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Sample records for attenuated salmonella enterica

  1. Complete Genome Sequence of NC983, a Live Attenuated Strain of Salmonella enterica Serovar Typhimurium

    Science.gov (United States)

    Troxell, Bryan; Fink, Ryan C.; Dickey, Allison N.; Scholl, Elizabeth H.

    2016-01-01

    Foodborne infections caused by Salmonella enterica serovars are a significant problem worldwide. Presented here is the genome sequence of the nontyphoidal S. enterica serovar Typhimurium mutant strain NC983, a potential vaccine candidate. PMID:27738027

  2. aroA-Deficient Salmonella enterica Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant

    Science.gov (United States)

    Frahm, Michael; Kocijancic, Dino; Rohde, Manfred; Eckweiler, Denitsa; Bielecka, Agata; Bueno, Emilio; Cava, Felipe; Abraham, Wolf-Rainer; Curtiss, Roy; Häussler, Susanne; Erhardt, Marc; Weiss, Siegfried

    2016-01-01

    ABSTRACT Recombinant attenuated Salmonella enterica serovar Typhimurium strains are believed to act as powerful live vaccine carriers that are able to elicit protection against various pathogens. Auxotrophic mutations, such as a deletion of aroA, are commonly introduced into such bacteria for attenuation without incapacitating immunostimulation. In this study, we describe the surprising finding that deletion of aroA dramatically increased the virulence of attenuated Salmonella in mouse models. Mutant bacteria lacking aroA elicited increased levels of the proinflammatory cytokine tumor necrosis factor alpha (TNF-α) after systemic application. A detailed genetic and phenotypic characterization in combination with transcriptomic and metabolic profiling demonstrated that ΔaroA mutants display pleiotropic alterations in cellular physiology and lipid and amino acid metabolism, as well as increased sensitivity to penicillin, complement, and phagocytic uptake. In concert with other immunomodulating mutations, deletion of aroA affected flagellin phase variation and gene expression of the virulence-associated genes arnT and ansB. Finally, ΔaroA strains displayed significantly improved tumor therapeutic activity. These results highlight the importance of a functional shikimate pathway to control homeostatic bacterial physiology. They further highlight the great potential of ΔaroA-attenuated Salmonella for the development of vaccines and cancer therapies with important implications for host-pathogen interactions and translational medicine. PMID:27601574

  3. Salmonella enterica serovar typhimurium strains with regulated delayed attenuation in vivo.

    Science.gov (United States)

    Curtiss, Roy; Wanda, Soo-Young; Gunn, Bronwyn M; Zhang, Xin; Tinge, Steven A; Ananthnarayan, Vidya; Mo, Hua; Wang, Shifeng; Kong, Wei

    2009-03-01

    Recombinant bacterial vaccines must be fully attenuated for animal or human hosts to avoid inducing disease symptoms while exhibiting a high degree of immunogenicity. Unfortunately, many well-studied means for attenuating Salmonella render strains more susceptible to host defense stresses encountered following oral vaccination than wild-type virulent strains and/or impair their ability to effectively colonize the gut-associated and internal lymphoid tissues. This thus impairs the ability of recombinant vaccines to serve as factories to produce recombinant antigens to induce the desired protective immunity. To address these problems, we designed strains that display features of wild-type virulent strains of Salmonella at the time of immunization to enable strains first to effectively colonize lymphoid tissues and then to exhibit a regulated delayed attenuation in vivo to preclude inducing disease symptoms. We recently described one means to achieve this based on a reversible smooth-rough synthesis of lipopolysaccharide O antigen. We report here a second means to achieve regulated delayed attenuation in vivo that is based on the substitution of a tightly regulated araC P(BAD) cassette for the promoters of the fur, crp, phoPQ, and rpoS genes such that expression of these genes is dependent on arabinose provided during growth. Thus, following colonization of lymphoid tissues, the Fur, Crp, PhoPQ, and/or RpoS proteins cease to be synthesized due to the absence of arabinose such that attenuation is gradually manifest in vivo to preclude induction of diseases symptoms. Means for achieving regulated delayed attenuation can be combined with other mutations, which together may yield safe efficacious recombinant attenuated Salmonella vaccines.

  4. Immunogenicity of recombinant attenuated Salmonella enterica serovar Typhimurium vaccine strains carrying a gene that encodes Eimeria tenella antigen SO7.

    Science.gov (United States)

    Konjufca, Vjollca; Jenkins, Mark; Wang, Shifeng; Juarez-Rodriguez, Maria Dolores; Curtiss, Roy

    2008-12-01

    Recombinant attenuated Salmonella vaccines against avian coccidiosis were developed to deliver Eimeria species antigens to the lymphoid tissues of chickens via the type 3 secretion system (T3SS) and the type 2 secretion system (T2SS) of Salmonella. For antigen delivery via the T3SS, the Eimeria tenella gene encoding sporozoite antigen SO7 was cloned downstream of the translocation domain of the Salmonella enterica serovar Typhimurium sopE gene in the parental pYA3868 and pYA3870 vectors to generate pYA4156 and pYA4157. Newly constructed T3SS vectors were introduced into host strain chi8879 (Delta phoP233 Delta sptP1033::xylE Delta asdA16), an attenuated derivative of the highly virulent UK-1 strain. The SopE-SO7 fusion protein was secreted by the T3SS of Salmonella. The vector pYA4184 was constructed for delivery of the SO7 antigen via the T2SS. The SO7 protein was toxic to Salmonella when larger amounts were synthesized; thus, the synthesis of this protein was placed under the control of the lacI repressor gene, whose expression in turn was dependent on the amount of available arabinose in the medium. The pYA4184 vector was introduced into host strain chi9242 (Delta phoP233 Delta asdA16 Delta araBAD23 Delta relA198::araC P(BAD) lacI TT [TT is the T4ipIII transcription terminator]). In addition to SO7, for immunization and challenge studies we used the EAMZ250 antigen of Eimeria acervulina, which was previously shown to confer partial protection against E. acervulina challenge when it was delivered via the T3SS. Immunization of chickens with a combination of the SO7 and EAMZ250 antigens delivered via the T3SS induced superior protection against challenge by E. acervulina. In contrast, chickens immunized with SO7 that was delivered via the T2SS of Salmonella were better protected from challenge by E. tenella.

  5. Deletion of the aceE gene (encoding a component of pyruvate dehydrogenase) attenuates Salmonella enterica serovar Enteritidis.

    Science.gov (United States)

    Pang, Ervinna; Tien-Lin, Chang; Selvaraj, Madhan; Chang, Jason; Kwang, Jimmy

    2011-10-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major food-borne pathogen. From a transposon insertion mutant library created previously using S. Enteritidis 10/02, one of the mutants was identified to have a 50% lethal dose (LD(50) ) at least 100 times that of the parental strain in young chicks, with an attenuation in a poorly studied gene encoding a component of pyruvate dehydrogenase, namely the aceE gene. Evaluation of the in vitro virulence characteristics of the ΔaceE∷kan mutant revealed that it was less able to invade epithelial cells, less resistant to reactive oxygen intermediate, less able to survive within a chicken macrophage cell line and had a retarded growth rate compared with the parental strain. Young chicks vaccinated with 2 × 10(9) CFU of the ΔaceE∷kan mutant were protected from the subsequent challenge of the parental strain, with the mutant colonized in the liver and spleen in a shorter time than the group infected with the parental strain. In addition, compared with the parental strain, the ΔaceE∷kan mutant did not cause persistent eggshell contamination of vaccinated hens.

  6. Room Temperature Stabilization of Oral, Live Attenuated Salmonella enterica serovar Typhi-Vectored Vaccines

    Science.gov (United States)

    Ohtake, Satoshi; Martin, Russell; Saxena, Atul; Pham, Binh; Chiueh, Gary; Osorio, Manuel; Kopecko, Dennis; Xu, DeQi; Lechuga-Ballesteros, David; Truong-Le, Vu

    2011-01-01

    Foam drying, a modified freeze drying process, was utilized to produce a heat-stable, live attenuated Salmonella Typhi ‘Ty21a’ bacterial vaccine. Ty21a vaccine was formulated with pharmaceutically approved stabilizers, including sugars, plasticizers, amino acids, and proteins. Growth media and harvesting conditions of the bacteria were also studied to enhance resistance to desiccation stress encountered during processing as well as subsequent storage at elevated temperatures. The optimized Ty21a vaccine, formulated with trehalose, methionine, and gelatin, demonstrated stability for approximately 12 weeks at 37°C (i.e., time required for the vaccine to decrease in potency by 1log10 CFU) and no loss in titer at 4 and 25°C following storage for the same duration. Furthermore, the foam dried Ty21a elicited a similar immunogenic response in mice as well as protection in challenge studies compared to Vivotif™, the commercial Ty21a vaccine. The enhanced heat stability of the Ty21a oral vaccine, or Ty21a derivatives expressing foreign antigens (e.g. anthrax), could mitigate risks of vaccine potency loss during long term storage, shipping, delivery to geographical areas with warmer climates or during emergency distribution following a bioterrorist attack. Because the foam drying process is conducted using conventional freeze dryers and can be readily implemented at any freeze drying manufacturing facility, this technology appears ready and appropriate for large scale processing of foam dried vaccines. PMID:21300096

  7. Live attenuated vaccines for invasive Salmonella infections.

    Science.gov (United States)

    Tennant, Sharon M; Levine, Myron M

    2015-06-19

    Salmonella enterica serovar Typhi produces significant morbidity and mortality worldwide despite the fact that there are licensed Salmonella Typhi vaccines available. This is primarily due to the fact that these vaccines are not used in the countries that most need them. There is growing recognition that an effective invasive Salmonella vaccine formulation must also prevent infection due to other Salmonella serovars. We anticipate that a multivalent vaccine that targets the following serovars will be needed to control invasive Salmonella infections worldwide: Salmonella Typhi, Salmonella Paratyphi A, Salmonella Paratyphi B (currently uncommon but may become dominant again), Salmonella Typhimurium, Salmonella Enteritidis and Salmonella Choleraesuis (as well as other Group C Salmonella). Live attenuated vaccines are an attractive vaccine formulation for use in developing as well as developed countries. Here, we describe the methods of attenuation that have been used to date to create live attenuated Salmonella vaccines and provide an update on the progress that has been made on these vaccines.

