity and resistance of samples of Salmonella Typhimurium isolated in slaughter swines in the state Rio Grande do Sul/Brazil, front to disinfectants quaternary ammonium and iodophor

Directory of Open Access Journals (Sweden)

Luciane Martins Borowsky

2006-10-01

of 96 Salmonella Typhimurium strains isolated from slaughter pigs in the state of Rio Grande do Sul, Brazil. The isolates were tested against quaternary ammonium and iodophor, which represent two commercial disinfectants commonly used in animal production. The tested disinfectants were used in the concentration recommended by the fabricant and in a sub-concentration in order to simulate a possible field situation. Dilution suspension tests were conducted, observing the inactivation of each S.Typhimurium isolate after 5, 15, 30 and 60 minutes of contact with each compound. All tested isolates were inactivated by the quaternary ammonium compound in both concentrations. Four isolates revealed resistant to iodophor in the recommended concentration and 59 isolates when a sub-concentration was tested. The testing of resistance against disinfectants in microorganisms present on farm and in food processing plants might be an important step on monitoring the effectiveness of adopted disinfections protocols.

Efecto de la exposición a la luz ultravioleta uv-c en la viabilidad de especies de Eschericha coli y Salmonella typhimurium

OpenAIRE

Oviedo, Dumas; Rojas, Jesús María; Borda, Ricardo Alberto; Durango, Mónica María

2013-01-01

Introduction. The germicidal effect UV-C light has is regarded as an effective tool to inactivate and eliminate harmful contaminating agents, such as Escherichia coli and Salmonella typhimurium. Objective. evaluating the effectiveness of UV-C light for reducing Escherichia coli and Salmonella typhimorium populations from cultures that had the microorganisms, combining factors like concentration, time and distance. Methodology. A UV-C lamp, with a 254 nm and 8 ...

Antibacterial activity of Ziziphora clinopodioides essential oil and nisin against Bacillus subtilis and Salmonella Typhimurium in commercial barley soup

Directory of Open Access Journals (Sweden)

Y. Shahbazi

2017-03-01

Full Text Available The objective of the current study was to evaluate the antibacterial activity of nisin (250 and 500 IU/mL, Ziziphora clinopodioides essential oil (0.1 and 0.2% and their combination against Bacillus subtilis and Salmonella Typhimurium in commercial barley soup during refrigerated storage. Based on our findings, the population of pathogens was reduced with the addition of the essential oil and nisin, increased concentration of the added antibacterial agents and the longer storage time of commercial barley soup. The group treated with the essential oil at 0.2% and nisin at 500 IU/mL showed the most rapid decrease in the number of S. Typhimurium and B. subtilis. By the end of seven and five days, populations of S. Typhimurium and B. subtilis in overall concentrations of the essential oil with nisin were totally inhibited, respectively. Our findings suggest the possibility of utilising commercial barley soup with a mixture of Z. clinopodioides essential oil and nisin for the reduction of B. subtilis and S. Typhimurium.

Molecular epidemiology of fluoroquinolone resistant Salmonella in Africa: A systematic review and meta-analysis.

Science.gov (United States)

Tadesse, Getachew; Tessema, Tesfaye S; Beyene, Getenet; Aseffa, Abraham

2018-01-01

Wide-ranging evidence on the occurrence of fluoroquinolone (FQ) resistance genetic determinants in African Salmonella strains is not available. The main objectives of this study were to assess the heterogeneity, estimate pooled proportions and describe the preponderance of FQ-resistance determinants in typhoidal and non-typhoidal Salmonella (NTS) isolates of Africa. Genetic and phenotypic data on 6103 Salmonella isolates were considered. Meta- and frequency analyses were performed depending on the number of studies by category, number of isolates and risks of bias. A random effects model was used to assess heterogeneity and estimate pooled proportions. Relative and cumulative frequencies were calculated to describe the overall preponderance of FQ-resistance determinants in quinolone resistant isolates. The pooled proportion of gyrA mutants (Salmonella enterica serovar Typhi, Salmonella enterica serovar Typhimurium, and Salmonella enterica serovar Enteritidis) was estimated at 5.7% (95% Confidence interval (CI) = 2.6, 9.8; Tau squared (T2) = 0.1105), and was higher in S. Typhi than in S. Typhimurium (odds ratio (OR) = 3.3, 95%CI = 2, 5.7). The proportions of each of gyrB and parC mutants, and strains with Plasmid Mediated Quinolone Resistance genes (qnrA, qnrB and qnrS) were low (≤ 0.3%). Overall, 23 mutant serotypes were identified, and most strains had mutations at codons encoding Ser83 and Asp87 of gyrA (82%, 95%CI = 78, 86). Mutations at gyrA appear to account for ciprofloxacin non-susceptibility in most clinical Salmonella strains in Africa. The estimates could be harnessed to develop a mismatch-amplification mutation-assay for the detection of FQ-resistant strains in Africa.