  8. Oral infection with the Salmonella enterica serovar Gallinarum 9R attenuated live vaccine as a model to characterise immunity to fowl typhoid in the chicken

    Directory of Open Access Journals (Sweden)

    Beal Richard

    2005-09-01

    Full Text Available Abstract Background Salmonella enterica serovar Gallinarum (S. Gallinarum is the causative agent of fowl typhoid, a severe systemic disease of chickens that results in high mortality amongst infected flocks. Due to its virulence, the immune response to S. Gallinarum is poorly characterised. In this study we have utilised infection by the live attenuated S. Gallinarum 9R vaccine strain in inbred chickens to characterise humoral, cellular and cytokine responses to systemic salmonellosis. Results Infection with 9R results in a mild systemic infection. Bacterial clearance at three weeks post infection coincides with increases in circulating anti-Salmonella antibodies, increased T cell proliferation to Salmonella challenge and increased expression of interferon gamma. These responses peak at four weeks post infection, then decline. Only modest increases of expression of the pro-inflammatory cytokine interleukin-1β were detected early in the infection. Conclusion Infection of chickens with the 9R vaccine strain induces a mild form of systemic salmonellosis. This induces both cellular and humoral immune responses, which peak soon after bacterial clearance. Unlike enteric-associated Salmonella infections the immune response is not prolonged, reflecting the absence of persistence of Salmonella in the gastrointestinal tract. The findings here indicate that the use of the S. Gallinarum 9R vaccine strain is an effective model to study immunity to systemic salmonellosis in the chicken and may be employed in further studies to determine which components of the immune response are needed for protection.

  9. Lactic acid bacteria and bifidobacteria attenuate the proinflammatory response in intestinal epithelial cells induced by Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Carey, Christine M; Kostrzynska, Magdalena

    2013-01-01

    Inflammation is a physiological response to infections and tissue injury; however, abnormal immune responses can give rise to chronic inflammation and contribute to disease progression. Various dietary components, including probiotic lactic acid bacteria and prebiotics, have the potential to modulate intestinal inflammatory responses. One factor in particular, the chemokine interleukin-8 (IL-8, CXCL-8), is one of the major mediators of the inflammatory response. The purpose of this study was to investigate modulation of the inflammatory host response induced by Salmonella enterica serovar Typhimurium DT104 in the presence of selected probiotics and lactic acid bacteria (LAB) isolated from human sources, dairy products, and farm animals. IL-8 gene expression and protein production in HT-29 cells were evaluated by real-time PCR and ELISA, respectively. Pre-incubation of HT-29 cells with Lactobacillus kefir IM002, Bifidobacterium adolescentis FRP 61, Bifidobacterium longum FRP 68 and FRP 69, Bifidobacterium breve FRP 334, and Leuconostoc mesenteroides IM080 significantly inhibited IL-8 secretion induced by Salmonella Typhimurium DT104. Co-culture of selected probiotics and Salmonella Typhimurium DT104 reduced IL-8 production, while potential probiotics and LAB had no effect on IL-8 secretion in HT-29 cells preincubated with Salmonella Typhimurium DT104 prior to adding probiotics. Lactobacillus kefir IM002 supernatant also significantly reduced IL-8 production. In conclusion, our study suggests that probiotic bifidobacteria and LAB modulate cytokine induction and possess anti-inflammatory properties; however, the effectiveness is strain dependent.

  10. A novel Salmonella serovar isolated from Peregrine Falcon (Falco peregrinus nestlings in Sweden: Salmonella enterica enterica serovar Pajala (Salmonella Pajala

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    Jorge Hernández

    2012-08-01

    Full Text Available A novel Salmonella serovar was isolated from Peregrine falcon (Falco peregrinus nestlings in northern Sweden in 2006. Three isolates of the same clone was retrieved from three falcon siblings and characterized as Salmonella enterica sub-species enterica: O-phase 13, 23:-: e, n, z 15 and the H-phase was not present. We propose the geographical name Salmonella enterica, sub-species enterica serovar Pajala to this novel Salmonella.

  11. A novel Salmonella serovar isolated from Peregrine Falcon (Falco peregrinus) nestlings in Sweden: Salmonella enterica enterica serovar Pajala (Salmonella Pajala)

    OpenAIRE

    Hernandez, Jorge; Lindberg, Peter; Waldenström, Jonas; Drobni, Mirva; Olsen, Björn

    2012-01-01

    A novel Salmonella serovar was isolated from Peregrine falcon (Falco peregrinus) nestlings in northern Sweden in 2006. Three isolates of the same clone was retrieved from three falcon siblings and characterized as Salmonella enterica sub-species enterica: O-phase 13, 23:-: e, n, z 15 and the H-phase was not present. We propose the geographical name Salmonella enterica, sub-species entericaserovar Pajala to this novel Salmonella.Keywords: Salmonella; epidemiology; ecology; peregrine falcon; no...

  12. Prime-Boost Immunization Using a DNA Vaccine Delivered by Attenuated Salmonella enterica Serovar Typhimurium and a Killed Vaccine Completely Protects Chickens from H5N1 Highly Pathogenic Avian Influenza Virus▿

    OpenAIRE

    Pan, Zhiming; Zhang, Xiaoming; Geng, Shizhong; Fang, Qiang; You, Meng; Zhang, Lei; Jiao, Xinan; Liu, Xiufan

    2010-01-01

    H5N1 highly pathogenic avian influenza virus (HPAIV) has posed a great threat not only for the poultry industry but also for human health. However, an effective vaccine to provide a full spectrum of protection is lacking in the poultry field. In the current study, a novel prime-boost vaccination strategy against H5N1 HPAIV was developed: chickens were first orally immunized with a hemagglutinin (HA) DNA vaccine delivered by attenuated Salmonella enterica serovar Typhimurium, and boosting with...

  13. Salmonella enterica serovar typhimurium trxA mutants are protective against virulent challenge and induce less inflammation than the live-attenuated vaccine strain SL3261.

    Science.gov (United States)

    Peters, S E; Paterson, G K; Bandularatne, E S D; Northen, H C; Pleasance, S; Willers, C; Wang, J; Foote, A K; Constantino-Casas, F; Scase, T J; Blacklaws, B A; Bryant, C E; Mastroeni, P; Charles, I G; Maskell, D J

    2010-01-01

    In Salmonella enterica serovar Typhimurium, trxA encodes thioredoxin 1, a small, soluble protein with disulfide reductase activity, which catalyzes thiol disulfide redox reactions in a variety of substrate proteins. Thioredoxins are involved as antioxidants in defense against oxidative stresses, such as exposure to hydrogen peroxide and hydroxyl radicals. We have made a defined, complete deletion of trxA in the mouse-virulent S. Typhimurium strain SL1344 (SL1344 trxA), replacing the gene with a kanamycin resistance gene cassette. SL1344 trxA was attenuated for virulence in BALB/c mice by the oral and intravenous routes and when used in immunization experiments provided protection against challenge with the virulent parent strain. SL1344 trxA induced less inflammation in murine spleens and livers than SL3261, the aroA mutant, live attenuated vaccine strain. The reduced splenomegaly observed following infection with SL1344 trxA was partially attributed to a reduction in the number of both CD4(+) and CD8(+) T cells and B lymphocytes in the spleen and reduced infiltration by CD11b(+) cells into the spleen compared with spleens from mice infected with SL3261. This less severe pathological response indicates that a trxA mutation might be used to reduce reactogenicity of live attenuated vaccine strains. We tested this by deleting trxA in SL3261. SL3261 trxA was also less inflammatory than SL3261 but was slightly less effective as a vaccine strain than either the SL3261 parent strain or SL1344 trxA.

  14. Regulated delayed expression of rfaH in an attenuated Salmonella enterica serovar typhimurium vaccine enhances immunogenicity of outer membrane proteins and a heterologous antigen.