Changes in thermo-tolerance and survival under simulated gastrointestinal conditions of Salmonella Enteritidis PT4 and Salmonella Typhimurium PT4 in chicken breast meat after exposure to sequential stresses.

Science.gov (United States)

Melo, Adma Nadja Ferreira de; Souza, Geany Targino de; Schaffner, Donald; Oliveira, Tereza C Moreira de; Maciel, Janeeyre Ferreira; Souza, Evandro Leite de; Magnani, Marciane

2017-06-19

This study assessed changes in thermo-tolerance and capability to survive to simulated gastrointestinal conditions of Salmonella Enteritidis PT4 and Salmonella Typhimurium PT4 inoculated in chicken breast meat following exposure to stresses (cold, acid and osmotic) commonly imposed during food processing. The effects of the stress imposed by exposure to oregano (Origanum vulgare L.) essential oil (OVEO) on thermo-tolerance were also assessed. After exposure to cold stress (5°C for 5h) in chicken breast meat the test strains were sequentially exposed to the different stressing substances (lactic acid, NaCl or OVEO) at sub-lethal amounts, which were defined considering previously determined minimum inhibitory concentrations, and finally to thermal treatment (55°C for 30min). Resistant cells from distinct sequential treatments were exposed to simulated gastrointestinal conditions. The exposure to cold stress did not result in increased tolerance to acid stress (lactic acid: 5 and 2.5μL/g) for both strains. Cells of S. Typhimurium PT4 and S. Enteritidis PT4 previously exposed to acid stress showed higher (pthermo-tolerance in both strains. The cells that survived the sequential stress exposure (resistant) showed higher tolerance (pthermo-tolerance and enhance the survival under gastrointestinal conditions of S. Enteritidis PT4 and S. Typhimurium PT4. Copyright © 2017 Elsevier B.V. All rights reserved.

Attachment behaviour of Escherichia coli K12 and Salmonella Typhimurium P6on food contact surfaces for food transportation

DEFF Research Database (Denmark)

Abban, Stephen; Jakobsen, Mogens; Jespersen, Lene

2012-01-01

The role of cargo container lining materials aluminium, a fibre reinforced plastic (FRP) and stainless steel in bacterial cross contamination during transport was assessed. For this, attachment and detachment of Escherichia coli K12 and Salmonella Typhimurium P6 on the three surfaces in the absence....... Typhimurium P6 respectively. Correlation with roughness average was poor; r = -0.425 and -0.413 respectively for E. coli K12 and S. Typhimurium P6. Presence of residue caused significant reduction (p ... material sections of the same surfaces. We report these observations for the first time for aluminium and the FRP material and in part for stainless steel. The S. Typhimurium P6 strain also had significantly higher level of attachment than the E. coli K12 strain. Our findings show that food residue...

Pediatric Epidemic of Salmonella enterica Serovar Typhimurium in the Area of L’Aquila, Italy, Four Years after a Catastrophic Earthquake

Directory of Open Access Journals (Sweden)