    Science.gov (United States)

    Kong, Qingke; Liu, Qing; Roland, Kenneth L; Curtiss, Roy

    2009-12-01

    RfaH is a transcriptional antiterminator that reduces the polarity of long operons encoding secreted and surface-associated cell components of Salmonella enterica serovar Typhimurium, including O antigen and lipopolysaccharide core sugars. A DeltarfaH mutant strain is attenuated in mice (50% lethal dose [LD(50)], >10(8) CFU). To examine the potential for using rfaH in conjunction with other attenuating mutations, we designed a series of strains in which we replaced the native rfaH promoter with the tightly regulated arabinose-dependent araC P(BAD) promoter so that rfaH expression was dependent on exogenously supplied arabinose provided during in vitro growth. Following colonization of host lymphoid tissues, where arabinose was not available, the P(BAD) promoter was no longer active and rfaH was not expressed. In the absence of RfaH, O antigen and core sugars were not synthesized. We constructed three mutant strains that expressed different levels of RfaH by altering the ribosome-binding sequence and start codon. One mutation, DeltaP(rfaH178), was introduced into the attenuated vaccine strain chi9241 (DeltapabA DeltapabB DeltaasdA) expressing the pneumococcal surface protein PspA from an Asd(+) balanced-lethal plasmid. Mice immunized with this strain and boosted 4 weeks later induced higher levels of serum immunoglobulin G specific for PspA and for outer membrane proteins from other enteric bacteria than either an isogenic DeltarfaH derivative or the isogenic RfaH(+) parent. Eight weeks after primary oral immunization, mice were challenged with 200 LD(50) of virulent Streptococcus pneumoniae WU2. Immunization with DeltaP(rfaH178) mutant strains led to increased levels of protection compared to that of the parent chi9241 and of a DeltarfaH derivative of chi9241.

  15. The Salmonella enterica Pan-genome

    DEFF Research Database (Denmark)

    Jacobsen, Annika; Hendriksen, Rene S.; Aarestrup, Frank Møller;

    2011-01-01

    Salmonella enterica is divided into four subspecies containing a large number of different serovars, several of which are important zoonotic pathogens and some show a high degree of host specificity or host preference. We compare 45 sequenced S. enterica genomes that are publicly available (22......, and the core and pan-genome of Salmonella were estimated to be around 2,800 and 10,000 gene families, respectively. The constructed pan-genomic dendrograms suggest that gene content is often, but not uniformly correlated to serotype. Any given Salmonella strain has a large stable core, whilst...... there is an abundance of accessory genes, including the Salmonella pathogenicity islands (SPIs), transposable elements, phages, and plasmid DNA. We visualize conservation in the genomes in relation to chromosomal location and DNA structural features and find that variation in gene content is localized in a selection...

  16. Draft Genome Sequences of Salmonella enterica subsp. enterica Serovars Typhimurium and Nottingham Isolated from Food Products

    Science.gov (United States)

    Zheng, Jie; Ayers, Sherry; Melka, David C.; Curry, Phillip E.; Payne, Justin S.; Laasri, Anna; Wang, Charles; Hammack, Thomas S.; Brown, Eric W.

    2016-01-01

    A quantitative real-time PCR (qPCR) designed to detect Salmonella enterica subsp. enterica serovar Enteritidis, targeting the sdf gene, generated positive results for S. enterica subsp. enterica serovar Typhimurium (CFSAN033950) and S. enterica subsp. enterica serovar Nottingham (CFSAN006803) isolated from food samples. Both strains show pulsed-field gel electrophoresis (PFGE) patterns distinct from those of S. Enteritidis. Here, we report the genome sequences of these two strains. PMID:27445384

  17. Virulence factors of Salmonella enterica serovar Enteritidis

    NARCIS (Netherlands)

    Zhao, Y.

    2002-01-01

    Salmonella enterica serovar Enteritidis is one of the major etiologic agents of human food-borne gastrointestinal infections. Efforts to control the number of serovar Enteritidis infections have had a limited success, in part because of the lack of knowledge of the molecular mechanisms that contribu

  18. Induction of deletion mutation onompR gene ofSalmonella enterica serovar Typhi isolates from asymptomatic typhoid carriers to evolve attenuated strains for vaccine development

    Institute of Scientific and Technical Information of China (English)

    Senthilkumar B; Anbarasu K; Senbagam D; Rajasekarapandian M

    2014-01-01

    Objective:To develop attenuated strains ofSalmonella enterica serovar Typhi(S. typhi) for the candidate vaccine by osmolar stress.Methods:S. typhiSS3 andSS5 strains were isolated from asymptomatic typhoid carriers inNamakkal,TamilNadu,India.Both strains were grown inLB (LuriaBertani) medium supplemented with various concentration ofNaCl(0.1-0.7M) respectively. The effect of osmolar stress was determined at molecular level byPCR usingMGR06 andMGR 07 primers corresponding to ompR with chromosomalDNA of S. typhiSS3 andSS5 strains. Attenuation by osmolar stress results in deletion mutation of theS. typhi strains was determined by agglutination assays, precipitation method,SDSPAGE analysis and by animal models. Results:The799 bp amplifiedompRgene product from wild typeS. typhiSS3 andSS5 illustrate the presence of virulent gene.Interestingly, there was only a282 bp amplified product fromS. typhiSS3 andSS5 grown in the presence of0.5,0.6 and0.7MNaCl.This illustrates the occurrence of deletion mutation inompRgene at high concentration ofNaCl.Furthermore, both the wild-type and mutantS. typhi outer membraneSDS-PAGE profile reveals the differences in the expression ofompF,ompC andompA proteins.In mice, wild type and mutant strains lethal dose (LD50) were determined.The mice died within72 h when both the wild type strains were injected intraperitoneally with3 logCFU.mL-1.When the mice were injected with the mutants in same dosage, no clinical symptoms were observed; whereas the serum antibody titre was elicited within two weeks indicated that the mutants have the ability to induce protective humoral immune response.These results suggest thatS. typhiSS3 andSS5 may be used as good candidate strains for the development of live attenuated vaccine against salmonellosis.Conclusions:This study demonstrates that theS. typhistrains were attenuated and could be good vaccine candidates in future.

  19. Draft Genome Sequence of Salmonella enterica subsp. enterica Serovar Bardo Strain CRJJGF_00099 (Phylum Gammaproteobacteria)

    Science.gov (United States)

    Gupta, Sushim K.; McMillan, Elizabeth A.; Jackson, Charlene R.; Desai, Prerak T.; Porwollik, Steffen; McClelland, Michael; Hiott, Lari M.; Humayoun, Shaheen B.

    2016-01-01

    Here, we report a 4.87-Mbp draft genome sequence of the multidrug-resistant (MDR) Salmonella enterica subsp. enterica serovar Bardo strain CRJJGF_00099, isolated from dairy cattle in 2005. PMID:27634995

  20. Draft Genome Sequence of Salmonella enterica subsp. enterica Serovar Bardo Strain CRJJGF_00099 (Phylum Gammaproteobacteria).

    Science.gov (United States)

    Gupta, Sushim K; McMillan, Elizabeth A; Jackson, Charlene R; Desai, Prerak T; Porwollik, Steffen; McClelland, Michael; Hiott, Lari M; Humayoun, Shaheen B; Frye, Jonathan G

    2016-01-01

    Here, we report a 4.87-Mbp draft genome sequence of the multidrug-resistant (MDR) Salmonella enterica subsp. enterica serovar Bardo strain CRJJGF_00099, isolated from dairy cattle in 2005. PMID:27634995

  1. Recombination and population structure in Salmonella enterica.

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    Xavier Didelot

    2011-07-01

    Full Text Available Salmonella enterica is a bacterial pathogen that causes enteric fever and gastroenteritis in humans and animals. Although its population structure was long described as clonal, based on high linkage disequilibrium between loci typed by enzyme electrophoresis, recent examination of gene sequences has revealed that recombination plays an important evolutionary role. We sequenced around 10% of the core genome of 114 isolates of enterica using a resequencing microarray. Application of two different analysis methods (Structure and ClonalFrame to our genomic data allowed us to define five clear lineages within S. enterica subspecies enterica, one of which is five times older than the other four and two thirds of the age of the whole subspecies. We show that some of these lineages display more evidence of recombination than others. We also demonstrate that some level of sexual isolation exists between the lineages, so that recombination has occurred predominantly between members of the same lineage. This pattern of recombination is compatible with expectations from the previously described ecological structuring of the enterica population as well as mechanistic barriers to recombination observed in laboratory experiments. In spite of their relatively low level of genetic differentiation, these lineages might therefore represent incipient species.

  2. Enhancement of Th1-biased protective immunity against avian influenza H9N2 virus via oral co-administration of attenuated Salmonella enterica serovar Typhimurium expressing chicken interferon-α and interleukin-18 along with an inactivated vaccine

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    Rahman Md

    2012-07-01

    Full Text Available Abstract Background Control of currently circulating re-assorted low-pathogenicity avian influenza (LPAI H9N2 is a major concern for both animal and human health. Thus, an improved LPAI H9N2 vaccination strategy is needed to induce complete immunity in chickens against LPAI H9N2 virus strains. Cytokines play a crucial role in mounting both the type and extent of an immune response generated following infection with a pathogen or after vaccination. To improve the efficacy of inactivated LPAI H9N2 vaccine, attenuated Salmonella enterica serovar Typhimurium was used for oral co-administration of chicken interferon-α (chIFN-α and chicken interleukin-18 (chIL-18 as natural immunomodulators. Results Oral co-administration of S. enterica serovar Typhimurium expressing chIFN-α and chIL-18, prior to vaccination with inactivated AI H9N2 vaccine, modulated the immune response of chickens against the vaccine antigen through enhanced humoral and Th1-biased cell-mediated immunity, compared to chickens that received single administration of S. enterica serovar Typhimurium expressing either chIFN-α or chIL-18. To further test the protective efficacy of this improved vaccination regimen, immunized chickens were intra-tracheally challenged with a high dose of LPAI H9N2 virus. Combined administration of S. enterica serovar Typhimurium expressing chIFN-α and chIL-18 showed markedly enhanced protection compared to single administration of the construct, as determined by mortality, clinical severity, and feed and water intake. This enhancement of protective immunity was further confirmed by reduced rectal shedding and replication of AIV H9N2 in different tissues of challenged chickens. Conclusions Our results indicate the value of combined administration of chIFN-α and chIL-18 using a Salmonella vaccine strain to generate an effective immunization strategy in chickens against LPAI H9N2.