Giovanni Nigro

2016-05-01

Full Text Available Background: A Salmonella enterica epidemic occurred in children of the area of L’Aquila (Central Italy, Abruzzo region between June 2013 and October 2014, four years after the catastrophic earthquake of 6 April 2009. Methods: Clinical and laboratory data were collected from hospitalized and ambulatory children. Routine investigations for Salmonella infection were carried out on numerous alimentary matrices of animal origin and sampling sources for drinking water of the L’Aquila district, including pickup points of the two main aqueducts. Results: Salmonella infection occurred in 155 children (83 females: 53%, aged 1 to 15 years (mean 2.10. Of these, 44 children (28.4% were hospitalized because of severe dehydration, electrolyte abnormalities, and fever resistant to oral antipyretic and antibiotic drugs. Three children (1.9% were reinfected within four months after primary infection by the same Salmonella strain. Four children (2.6%, aged one to two years, were coinfected by rotavirus. A seven-year old child had a concomitant right hip joint arthritis. The isolated strains, as confirmed in about the half of cases or probable/possible in the remaining ones, were identified as S. enterica serovar Typhimurium [4,5:i:-], monophasic variant. Aterno river, bordering the L’Aquila district, was recognized as the main responsible source for the contamination of local crops and vegetables derived from polluted crops. Conclusions: The high rate of hospitalized children underlines the emergence of a highly pathogenic S. enterica strain probably subsequent to the contamination of the spring water sources after geological changes occurred during the catastrophic earthquake.

Pediatric Epidemic of Salmonella enterica Serovar Typhimurium in the Area of L'Aquila, Italy, Four Years after a Catastrophic Earthquake.

Science.gov (United States)

Nigro, Giovanni; Bottone, Gabriella; Maiorani, Daniela; Trombatore, Fabiana; Falasca, Silvana; Bruno, Gianfranco

2016-05-06

A Salmonella enterica epidemic occurred in children of the area of L'Aquila (Central Italy, Abruzzo region) between June 2013 and October 2014, four years after the catastrophic earthquake of 6 April 2009. Clinical and laboratory data were collected from hospitalized and ambulatory children. Routine investigations for Salmonella infection were carried out on numerous alimentary matrices of animal origin and sampling sources for drinking water of the L'Aquila district, including pickup points of the two main aqueducts. Salmonella infection occurred in 155 children (83 females: 53%), aged 1 to 15 years (mean 2.10). Of these, 44 children (28.4%) were hospitalized because of severe dehydration, electrolyte abnormalities, and fever resistant to oral antipyretic and antibiotic drugs. Three children (1.9%) were reinfected within four months after primary infection by the same Salmonella strain. Four children (2.6%), aged one to two years, were coinfected by rotavirus. A seven-year old child had a concomitant right hip joint arthritis. The isolated strains, as confirmed in about the half of cases or probable/possible in the remaining ones, were identified as S. enterica serovar Typhimurium [4,5:i:-], monophasic variant. Aterno river, bordering the L'Aquila district, was recognized as the main responsible source for the contamination of local crops and vegetables derived from polluted crops. The high rate of hospitalized children underlines the emergence of a highly pathogenic S. enterica strain probably subsequent to the contamination of the spring water sources after geological changes occurred during the catastrophic earthquake.

Effects of dietary alfalfa inclusion on Salmonella typhimurium populations in growing layer chicks.

Science.gov (United States)

Escarcha, Jacquelyn F; Callaway, Todd R; Byrd, J Allen; Miller, Dan N; Edrington, Tom S; Anderson, Robin C; Nisbet, David J

2012-10-01

Reducing Salmonella in poultry has been a paramount goal of the poultry industry in order to improve food safety. Inclusion of high-fiber fermentable feedstuffs in chicken diets has been shown to reduce the incidence of Salmonella colonization in laying hens, but no work has been performed in growing birds. Therefore, the present study was designed to quantify differences in artificially inoculated cecal Salmonella Typhimurium populations in growing layer chicks (n = 60 in each of two replications) fed 0%, 25%, and 50% of their diet (w/w) replaced with alfalfa meal from day (d) 7 to d14 after hatch. Alfalfa supplementation reduced cecal populations of Salmonella by 0.95 and 1.25 log10 colony-forming unit per gram in the 25% and 50% alfalfa groups compared to controls. Alfalfa feeding reduced (p < 0.05) the number of cecal- and crop-positive birds compared to controls. Increasing levels of alfalfa increased (p < 0.05) total volatile fatty acids (VFA) and the proportion of acetate in the cecum. Surprisingly, alfalfa inclusion did not negatively impact average daily gain (ADG) in birds over the 7-d feeding period. Alfalfa inclusion at 50% of the diet increased (p < 0.05) the number of bacterial genera detected in the cecum compared to controls, and also altered proportions of the microbial population by reducing Ruminococcus and increasing Clostridia populations. Results support the idea that providing a fermentable substrate can increase gastrointestinal VFA production and bacterial diversity which in turn can reduce colonization by Salmonella via natural competitive barriers. However, further studies are obviously needed to more fully understand the impact of changes made in diet or management procedures on poultry production.