  3. Whole-genome sequencing of Salmonella enterica subsp. enterica serovar Cubana strains isolated from agricultural sources

    Science.gov (United States)

    We report draft genomes of Salmonella enterica subsp. enterica Serovar Cubana strain CVM42234 isolated from chick feed in 2012 and Salmonella Cubana strain 76814 isolated from swine in 2004. The genome sizes are 4,975,046 and 4,936,251 base pairs, respectively....

  4. Complete and Closed Genome Sequences of 10 Salmonella enterica subsp. enterica Serovar Anatum Isolates from Human and Bovine Sources

    Science.gov (United States)

    Nguyen, Scott V.; Bono, James L.; Smith, Timothy P. L.; Fields, Patricia I.; Dinsmore, Blake A.; Santovenia, Monica; Kelley, Christy M.; Wang, Rong; Bosilevac, Joseph M.; Harhay, Gregory P.

    2016-01-01

    Salmonella enterica is an important pathogen transmitted by numerous vectors. Genomic comparisons of Salmonella strains from disparate hosts have the potential to further our understanding of mechanisms underlying host specificities and virulence. Here, we present the closed genome and plasmid sequences of 10 Salmonella enterica subsp. enterica serovar Anatum isolates from bovine and human sources. PMID:27257192

  5. Fitness of Salmonella enterica serovar Thompson in the Cilantro Phyllosphere

    OpenAIRE

    Maria T Brandl; Mandrell, Robert E.

    2002-01-01

    The epiphytic fitness of Salmonella enterica was assessed on cilantro plants by using a strain of S. enterica serovar Thompson that was linked to an outbreak resulting from cilantro. Salmonella serovar Thompson had the ability to colonize the surface of cilantro leaves, where it was detected by confocal laser scanning microscopy (CLSM) at high densities on the veins and in natural lesions. The population sizes of two common colonizers of plant surfaces, Pantoea agglomerans and Pseudomonas chl...

  6. Herd-level diagnosis for Salmonella enterica subsp enterica serovar Dublin infection in bovine dairy herds

    NARCIS (Netherlands)

    Veling, J.; Barkema, H.W.; Schans, van de J.; Zijderveld, van F.G.; Verhoeff, J.

    2002-01-01

    Herd-level sensitivities of bacteriological and serological methods were compared in 79 bovine dairy herds, recently infected with Salmonella enterica subsp. enterica serovar Dublin. All farms experienced clinical signs of salmonellosis for the first time and had no history of vaccination against sa

  7. Complete genome sequence of salmonella enterica subsp. enterica Serovar Thompson Strain RM6836

    Science.gov (United States)

    Salmonella enterica subsp. enterica serovar Thompson (S. Thompson) strain RM6836 was isolated from lettuce in 2002. We report the complete sequence and annotation of the genome of S. Thompson strain RM6836. This is the first reported complete genome sequence for S. Thompson and will provide a point ...

  8. Regulated delayed expression of rfc enhances the immunogenicity and protective efficacy of a heterologous antigen delivered by live attenuated Salmonella enterica vaccines.

    Science.gov (United States)

    Kong, Qingke; Liu, Qing; Jansen, Angela M; Curtiss, Roy

    2010-08-23

    The Salmonella rfc gene encodes the O-antigen polymerase. We constructed three strains in which we replaced the native rfc promoter with the arabinose-dependent araC P(BAD) promoter so that rfc expression was dependent on exogenously supplied arabinose provided during in vitro growth. The three mutant strains were designed to synthesize different amounts of Rfc by altering the ribosome-binding sequence and start codon. We examined these strains for a number of in vitro characteristics compared to an isogenic Deltarfc mutant and the wild-type parent strain. One promoter-replacement mutation, DeltaP(rfc174), yielded an optimal profile, exhibiting wild-type characteristics when grown with arabinose, and Deltarfc characteristics when grown without arabinose. In addition, when administered orally, the DeltaP(rfc174) strain was completely attenuated in for virulence in mice. The DeltaP(rfc174) mutation was introduced into attenuated Salmonella vaccine strain chi9241 (DeltapabA DeltapabB DeltaasdA) followed by introduction of an Asd(+) balanced-lethal plasmid to designed for expression of the pneumococcal surface protein PspA. Mice immunized with either chi9241 or its DeltaP(rfc174) derivative expressing pspA were protected against S. pneumoniae challenge.

  9. 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.

  10. Identification of new secreted effectors in Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Geddes, Kaoru; Worley, Micah; Niemann, George; Heffron, Fred

    2005-10-01

    A common theme in bacterial pathogenesis is the secretion of bacterial products that modify cellular functions to overcome host defenses. Gram-negative bacterial pathogens use type III secretion systems (TTSSs) to inject effector proteins into host cells. The genes encoding the structural components of the type III secretion apparatus are conserved among bacterial species and can be identified by sequence homology. In contrast, the sequences of secreted effector proteins are less conserved and are therefore difficult to identify. A strategy was developed to identify virulence factors secreted by Salmonella enterica serovar Typhimurium into the host cell cytoplasm. We constructed a transposon, which we refer to as mini-Tn5-cycler, to generate translational fusions between Salmonella chromosomal genes and a fragment of the calmodulin-dependent adenylate cyclase gene derived from Bordetella pertussis (cyaA'). In-frame fusions to bacterial proteins that are secreted into the eukaryotic cell cytoplasm were identified by high levels of cyclic AMP in infected cells. The assay was sufficiently sensitive that a single secreted fusion could be identified among several hundred that were not secreted. This approach identified three new effectors as well as seven that have been previously characterized. A deletion of one of the new effectors, steA (Salmonella translocated effector A), attenuated virulence. In addition, SteA localizes to the trans-Golgi network in both transfected and infected cells. This approach has identified new secreted effector proteins in Salmonella and will likely be useful for other organisms, even those in which genetic manipulation is more difficult.

  11. Salmonella enterica Serotype Arizonae Meningitis in a Neonate

    Directory of Open Access Journals (Sweden)

    Wubishet Lakew

    2013-01-01

    Full Text Available Typhoidal and nontyphoidal salmonella infections are common causes of gastroenteritis in the community. However, salmonella only rarely causes invasive infections like meningitis. We report a 13-day-old female neonate with signs and symptoms of meningitis whose cerebrospinal fluid (CSF culture showed Salmonella enterica serotype Arizonae that was sensitive to ceftriaxone. She presented with fever and failure to feed for 2 days. Despite prompt treatment with ampicillin, gentamicin, and ceftriaxone, she developed communicating hydrocephalus, frequent seizures, and coma that progressed to death after 2 weeks of hospitalization. Salmonella enterica serotype Arizonae is a rare cause of human infection known to leading to meningitis symptoms similar to those caused by other salmonella species. This is the first report of it as a cause of meningitis in a child under one month of age. Therefore, it should be included in the differential diagnosis of Gram-negative bacillary meningitis in immunocompromised children, neonates, and those with contacts with reptiles.

  12. Complete Genome Sequences of Two Outbreak Strains of Salmonella enterica subsp. enterica Serovar Thompson Associated with Cilantro

    OpenAIRE

    Craig T Parker; Huynh, Steven; Gorski, Lisa; Cooper, Kerry K.; Miller, William G.

    2015-01-01

    Salmonella enterica subsp. enterica serovar Thompson strains RM1984 (CADPH-99A2334) and RM1986 (CADPH-99A2345) are associated with a 1999 outbreak in contaminated cilantro. We report here the complete genome sequences and annotation of these two S. Thompson strains. These genomes are distinct and provide additional data for our understanding of S. enterica.

  13. Complete Genome Sequences of Two Outbreak Strains of Salmonella enterica subsp. enterica Serovar Thompson Associated with Cilantro.

    Science.gov (United States)

    Parker, Craig T; Huynh, Steven; Gorski, Lisa; Cooper, Kerry K; Miller, William G

    2015-01-01

    Salmonella enterica subsp. enterica serovar Thompson strains RM1984 (CADPH-99A2334) and RM1986 (CADPH-99A2345) are associated with a 1999 outbreak in contaminated cilantro. We report here the complete genome sequences and annotation of these two S. Thompson strains. These genomes are distinct and provide additional data for our understanding of S. enterica. PMID:26586897

  14. Automated 5 ' nuclease PCR assay for identification of Salmonella enterica

    DEFF Research Database (Denmark)

    Hoorfar, Jeffrey; Ahrens, Peter; Rådström, P.

    2000-01-01

    A simple and ready-to-go test based on a 5' nuclease (TaqMan) PCR technique was developed for identification of presumptive Salmonella enterica isolates. The results were compared with those of conventional methods. The TaqMan assay was evaluated for its ability to accurately detect 210 S. enterica...... isolates, including 100 problematic "rough" isolates. An internal positive control was designed to use the same Salmonella primers for amplification of a spiked nonrelevant template (116 bp) in the sample tube. The PCR test correctly identified all the Salmonella strains by resulting in positive end...... Salmonella strains tested resulted in positive FAM and TET signals. In addition, it was found that the complete PCR mixture, predispensed in microwell plates, could be stored for up to 3 months at -20 degrees C, Thus, the diagnostic TaqMan assay developed can be a useful and simple alternative method...