Prevalence and epidemiology of Salmonella enterica serovar Gallinarum from poultry in some parts of Haryana, India

Directory of Open Access Journals (Sweden)

Devan Arora

2015-11-01

Full Text Available Aim: The present study was investigated to ascertain the epidemiological status of fowl typhoid (FT in broilers in some parts of Haryana during January 2011 to December 2013. Materials and Methods: To elucidate the epidemiological status of FT in broiler chickens for the 3 years (2011-2013 and to study the prevalence of various Salmonella serovars in poultry on the basis of culture characteristics, biochemical features, serotyping, and their antibiogram profile from some parts of Haryana (India. Results: A total of 309 outbreaks of FT were recorded in chickens during this period. Overall percent morbidity, mortality, case-fatality rate (CFR in broiler chicks due to FT during this period was 9.45, 6.77, and 71.55. The yearly observations were divided into quarters A (January-March, B (April-June, C (July-September and D (October-December. Maximum number of outbreaks - 106 (34.3% was recorded in quarter D followed by quarters B - 84 (27.3%, C - 64 (20.7%, and A - 55 (17.7%. Salmonella isolates (253 were recovered from disease outbreaks in broilers from different parts of Haryana. Typical morphology and colony characters on MacConkeys Lactose Agar and Brilliant Green agar, biochemical reactions, serotyping along with antibiogram profiles were able to group these isolates into 3 groups namely Salmonella Gallinarum (183, Salmonella Enteritidis (41 and Salmonella Typhimurium (29. The antibiogram pattern of 183 isolates of S. Gallinarum revealed that most of the isolates were sensitive to gentamicin (76% followed by amikacin (72%, kanamycin (71%. Conclusion: FT is prevalent in commercial broiler flocks in different parts of Haryana and is responsible for considerably high morbidity and mortality in affected flocks. Isolation of S. Gallinarum (9, 12:183 from FT cases suggest it to be the primary pathogen, however, isolation of S. Typhimurium and S. Enteritidis from these cases is a major concern. The detection of S. Enteritidis and S. Typhimurium from

Molecular Epidemiology of Nontyphoidal Salmonella in Poultry and Poultry Products in India: Implications for Human Health.

Science.gov (United States)

Saravanan, Sellappan; Purushothaman, Venketaraman; Murthy, Thippichettypalayam Ramasamy Gopala Krishna; Sukumar, Kuppannan; Srinivasan, Palani; Gowthaman, Vasudevan; Balusamy, Mohan; Atterbury, Robert; Kuchipudi, Suresh V

2015-09-01

Human infections with non-typhoidal Salmonella (NTS) serovars are increasingly becoming a threat to human health globally. While all motile Salmonellae have zoonotic potential, Salmonella Enteritidis and Salmonella Typhimurium are most commonly associated with human disease, for which poultry are a major source. Despite the increasing number of human NTS infections, the epidemiology of NTS in poultry in India has not been fully understood. Hence, as a first step, we carried out epidemiological analysis to establish the incidence of NTS in poultry to evaluate the risk to human health. A total of 1215 samples (including poultry meat, tissues, egg and environmental samples) were collected from 154 commercial layer farms from southern India and screened for NTS. Following identification by cultural and biochemical methods, Salmonella isolates were further characterized by multiplex PCR, allele-specific PCR, enterobacterial repetitive intergenic consensus (ERIC) PCR and pulse field gel electrophoresis (PFGE). In the present study, 21/1215 (1.73 %) samples tested positive for NTS. We found 12/392 (3.06 %) of tissue samples, 7/460 (1.52 %) of poultry products, and 2/363 (0.55 %) of environmental samples tested positive for NTS. All the Salmonella isolates were resistant to oxytetracycline, which is routinely used as poultry feed additive. The multiplex PCR results allowed 16/21 isolates to be classified as S. Typhimurium, and five isolates as S. Enteritidis. Of the five S. Enteritidis isolates, four were identified as group D Salmonella by allele-specific PCR. All of the isolates produced different banding patterns in ERIC PCR. Of the thirteen macro restriction profiles (MRPs) obtained by PFGE, MRP 6 was predominant which included 6 (21 %) isolates. In conclusion, the findings of the study revealed higher incidence of contamination of NTS Salmonella in poultry tissue and animal protein sources used for poultry. The results of the study warrants further investigation