  15. Effects of crp deletion in Salmonella enterica serotype Gallinarum

    Directory of Open Access Journals (Sweden)

    Rubino Salvatore

    2007-05-01

    Full Text Available Abstract Background Salmonella enterica serotype Gallinarum (S. Gallinarum remains an important pathogen of poultry, especially in developing countries. There is a need to develop effective and safe vaccines. In the current study, the effect of crp deletion was investigated with respect to virulence and biochemical properties and the possible use of a deletion mutant as vaccine candidate was preliminarily tested. Methods Mutants were constructed in S. Gallinarum by P22 transduction from Salmonella Typhimurium (S. Typhimurium with deletion of the crp gene. The effect was characterized by measuring biochemical properties and by testing of invasion in a chicken loop model and by challenge of six-day-old chickens. Further, birds were immunized with the deleted strain and challenged with the wild type isolate. Results The crp deletions caused complete attenuation of S. Gallinarum. This was shown by ileal loop experiments not to be due to significantly reduced invasion. Strains with such deletions may have vaccine potential, since oral inoculatoin with S. Gallinarum Δcrp completely protected against challenge with the same dose of wild type S. Gallinarum ten days post immunization. Interestingly, the mutations did not cause the same biochemical and growth changes to the two biotypes of S. Gallinarum. All biochemical effects but not virulence could be complemented by providing an intact crp-gene from S. Typhimurium on the plasmid pSD110. Conclusion Transduction of a Tn10 disrupted crp gene from S. Typhimurium caused attenuation in S. Gallinarum and mutated strains are possible candidates for live vaccines against fowl typhoid.

  16. Analysis of the contribution of bacteriophage ST64B to in vitro virulence traits of Salmonella enterica serovar Typhimurium

    DEFF Research Database (Denmark)

    Fresno, Ana Herrero; Leekitcharoenphon, Pimlapas; Hendriksen, Rene S.;

    2014-01-01

    Comparison of the publicly available genomes of the virulent Salmonella enterica serovar Typhimurium (S. Typhimurium) strains SL1344, 14028s and D23580 to that of the virulence-attenuated isolate LT2 revealed the absence of a full sequence of bacteriophage ST64B in the latter. Four selected ST64B...

  17. Ceftriaxone-resistant Salmonella enterica serotype Newport, France

    OpenAIRE

    Egorova, S.; Timinouni, M.; Demartin, M.; Granier, S.; Whichard, J.; Sangal, V; Fabre, L; Delaune, A.; Pardos, M.; Millemann, Y.; Espie, E; Achtman, M; Grimont, P; Weill, F.

    2008-01-01

    The multidrug-resistant (MDR) Salmonella enterica serotype Newport strain that produces CMY-2 β-lactamase(Newport MDR-AmpC) was the source of sporadic cases and outbreaks in humans in France during 2000–2005. Because this strain was not detected in food animals, it was most likely introduced into France through imported food products.

  18. Ceftriaxone-Resistant Salmonella enterica Serotype Newport, France

    OpenAIRE

    Egorova, Svetlana; Timinouni, Mohammed; Demartin, Marie; Granier, Sophie; Whichard, Jean; Sangal, Vartul; Fabre, Laëtitia; Delauné, Aurélia; Pardos, Maria; Millemann, Yves; Espié, Emmanuelle; Achtman, Mark; Grimont, Patrick; Weill, François-Xavier

    2008-01-01

    The multidrug-resistant (MDR) Salmonella enterica serotype Newport strain that produces CMY-2 beta-lactamase (Newport MDR-AmpC) was the source of sporadic cases and outbreaks in humans in France during 2000-2005. Because this strain was not detected in food animals, it was most likely introduced into France through imported food products.

  19. lac repressor is an antivirulence factor of Salmonella enterica: its role in the evolution of virulence in Salmonella.

    Directory of Open Access Journals (Sweden)

    Sandeepa M Eswarappa

    Full Text Available The genus Salmonella includes many pathogens of great medical and veterinary importance. Bacteria belonging to this genus are very closely related to those belonging to the genus Escherichia. lacZYA operon and lacI are present in Escherichia coli, but not in Salmonella enterica. It has been proposed that Salmonella has lost lacZYA operon and lacI during evolution. In this study, we have investigated the physiological and evolutionary significance of the absence of lacI in Salmonella enterica. Using murine model of typhoid fever, we show that the expression of LacI causes a remarkable reduction in the virulence of Salmonella enterica. LacI also suppresses the ability of Salmonella enterica to proliferate inside murine macrophages. Microarray analysis revealed that LacI interferes with the expression of virulence genes of Salmonella pathogenicity island 2. This effect was confirmed by RT-PCR and Western blot analysis. Interestingly, we found that SBG0326 of Salmonella bongori is homologous to lacI of Escherichia coli. Salmonella bongori is the only other species of the genus Salmonella and it lacks the virulence genes of Salmonella pathogenicity island 2. Overall, our results demonstrate that LacI is an antivirulence factor of Salmonella enterica and suggest that absence of lacI has facilitated the acquisition of virulence genes of Salmonella pathogenicity island 2 in Salmonella enterica making it a successful systemic pathogen.

  20. Salmonella enterica induces and subverts the plant immune system

    KAUST Repository

    García, Ana V.

    2014-04-04

    Infections with Salmonella enterica belong to the most prominent causes of food poisoning and infected fruits and vegetables represent important vectors for salmonellosis. Although it was shown that plants raise defense responses against Salmonella, these bacteria persist and proliferate in various plant tissues. Recent reports shed light into the molecular interaction between plants and Salmonella, highlighting the defense pathways induced and the means used by the bacteria to escape the plant immune system and accomplish colonization. It was recently shown that plants detect Salmonella pathogen-associated molecular patterns (PAMPs), such as the flagellin peptide flg22, and activate hallmarks of the defense program known as PAMP-triggered immunity (PTI). Interestingly, certain Salmonella strains carry mutations in the flg22 domain triggering PTI, suggesting that a strategy of Salmonella is to escape plant detection by mutating PAMP motifs. Another strategy may rely on the type III secretion system (T3SS) as T3SS mutants were found to induce stronger plant defense responses than wild type bacteria. Although Salmonella effector delivery into plant cells has not been shown, expression of Salmonella effectors in plant tissues shows that these bacteria also possess powerful means to manipulate the plant immune system. Altogether, these data suggest that Salmonella triggers PTI in plants and evolved strategies to avoid or subvert plant immunity. 2014 Garca and Hirt.

  1. Resistance to antibiotics and genotype characteristics of Salmonella enterica subspecies enterica serovar Mbandaka isolated from poultry

    Directory of Open Access Journals (Sweden)

    Vidanović Dejan

    2008-01-01

    Full Text Available Salmonellas are one of the main zoonotic pathogens whose reservoirs are poultry, cattle and pigs. By means of the food chain salmonellas can be transferred to humans through contaminated food of animal origin. Multiresistant strains Salmonella are particularly dangerous since they can transfer genes of resistance to antibiotics to other microorganisms. Control of salmonellas primarily depends on a good surveillance system and knowledge of the strain types present in the epizootiologic area. In some geographical regions only a few Salmonella serotypes are usually of epidemiological importance. Due to the predomination of some serotypes and fagotypes, when an additional discrimination within serotypes and fagotypes is needed, DNA genotyping is used. In cases when it is necessary to compare the strains which caused the poisoning of patients, with strains isolated from food or animals, a highly discriminatory method is used - pulsed field gel electrophoresis (PFGE. Due to a high degree of discrimination the results of PFGE testing enable decision making with a higher degree of certainty in epizootiologic and epidemiologic research work. The aim of this testing was to determine the antibiotics resistance and genotype characteristics of Salmonella enterica subsp. enterica serovar Mbandaka isolated from poultry from some areas of the territory of Serbia.

  2. FAKTOR VIRULENSI Salmonella enterica SEROVAR TYPHI

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    Marvy Khrisna Pranamartha

    2015-09-01

    Full Text Available ABSTRAK Demam tifoid disebabkan oleh bakteri Salmonella typhi, dengan gejala umum berupa demam tinggi dan nyeri perut. Tifoid adalah penyakit infeksi yang disebabkan oleh bakteri Salmonella typhi, yang masuk ke dalam tubuh melalui mulut dan saluran cerna.1 Untuk bisa memahami patogenesis dari demam tifoid sampai ke tingkat selular dan molekular, ada 5 hal penting yang harus digaris bawahi, yaitu: 1.\tTipe 3 Sistem Sekresi (T3SS 2.\tVirulence Genes dari Salmonella yang mengkode 5 SIP (Salmonella Invasion Protein SIP A, B, C, D, dan E. 3.\tToll R2 dan toll R3 yang merupakan lapisan luar dari makrofag. 4.\tSistem imun lumen usus sampai ke organ dalam 5.\tFungsi endotelial sel dalam inflamasi. Infeksi Salmonella dapat berakibat fatal kepada bayi, balita, ibu hamil dan kandungannya serta orang lanjut usia. Hal ini disebabkan karena kekebalan tubuh mereka yang menurun. Virulensi salmonella tidak lepas dari peranan SPI, yang terletak di dalam kromosom dan plasmid bakteri. Dimana SPI 1 dan SPI 2 telah dikaji cukup mendalam karena keterkaitannya dengan T3SS, dan berperan sangat penting pada invasi awal serta siklus hidup intrasel dari bakteri Salmonella. Kontaminasi Salmonella dapat dicegah dengan mencuci tangan dan menjaga kebersihan makanan yang dikonsumsi. Selalu menjaga kebersihan lingkungan hidup kita agar terhindar dari kontaminasi dengan bakteri Salmonella typhi. Agar mewaspadai sejak dini pencegahan dan pengobatan penyakit typhus. Studi mendalam perlu dilakukan agar kita mampu lebih memahami proses kompleks antara patogen dan sel inang. Mengingat dari 15 SPI yang sudah diketahui, hanya SPI 1 dan SPI 2 yang sudah dikaji secara mendalam. Kata Kunci: Salmonella, Salmonella Invasion Protein, Typhi.

  3. Polyamines Are Required for Virulence in Salmonella enterica Serovar Typhimurium

    DEFF Research Database (Denmark)

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

    2012-01-01

    Sensing and responding to environmental cues is a fundamental characteristic of bacterial physiology and virulence. Here we identify polyamines as novel environmental signals essential for virulence of Salmonella enterica serovar Typhimurium, a major intracellular pathogen and a model organism......, 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...

  4. Increasing quinolone resistance in Salmonella enterica serotype enteritidis

    DEFF Research Database (Denmark)

    Mølbak, K.; Gerner-Smidt, P.; Wegener, Henrik Caspar

    2002-01-01

    Until recently, Salmonella enterica serotype Enteritidis has remained sensitive to most antibiotics. However, national surveillance data from Denmark show that quinolone resistance in S. Enteritidis has increased from 0.8% in 1995 to 8.5% in 2000. These data support concerns that the current use of...... quinolone in food animals leads to increasing resistance in S. Enteritidis and that action should be taken to limit such use....

  5. Complete genomic sequences of two outbreak strains of Salmonella enterica subsp. enterica serovar Thompson associated with cilantro

    Science.gov (United States)

    Salmonella enterica subsp. enterica serovar Thompson strains RM1984 (CADPH-99A2334) and RM1986 (CADPH -99A2345) are clinical isolates from 1999, putatively related to an outbreak in California from contaminated cilantro. We report the complete genome sequences and annotation of these two S. Thompson...

  6. Complete Genome Sequences of 17 Canadian Isolates of Salmonella enterica subsp. enterica Serovar Heidelberg from Human, Animal, and Food Sources.

    Science.gov (United States)

    Labbé, Geneviève; Ziebell, Kim; Bekal, Sadjia; Macdonald, Kimberley A; Parmley, E Jane; Agunos, Agnes; Desruisseau, Andrea; Daignault, Danielle; Slavic, Durda; Hoang, Linda; Ramsay, Danielle; Pollari, Frank; Robertson, James; Nash, John H E; Johnson, Roger P

    2016-01-01

    Salmonella enterica subsp. enterica serovar Heidelberg is a highly clonal serovar frequently associated with foodborne illness. To facilitate subtyping efforts, we report fully assembled genome sequences of 17 Canadian S Heidelberg isolates including six pairs of epidemiologically related strains. The plasmid sequences of eight isolates contain several drug resistance genes.

  7. Complete Genome Sequences of 17 Canadian Isolates of Salmonella enterica subsp. enterica Serovar Heidelberg from Human, Animal, and Food Sources

    Science.gov (United States)

    Labbé, Geneviève; Ziebell, Kim; Bekal, Sadjia; Parmley, E. Jane; Agunos, Agnes; Desruisseau, Andrea; Daignault, Danielle; Slavic, Durda; Hoang, Linda; Ramsay, Danielle; Pollari, Frank; Robertson, James; Nash, John H. E.

    2016-01-01

    Salmonella enterica subsp. enterica serovar Heidelberg is a highly clonal serovar frequently associated with foodborne illness. To facilitate subtyping efforts, we report fully assembled genome sequences of 17 Canadian S. Heidelberg isolates including six pairs of epidemiologically related strains. The plasmid sequences of eight isolates contain several drug resistance genes. PMID:27635008

  8. Complete Genome Sequences of 17 Canadian Isolates of Salmonella enterica subsp. enterica Serovar Heidelberg from Human, Animal, and Food Sources.

    Science.gov (United States)

    Labbé, Geneviève; Ziebell, Kim; Bekal, Sadjia; Macdonald, Kimberley A; Parmley, E Jane; Agunos, Agnes; Desruisseau, Andrea; Daignault, Danielle; Slavic, Durda; Hoang, Linda; Ramsay, Danielle; Pollari, Frank; Robertson, James; Nash, John H E; Johnson, Roger P

    2016-01-01

    Salmonella enterica subsp. enterica serovar Heidelberg is a highly clonal serovar frequently associated with foodborne illness. To facilitate subtyping efforts, we report fully assembled genome sequences of 17 Canadian S Heidelberg isolates including six pairs of epidemiologically related strains. The plasmid sequences of eight isolates contain several drug resistance genes. PMID:27635008

  9. Evaluation of Salmonella enterica type III secretion system effector proteins as carriers for heterologous vaccine antigens.

    Science.gov (United States)

    Hegazy, Wael Abdel Halim; Xu, Xin; Metelitsa, Leonid; Hensel, Michael

    2012-03-01

    Live attenuated strains of Salmonella enterica have a high potential as carriers of recombinant vaccines. The type III secretion system (T3SS)-dependent translocation of S. enterica can be deployed for delivery of heterologous antigens to antigen-presenting cells. Here we investigated the efficacy of various effector proteins of the Salmonella pathogenicity island (SPI2)-encoded T3SS for the translocation of model antigens and elicitation of immune responses. The SPI2 T3SS effector proteins SifA, SteC, SseL, SseJ, and SseF share an endosomal membrane-associated subcellular localization after translocation. We observed that all effector proteins could be used to translocate fusion proteins with the model antigens ovalbumin and listeriolysin into the cytosol of host cells. Under in vitro conditions, fusion proteins with SseJ and SteC stimulated T-cell responses that were superior to those triggered by fusion proteins with SseF. However, in mice vaccinated with Salmonella carrier strains, only fusion proteins based on SseJ or SifA elicited potent T-cell responses. These data demonstrate that the selection of an optimal SPI2 effector protein for T3SS-mediated translocation is a critical parameter for the rational design of effective Salmonella-based recombinant vaccines.

  10. Immunogenicity of Salmonella enterica serovar Enteritidis virulence protein, InvH, and cross-reactivity of its antisera with Salmonella strains.

    Science.gov (United States)

    Dehghani, Behzad; Rasooli, Iraj; Gargari, Seyed Latif Mousavi; Nadooshan, Mohammad Reza Jalali; Owlia, Parviz; Nazarian, Shahram

    2013-02-22

    Acellular vaccines containing bacterial immunodominant components such as surface proteins may be potent alternatives to live attenuated vaccines in order to reduce salmonellosis risk to human health. invH gene, an important part of needle complex in type three secretion system (TTSS) plays important role in efficient bacterial adherence and entry into epithelial cells. In this work we hypothesize that use of a 15 kDa recombinant InvH as Salmonella enterica serovar Enteritidis surface protein could provoke antibody production in mouse and would help us study feasibility of its potential for diagnosis and/or a recombinant vaccine. The purified InvH provoked significant rise of IgG in mice. Active protection induced by immunization with InvH against variable doses of S. enterica serovar Enteritidis, indicated that the immunized mice were completely protected against challenge with 10(4) LD(50). The immunoreaction of sera from immunized mice with other Salmonella strains or cross reaction with sera of Salmonella strains inoculated mice is indicative of possessing by Salmonella strains of the surface protein, InvH, that can be employed in both prophylactic and diagnostic measures against S. enterica. Bacteria free spleen and ileum of the immunized mice in this study indicate that the invH gene affects bacterial invasion. Efficacy of the virulence protein, InvH, in shuttling into host cells in injectisome of S. enterica serovar Enteritidis and inhibition of this phenomenon by active immunization was shown in this study. In conclusion immunization with InvH protein can develop protection against S. enterica serovar Enteritidis infections. InvH in Salmonella strains can be exploited in protective measures as well as a diagnostic tool in Salmonella infections.

  11. Repeated isolation of Salmonella enterica Goverdhan, a very rare serovar, from Danish poultry surveillance samples

    DEFF Research Database (Denmark)

    Pedersen, Karl; Sørensen, Gitte; Szabo, Istvan;

    2014-01-01

    We report here the appearance of a very rare serovar of Salmonella, S. enterica subsp. enterica serovar Goverdhan, in routine Salmonella surveillance samples from Danish poultry production. S. Goverdhan was found on nine occasions: in one broiler breeder farm in October 2010, four broiler farms a...

  12. Real-time monitoring of Salmonella enterica in free-range geese

    DEFF Research Database (Denmark)

    Christensen, Laurids Siig; Josefsen, Mathilde Hartmann; Pedersen, Karl;

    2011-01-01

    Free-range geese were sampled longitudinally and Salmonella isolates characterized to reveal highly diverging colonization dynamics. One flock was intermittently colonized with one strain of Salmonella enterica serovar Enteritidis from 2 weeks of age, while in another, S. enterica serovar Mbandaka...