Effects of propolis from Brazil and Bulgaria on Salmonella serovars

Directory of Open Access Journals (Sweden)

R. O. Orsi

2007-01-01

Full Text Available Propolis shows biological properties such as antibacterial action. This bee product has a complex chemical composition, which depends on the local flora where it is produced. Salmonella serovars are responsible for human diseases that range from localized gastroenteritis to systemic infections. The aim of the present study was to investigate the susceptibility of Salmonella strains, isolated from food and infectious processes, to the antibacterial action of Brazilian and Bulgarian propolis, as well as to determine the behavior of these bacteria, according to the incubation period, in medium plus propolis. Dilution of ethanolic extract of propolis in agar was the used method. Brazilian and Bulgarian propolis showed an antibacterial action against all Salmonella serovars. The minimal inhibitory concentrations (MIC of propolis were similar, although they were collected in different geographic regions. Salmonella typhimurium, isolated from human infection, was more resistant to propolis than Salmonella enteritidis.

Coordinated Regulation of Virulence during Systemic Infection of Salmonella enterica serovar Typhimurium

Energy Technology Data Exchange (ETDEWEB)

Yoon, Hyunjin; McDermott, Jason E.; Porwollik, Steffen; Mcclelland, Michael; Heffron, Fred

2009-02-20

Salmonella must respond to a myriad of environmental cues during infection of a mouse and express specific subsets of genes in a temporal and spatial manner to subvert the host defense mechanisms but these regulatory pathways are poorly established. To unravel how micro-environmental signals are processed and integrated into coordinated action, we constructed in-frame non-polar deletions of 84 regulators inferred to play a role in Salmonella typhimurium virulence and tested them in three virulence assays (intraperitoneal (i.p.), and intragastric (i.g.) infection in BALB/c mice, and persistence in SvJ129 mice). Overall 36 regulators were identified that were less virulent in at least one assay, and of those, 15 regulators were required for systemic mouse infection in an acute infection model. As a first step towards understanding the interplay between a pathogen and its host from a systems biology standpoint we focused on these 15 genes. Transcriptional profiles were obtained for each of these 15 regulators from strains grown under four different environmental conditions. These results as well as publicly available transcriptional profiles were analyzed using both network inference and cluster analysis algorithms. The analysis predicts a regulatory network in which all 15 regulators control a specific set of genes necessary for Salmonella to cause systemic infection. We tested the regulatory model by expressing a subset of the regulators in trans and monitoring transcription of 7 known virulence factors located within Salmonella pathogenicity island 2 (SPI-2). These experiments validated the regulatory model and showed that, for these 7 genes, the response regulator SsrB and the marR type regulator SlyA co-regulate in a regulatory cascade by integrating multiple signals.

Curli Fibers Are Highly Conserved between Salmonella typhimurium and Escherichia coli with Respect to Operon Structure and Regulation

Science.gov (United States)

Römling, Ute; Bian, Zhao; Hammar, Mårten; Sierralta, Walter D.; Normark, Staffan

1998-01-01

Mouse-virulent Salmonella typhimurium strains SR-11 and ATCC 14028-1s express curli fibers, thin aggregative fibers, at ambient temperature on plates as judged by Western blot analysis and electron microscopy. Concomitantly with curli expression, cells develop a rough and dry colony morphology and bind the dye Congo red (called the rdar morphotype). Cloning and characterization of the two divergently transcribed operons required for curli biogenesis, csgBA(C) and csgDEFG, from S. typhimurium SR-11 revealed the same gene order and flanking genes as in Escherichia coli. The divergence of the curli region between S. typhimurium and E. coli at the nucleotide level is above average (22.4%). However, a high level of conservation at the protein level, which ranged from 86% amino acid homology for the fiber subunit CsgA to 99% homology for the lipoprotein CsgG, implies functional constraints on the gene products. Consequently, S. typhimurium genes on low-copy-number plasmids were able to complement respective E. coli mutants, although not always to wild-type levels. rpoS and ompR are required for transcriptional activation of (at least) the csgD promoter. The high degree of conservation at the protein level and the identical regulation patterns in E. coli and S. typhimurium suggest similar roles of curli fibers in the same ecological niche in the two species. PMID:9457880

A low-pH medium in vitro or the environment within a macrophage decreases the transcriptional levels of fimA, fimZ and lrp in Salmonella enterica serovar Typhimurium.

Science.gov (United States)

Wang, Ke-Chuan; Hsu, Yuan-Hsun; Huang, Yi-Ning; Chen, Ter-Hsin; Lin, Jiunn-Horng; Hsuan, Shih-Ling; Chien, Maw-Sheng; Lee, Wei-Cheng; Yeh, Kuang-Sheng

2013-09-01

Many Salmonella Typhimurium isolates produce type 1 fimbriae and exhibit fimbrial phase variation in vitro. Static broth culture favours the production of fimbriae, while solid agar medium inhibits the generation of these appendages. Little information is available regarding whether S. Typhimurium continues to produce type 1 fimbriae during in vivo growth. We used a type 1 fimbrial phase-variable strain S. Typhimurium LB5010 and its derivatives to infect RAW 264.7 macrophages. Following entry into macrophages, S. Typhimurium LB5010 gradually decreased the transcript levels of fimbrial subunit gene fimA, positive regulatory gene fimZ, and global regulatory gene lrp. A similar decrease in transcript levels was detected by RT-PCRwhen the pH of static brothmediumwas shifted frompH 7 to amore acidic pH 4. A fimA-deleted strain continued to multiply within macrophages as did the parental strain. An lrp deletion strain was unimpaired for in vitro growth at pH 7 or pH 4, while a strain harboring an lrp-containing plasmid exhibited impaired in vitro growth at pH 4. We propose that acidic medium, which resembles one aspect of the intracellular environment in a macrophage, inhibits type 1 fimbrial production by down-regulation of the expression of lrp, fimZ and fimA.

Saccharomyces boulardii modifies Salmonella typhimurium traffic and host immune responses along the intestinal tract.

Directory of Open Access Journals (Sweden)

Rodolphe Pontier-Bres

Full Text Available Salmonella enterica serovar Typhimurium (ST is an enteropathogenic Gram-negative bacterium that causes infection following oral ingestion. ST spreads rapidly along the gastrointestinal tract (GIT and invades the intestinal epithelium to ultimately reach internal body organs. The probiotic yeast Saccharomyces boulardii BIOCODEX (S.b-B is prescribed for prophylaxis of diarrheal infectious diseases. We previously showed that S.b-B prevents weight loss in ST-infected mice and significantly decreases bacterial translocation to the spleen and liver. This study was designed to investigate the effect of S.b-B on ST migration along the GIT and the impact of the yeast on the host's early innate immune responses. Bioluminescent imaging (BLI was used to evaluate the effect of S.b-B on the progression of luminescent Salmonella Typhimurium (ST-lux in the GIT of mice pretreated with streptomycin. Photonic emission (PE was measured in GIT extracts (stomach, small intestine, cecum and colon at various time periods post-infection (PI. PE analysis revealed that, 45 min PI, ST-lux had migrated slightly faster in the mice treated with S.b-B than in the untreated infected animals. At 90 min PI, ST-lux had reached the cecum in both groups of mice. Adhesion of ST to S.b-B was visualized in the intestines of the mice and probably accounts for (1 the faster elimination of ST-lux in the feces, and (2 reduced translocation of ST to the spleen and liver. In the early phase of infection, S.b-B also modifies the host's immune responses by (1 increasing IFN-γ gene expression and decreasing IL-10 gene expression in the small intestine, and (2 elevating both IFN-γ, and IL-10 mRNA levels in the cecum. BLI revealed that S.b-B modifies ST migration and the host immune response along the GIT. Study findings shed new light on the protective mechanisms of S.b-B during the early phase of Salmonella pathogenesis.

Saccharomyces boulardii modifies Salmonella typhimurium traffic and host immune responses along the intestinal tract.