  13. Uncovering what lies beneath a Salmonella enterica empyema.

    Science.gov (United States)

    Woo, Jia Wei; Tam, John Kit Chung; Chan, Douglas Su Gin; Wang, Shi; Ying, Lee Shir

    2015-01-01

    A 67-year-old woman with myelodysplastic syndrome (MDS) and transfusional haemosiderosis developed Salmonella empyema caused by direct extension from splenic abscesses. She was successfully treated with antibiotics, pleural decortication and splenectomy. She had presented with fever after being treated for presumed pneumonia and parapneumonic effusion 2 months prior. CT scan showed splenic abscesses eroding through the diaphragm causing a left pleural empyema. Pleural fluid and spleen bacterial cultures grew Salmonella enterica. She was treated with 4 weeks of antibiotics and underwent surgical pleural decortication and splenectomy in the same sitting. She made a good postoperative recovery. Patients with severe iron overload are susceptible to various types of bacterial sepsis, including salmonellosis. It is unusual for enteric bacterial such as Salmonella to present with empyema, and should prompt a search for intra-abdominal infection. Pleural decortication and splenectomy can be performed during the same surgical sitting and can lead to good surgical outcomes. PMID:26336186

  14. 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.

  15. Quinolone Resistance Mechanisms Among Salmonella enterica in Malaysia.

    Science.gov (United States)

    Thong, Kwai Lin; Ngoi, Soo Tein; Chai, Lay Ching; Teh, Cindy Shuan Ju

    2016-06-01

    The prevalence of quinolone-resistant Salmonella enterica is on the rise worldwide. Salmonella enterica is one of the major foodborne pathogens in Malaysia. Therefore, we aim to investigate the occurrence and mechanisms of quinolone resistance among Salmonella strains isolated in Malaysia. A total of 283 Salmonella strains isolated from food, humans, and animals were studied. The disk diffusion method was used to examine the quinolone susceptibility of the strains, and the minimum inhibitory concentration (MIC) values of nalidixic acid and ciprofloxacin were also determined. DNA sequencing of the quinolone resistance-determining regions (QRDRs) of gyrase and topoisomerase IV genes and the plasmid-borne qnr genes was performed. The transfer of the qnr gene was examined through transconjugation experiment. A total of 101 nalidixic acid-resistant Salmonella strains were identified. In general, all strains were highly resistant to nalidixic acid (average MICNAL, 170 μg/ml). Resistance to ciprofloxacin was observed in 30.7% of the strains (1 ≤ MICCIP ≤ 2 μg/ml). Majority of the strains contained missense mutations in the QRDR of gyrA (69.3%). Silent mutations were frequently detected in gyrB (75.2%), parC (27.7%), and parE (51.5%) within and beyond the QRDRs. Novel mutations were detected in parC and parE. The plasmid-borne qnrS1 variant was found in 36.6% of the strains, and two strains were found to be able to transfer the qnrS1 gene. Overall, mutations in gyrA and the presence of qnrS1 genes might have contributed to the high level of quinolone resistance among the strains. Our study provided a better understanding on the status of quinolone resistance among Salmonella strains circulating in Malaysia. PMID:26683630

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

    OpenAIRE

    Mattick, K. L.; Jørgensen, F.; Legan, J. D.; Cole, M. B.; Porter, J.(Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA); 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 NaC...

  17. Changing trends in antimicrobial resistance of Salmonella enterica serovar typhi and salmonella enterica serovar paratyphi A in Chennai

    Directory of Open Access Journals (Sweden)

    Krishnan Padma

    2009-10-01

    Full Text Available Background and Objectives: Chloramphenicol was considered the anti-microbial gold standard for typhoid treatment but, following the increasing worldwide frequency of antibiotic resistance, ciprofloxacin has been the mainstay of therapy since 1980. Recent studies have shown a shifting of susceptibility to conventional drugs like chloramphenicol, ampicillin and cotrimoxazole. The primary objective of the study was to evaluate the in vitro activity of chloramphenicol and other first-line drugs in comparison with cephalosporins and quinolones. Materials and Methods: Fifty isolates of Salmonella obtained from blood culture were subjected to serotyping at the Central Research Institute, Kasauli. Phage typing and biotyping was performed at the National Phage Typing Centre, New Delhi. Antibiotic sensitivity testing was carried out for 10 drugs by the Kirby-Bauer disc diffusion method and minimum inhibitory concentration by broth microdilution for nalidixic acid, chloramphenicol, ciprofloxacin, ceftriaxone, cefixime and ofloxacin. Multi-drug-resistant (MDR strains were checked for plasmid. Results: In the present study, 70 and 30% of the isolates were Salmonella enterica serovar typhi and paratyphi A, respectively. They were highly sensitive to chloramphenicol (86%, ampicillin (84% and cotrimoxazole (88%. Highest sensitivity was seen for cephalosporins, followed by quinolones. Seventeen/21 (81% and 100% of the Salmonella enterica serovar typhi strains belonged to E1 phage type and biotype 1, respectively. Antibiogram showed 2% of the strains to be sensitive to all the drugs tested and 12% were MDR and showed the presence of plasmids. Conclusion: The study indicates reemergence of chloramphenicol-susceptible Salmonella enterica serovar typhi and paratyphi A isolates, a significant decline in MDR strains and high resistance to nalidixic acid. E1 phage type and biotype 1 are found to be most prevalent in Chennai, India.

  18. Genome-wide methylation patterns in Salmonella enterica Subsp. enterica Serovars.

    Directory of Open Access Journals (Sweden)

    Cary Pirone-Davies

    Full Text Available The methylation of DNA bases plays an important role in numerous biological processes including development, gene expression, and DNA replication. Salmonella is an important foodborne pathogen, and methylation in Salmonella is implicated in virulence. Using single molecule real-time (SMRT DNA-sequencing, we sequenced and assembled the complete genomes of eleven Salmonella enterica isolates from nine different serovars, and analysed the whole-genome methylation patterns of each genome. We describe 16 distinct N6-methyladenine (m6A methylated motifs, one N4-methylcytosine (m4C motif, and one combined m6A-m4C motif. Eight of these motifs are novel, i.e., they have not been previously described. We also identified the methyltransferases (MTases associated with 13 of the motifs. Some motifs are conserved across all Salmonella serovars tested, while others were found only in a subset of serovars. Eight of the nine serovars contained a unique methylated motif that was not found in any other serovar (most of these motifs were part of Type I restriction modification systems, indicating the high diversity of methylation patterns present in Salmonella.

  19. Isolation and characterization of Salmonella enterica Weltevreden cytotoxin.

    Science.gov (United States)

    Singh, Y; Sharma, V D

    1998-03-01

    Cytotoxin of Salmonella enterica subspecies enterica serovar Weltevreden (BM-1643), isolated from buffalo meat, was purified and characterized physicochemically and immunologically. Cell-free culture supernatant (CFCS) of the organism showing marked cytotoxicity to Vero cells and least enterotoxicity to rabbit ligated ileal loop (RLIL) model, was salt precipitated with ammonium sulphate (60% saturation level) and dialysed. Precipitated dialysed preparation (60% PDP) when filtered through Sephadex G-100 column yielded two peaks, of which second peak (SG-100 SP) contained the cytotoxic activity. Upon filtration of SG-100 SP through SG-200 column, three peaks were obtained. Second peak (SG-200 SP), which was cytotoxic, yielded a single protein band of approximately 60-70 kDa in polyacrylamide gel electrophoresis and 3 protein bands of lower, molecular weight (13.5-56 kDa) in SDS-PAGE analysis. Cytotoxic preparation was maximally active at pH 7 to 8. On heating above 60 degrees C, cytotoxicity decreased gradually with insignificant activity left after treatment at 121 degrees C (15 min). Cytotoxin was inactivated by treatment with trypsin and protease but not by papain or lipase enzymes. It was immunogenic in rabbit and antiserum neutralized the cytotoxicity of cytotoxic preparations of homologous as well as heterologous Salmonella serovars. PMID:9754063

  20. Virulent Salmonella enterica serovar typhimurium evades adaptive immunity by preventing dendritic cells from activating T cells.

    Science.gov (United States)

    Tobar, Jaime A; Carreño, Leandro J; Bueno, Susan M; González, Pablo A; Mora, Jorge E; Quezada, Sergio A; Kalergis, Alexis M

    2006-11-01

    Dendritic cells (DCs) constitute the link between innate and adaptive immunity by directly recognizing pathogen-associated molecular patterns (PAMPs) in bacteria and by presenting bacterial antigens to T cells. Recognition of PAMPs renders DCs as professional antigen-presenting cells able to prime naïve T cells and initiate adaptive immunity against bacteria. Therefore, interfering with DC function would promote bacterial survival and dissemination. Understanding the molecular mechanisms that have evolved in virulent bacteria to evade activation of adaptive immunity requires the characterization of virulence factors that interfere with DC function. Salmonella enterica serovar Typhimurium, the causative agent of typhoid-like disease in the mouse, can prevent antigen presentation to T cells by avoiding lysosomal degradation in DCs. Here, we show that this feature of virulent Salmonella applies in vivo to prevent activation of adaptive immunity. In addition, this attribute of virulent Salmonella requires functional expression of a type three secretion system (TTSS) and effector proteins encoded within the Salmonella pathogenicity island 2 (SPI-2). In contrast to wild-type virulent Salmonella, mutant strains carrying specific deletions of SPI-2 genes encoding TTSS components or effectors proteins are targeted to lysosomes and are no longer able to prevent DCs from activating T cells in vitro or in vivo. SPI-2 mutant strains are attenuated in vivo, showing reduced tissue colonization and enhanced T-cell activation, which confers protection against a challenge with wild-type virulent Salmonella. Our data suggest that impairment of DC function by the activity of SPI-2 gene products is crucial for Salmonella pathogenesis.