Science.gov (United States)

Pontier-Bres, Rodolphe; Munro, Patrick; Boyer, Laurent; Anty, Rodolphe; Imbert, Véronique; Terciolo, Chloé; André, Fréderic; Rampal, Patrick; Lemichez, Emmanuel; Peyron, Jean-François; Czerucka, Dorota

2014-01-01

Salmonella enterica serovar Typhimurium (ST) is an enteropathogenic Gram-negative bacterium that causes infection following oral ingestion. ST spreads rapidly along the gastrointestinal tract (GIT) and invades the intestinal epithelium to ultimately reach internal body organs. The probiotic yeast Saccharomyces boulardii BIOCODEX (S.b-B) is prescribed for prophylaxis of diarrheal infectious diseases. We previously showed that S.b-B prevents weight loss in ST-infected mice and significantly decreases bacterial translocation to the spleen and liver. This study was designed to investigate the effect of S.b-B on ST migration along the GIT and the impact of the yeast on the host's early innate immune responses. Bioluminescent imaging (BLI) was used to evaluate the effect of S.b-B on the progression of luminescent Salmonella Typhimurium (ST-lux) in the GIT of mice pretreated with streptomycin. Photonic emission (PE) was measured in GIT extracts (stomach, small intestine, cecum and colon) at various time periods post-infection (PI). PE analysis revealed that, 45 min PI, ST-lux had migrated slightly faster in the mice treated with S.b-B than in the untreated infected animals. At 90 min PI, ST-lux had reached the cecum in both groups of mice. Adhesion of ST to S.b-B was visualized in the intestines of the mice and probably accounts for (1) the faster elimination of ST-lux in the feces, and (2) reduced translocation of ST to the spleen and liver. In the early phase of infection, S.b-B also modifies the host's immune responses by (1) increasing IFN-γ gene expression and decreasing IL-10 gene expression in the small intestine, and (2) elevating both IFN-γ, and IL-10 mRNA levels in the cecum. BLI revealed that S.b-B modifies ST migration and the host immune response along the GIT. Study findings shed new light on the protective mechanisms of S.b-B during the early phase of Salmonella pathogenesis.

Genomic characterisation of invasive non-typhoidal Salmonella enterica Subspecies enterica Serovar Bovismorbificans isolates from Malawi.

Directory of Open Access Journals (Sweden)

Christina Bronowski

2013-11-01

Full Text Available Invasive Non-typhoidal Salmonella (iNTS are an important cause of bacteraemia in children and HIV-infected adults in sub-Saharan Africa. Previous research has shown that iNTS strains exhibit a pattern of gene loss that resembles that of host adapted serovars such as Salmonella Typhi and Paratyphi A. Salmonella enterica serovar Bovismorbificans was a common serovar in Malawi between 1997 and 2004.We sequenced the genomes of 14 Malawian bacteraemia and four veterinary isolates from the UK, to identify genomic variations and signs of host adaptation in the Malawian strains.Whole genome phylogeny of invasive and veterinary S. Bovismorbificans isolates showed that the isolates are highly related, belonging to the most common international S. Bovismorbificans Sequence Type, ST142, in contrast to the findings for S. Typhimurium, where a distinct Sequence Type, ST313, is associated with invasive disease in sub-Saharan Africa. Although genome degradation through pseudogene formation was observed in ST142 isolates, there were no clear overlaps with the patterns of gene loss seen in iNTS ST313 isolates previously described from Malawi, and no clear distinction between S. Bovismorbificans isolates from Malawi and the UK. The only defining differences between S. Bovismorbificans bacteraemia and veterinary isolates were prophage-related regions and the carriage of a S. Bovismorbificans virulence plasmid (pVIRBov.iNTS S. Bovismorbificans isolates, unlike iNTS S. Typhiumrium isolates, are only distinguished from those circulating elsewhere by differences in the mobile genome. It is likely that these strains have entered a susceptible population and are able to take advantage of this niche. There are tentative signs of convergent evolution to a more human adapted iNTS variant. Considering its importance in causing disease in this region, S. Bovismorbificans may be at the beginning of this process, providing a reference against which to compare changes that may

Immunogenicity of a Live Recombinant Salmonella enterica Serovar Typhimurium Vaccine Expressing pspA in Neonates and Infant Mice Born from Naïve and Immunized Mothers▿ †

OpenAIRE

Shi, Huoying; Wang, Shifeng; Roland, Kenneth L.; Gunn, Bronwyn M.; Curtiss, Roy

2010-01-01

We are developing a Salmonella vectored vaccine to prevent infant pneumonia and other diseases caused by Streptococcus pneumoniae. One prerequisite for achieving this goal is to construct and evaluate new recombinant attenuated Salmonella vaccine (RASV) strains suitable for use in neonates and infants. Salmonella enterica serovar Typhimurium strain χ9558(pYA4088) specifies delivery of the pneumococcal protective antigen PspA and can protect adult mice from challenge with S. pneumoniae. This s...

Elucidation of the outer membrane proteome of Salmonella enterica serovar Typhimurium utilising a lipid-based protein immobilization technique

Directory of Open Access Journals (Sweden)

Appleton Hazel

2010-02-01

Full Text Available Abstract Background Salmonella enterica serovar Typhimurium (S. Typhimurium is a major cause of human gastroenteritis worldwide. The outer membrane proteins expressed by S. Typhimurium mediate the process of adhesion and internalisation within the intestinal epithelium of the host thus influencing the progression of disease. Since the outer membrane proteins are surface-exposed, they provide attractive targets for the development of improved antimicrobial agents and vaccines. Various techniques have been developed for their characterisation, but issues such as carryover of cytosolic proteins still remain a problem. In this study we attempted to characterise the surface proteome of S. Typhimurium using Lipid-based Protein Immobilisation technology in the form of LPI™ FlowCells. No detergents are required and no sample clean up is needed prior to downstream analysis. The immobilised proteins can be digested with proteases in multiple steps to increase sequence coverage, and the peptides eluted can be characterised directly by liquid chromatography - tandem mass spectrometry (LC-MS/MS and identified from mass spectral database searches. Results In this study, 54 outer membrane proteins, were identified with two or more peptide hits using a multi-step digest approach. Out of these 28 were lipoproteins, nine were involved in transport and three with enzyme activity These included the transporters BtuB which is responsible for the uptake of vitamin B12, LamB which is involved in the uptake of maltose and maltodextrins and LolB which is involved in the incorporation of lipoproteins in the outer membrane. Other proteins identified included the enzymes MltC which may play a role in cell elongation and division and NlpD which is involved in catabolic processes in cell wall formation as well as proteins involved in virulence such as Lpp1, Lpp2 and OmpX. Conclusion Using a multi-step digest approach the LPI™ technique enables the incorporation of a

Effects of irradiation and fumaric acid treatment on the inactivation of Listeria monocytogenes and Salmonella typhimurium inoculated on sliced ham

Energy Technology Data Exchange (ETDEWEB)

Song, Hyeon-Jeong; Lee, Ji-Hye [Department of Food Science and Technology, Chungnam National University, Daejeon 305-764 (Korea, Republic of); Song, Kyung Bin, E-mail: kbsong@cnu.ac.kr [Department of Food Science and Technology, Chungnam National University, Daejeon 305-764 (Korea, Republic of)

2011-11-15

To examine the effects of fumaric acid and electron beam irradiation on the inactivation of foodborne pathogens in ready-to-eat meat products, sliced ham was inoculated with Listeria monocytogenes and Salmonella typhimurium. The inoculated ham slices were treated with 0.5% fumaric acid or electron beam irradiation at 2 kGy. Fumaric acid treatment reduced the populations of L. monocytogenes and S. typhimurium by approximately 1 log CFU/g compared to control populations. In contrast, electron beam irradiation decreased the populations of S. typhimurium and L. monocytogenes by 3.78 and 2.42 log CFU/g, respectively. These results suggest that electron beam irradiation is a better and appropriate technique for improving the microbial safety of sliced ham. - Highlights: > We compare irradiation and fumaric acid treatment on the inactivation of pathogens. > We examine changes in the populations of L. monocytogenes and S. typhimurium. > Irradiation at 2 kGy is more effective in sliced ham than fumaric acid treatment. > Low-dose irradiation can improve the microbial safety of sliced ham during storage.

Effects of irradiation and fumaric acid treatment on the inactivation of Listeria monocytogenes and Salmonella typhimurium inoculated on sliced ham