  1. Complete genome sequence of a multiple drug resistant Salmonella enterica serovar Typhi CT18

    DEFF Research Database (Denmark)

    Parkhill, J.; Dougan, G.; James, K.D.;

    2001-01-01

    Salmonella enterica serovar Typhi (S. typhi) is the aetiological agent of typhoid fever, a serious invasive bacterial disease of humans with an annual global burden of approximately 16 million cases, leading to 600,000 fatalities(1). Many S. enterica serovars actively invade the mucosal surface o...

  2. Clustered Intracellular Salmonella enterica Serovar Typhimurium Blocks Host Cell Cytokinesis.

    Science.gov (United States)

    Santos, António J M; Durkin, Charlotte H; Helaine, Sophie; Boucrot, Emmanuel; Holden, David W

    2016-07-01

    Several bacterial pathogens and viruses interfere with the cell cycle of their host cells to enhance virulence. This is especially apparent in bacteria that colonize the gut epithelium, where inhibition of the cell cycle of infected cells enhances the intestinal colonization. We found that intracellular Salmonella enterica serovar Typhimurium induced the binucleation of a large proportion of epithelial cells by 14 h postinvasion and that the effect was dependent on an intact Salmonella pathogenicity island 2 (SPI-2) type 3 secretion system. The SPI-2 effectors SseF and SseG were required to induce binucleation. SseF and SseG are known to maintain microcolonies of Salmonella-containing vacuoles close to the microtubule organizing center of infected epithelial cells. During host cell division, these clustered microcolonies prevented the correct localization of members of the chromosomal passenger complex and mitotic kinesin-like protein 1 and consequently prevented cytokinesis. Tetraploidy, arising from a cytokinesis defect, is known to have a deleterious effect on subsequent cell divisions, resulting in either chromosomal instabilities or cell cycle arrest. In infected mice, proliferation of small intestinal epithelial cells was compromised in an SseF/SseG-dependent manner, suggesting that cytokinesis failure caused by S Typhimurium delays epithelial cell turnover in the intestine.

  3. Draft Genome Sequences of 33 Salmonella enterica Clinical and Wildlife Isolates from Chile

    OpenAIRE

    Toro, Magaly; Retamal, Patricio; Allard, Marc; Brown, Eric W.; Evans, Peter; Gonzalez-Escalona, Narjol

    2015-01-01

    Salmonella enterica causes health problem worldwide. The relationships among strains that are from the same serotype but different hosts, countries, and continents remain elusive. Few genome sequences are available from S. enterica isolates from South America. Therefore, we sequenced the genomes of 33 strains from diverse sources isolated in Chile and determined that they were of different serotypes. These genomes will improve phylogenetic analysis of Salmonella strains from Chile and the res...

  4. Salmonella enterica Serotype Bredeney: Antimicrobial Susceptibility and Molecular Diversity of Isolates from Ireland and Northern Ireland

    OpenAIRE

    Cormican, Martin; DeLappe, Niall; O’Hare, Colette; Doran, Geraldine; Morris, Dearbhaile; Corbett-Feeney, Geraldine; Fanning, Séamus; Daly, Mairead; Fitzgerald, Margaret; Moore, John

    2002-01-01

    Salmonella enterica serotype Bredeney has emerged as the third most commonly identified serotype among human clinical isolates referred to the Irish National Salmonella Reference Laboratory in the years 1998 to 2000. A collection of 112 isolates of S. enterica serotype Bredeney collected during the period 1995 to 1999 from animal, food, and human sources from both Ireland and Northern Ireland were studied. Antimicrobial susceptibility testing, pulsed-field gel electrophoresis (PFGE), and DNA ...

  5. RECOVERY IN VIVO OF NONCULTURABLE SUBPOPULATION OF SALMONELLA ENTERICA

    Directory of Open Access Journals (Sweden)

    Yudin I.P.

    2015-12-01

    Full Text Available Introduction. As one of mesophilic, easily cultivated species of pathogenic bacteria, Salmonella enterica transformed into viable but nonculturable (VNC state in response to environmental stresses, including action of biocides. The cells in this state, preserve the integrity of membranes and metabolism of some, but not detected by conventional methods of cultivation. Some researchers suggest that the evolutionary significance of this phenomenon is part of an adaptive response aimed at long-term survival of bacteria in adverse conditions; others argue that it is the result of stochastic cellular damage, in which nonculturable cells are in a state of gradual death. In any case, the phenomenon of existence VNC pathogens if they retain the ability to restore its growth in vivo is a significant problem in medicine, pharmaceutical, veterinary, food industry. VNC subpopulation of S. enterica was obtained under action of ethanol. In this paper was investigated in vivo resuscitation VNC S. enterica using intraperitoneal injection of mice. Materials and methods. Obtaining of stressful S. enterica populations. Bacteria were grown to exponential phase in broth Luria–Bertani (LB. To 1.0 ml sample suspension diluted to 1.5 × 106 cells/ml was added 1.0 ml of ethanol at a concentration of 40 % (v/v. After exposure of 10 to 600 minutes in the suspension were added 8.0 ml of phosphate buffered saline (FBS, washed by centrifugation (4500 g for 5 minutes and serially diluted at a ratio of 1:10 (v/v samples were stained with LIVE/DEAD BacLight (produced by "Invitrogen", USA, filtrated on membrane filters for fluorescence microscopy and parallel plated on LB agar cup to determine colony-forming units (CFU per ml. In vivo resuscitation VNC S. enterica was made following way. Three groups of animals were inoculated by intraperitoneal injection: 1 103 culturable cells (0.1 ml suspension containing 104 CFU / ml; 2 103 VNC cells (0.1 ml suspension containing 104 cells

  6. The ferric enterobactin transporter Fep is required for persistent Salmonella enterica serovar typhimurium infection.

    Science.gov (United States)

    Nagy, Toni A; Moreland, Sarah M; Andrews-Polymenis, Helene; Detweiler, Corrella S

    2013-11-01

    Most bacterial pathogens require iron to grow and colonize host tissues. The Gram-negative bacterium Salmonella enterica serovar Typhimurium causes a natural systemic infection of mice that models acute and chronic human typhoid fever. S. Typhimurium resides in tissues within cells of the monocyte lineage, which limit pathogen access to iron, a mechanism of nutritional immunity. The primary ferric iron import system encoded by Salmonella is the siderophore ABC transporter FepBDGC. The Fep system has a known role in acute infection, but it is unclear whether ferric iron uptake or the ferric iron binding siderophores enterobactin and salmochelin are required for persistent infection. We defined the role of the Fep iron transporter and siderophores in the replication of Salmonella in macrophages and in mice that develop acute followed by persistent infections. Replication of wild-type and iron transporter mutant Salmonella strains was quantified in cultured macrophages, fecal pellets, and host tissues in mixed- and single-infection experiments. We show that deletion of fepB attenuated Salmonella replication and colonization within macrophages and mice. Additionally, the genes required to produce and transport enterobactin and salmochelin across the outer membrane receptors, fepA and iroN, are needed for colonization of all tissues examined. However, salmochelin appears to be more important than enterobactin in the colonization of the spleen and liver, both sites of dissemination. Thus, the FepBDGC ferric iron transporter and the siderophores enterobactin and salmochelin are required by Salmonella to evade nutritional immunity in macrophages and cause persistent infection in mice.

  7. Biological and virulence characteristics of Salmonella enterica serovar Typhimurium following deletion of glucose-inhibited division (gidA) gene.

    Science.gov (United States)

    Shippy, Daniel C; Eakley, Nicholas M; Bochsler, Philip N; Chopra, Ashok K; Fadl, Amin A

    2011-06-01

    Salmonella enterica serovar Typhimurium is a frequent cause of enteric disease due to the consumption of contaminated food. Identification and characterization of bacterial factors involved in Salmonella pathogenesis would help develop effective strategies for controlling salmonellosis. To investigate the role of glucose-inhibited division gene (gidA) in Salmonella virulence, we constructed a Salmonella mutant strain in which gidA was deleted. Deletion of gidA rendered Salmonella deficient in the invasion of intestinal epithelial cells, bacterial motility, intracellular survival, and induction of cytotoxicity in host cells. Deletion of gidA rendered the organism to display a filamentous morphology compared to the normal rod-shaped nature of Salmonella. Furthermore, a significant attenuation in the induction of inflammatory cytokines and chemokines, histopathological lesions, and systemic infection was observed in mice infected with the gidA mutant. Most importantly, a significant increase in LD(50) was observed in mice infected with the gidA mutant, and mice immunized with the gidA mutant were able to survive a lethal dose of wild-type Salmonella. Additionally, deletion of gidA significantly altered the expression of several bacterial factors associated with pathogenesis as indicated by global transcriptional and proteomic profiling. Taken together, our data indicate GidA as a potential regulator of Salmonella virulence genes.

  8. Regulation of biofilm formation in Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Simm, Roger; Ahmad, Irfan; Rhen, Mikael; Le Guyon, Soazig; Römling, Ute

    2014-01-01

    In animals, plants and the environment, Salmonella enterica serovar Typhimurium forms the red dry and rough (rdar) biofilm characterized by extracellular matrix components curli and cellulose. With complex expression control by at least ten transcription factors, the bistably expressed orphan response regulator CsgD directs rdar morphotype development. CsgD expression is an integral part of the Hfq regulon and the complex cyclic diguanosine monophosphate signaling network partially controlled by the global RNA-binding protein CsrA. Cell wall turnover and the periplasmic redox status regulate csgD expression on a post-transcriptional level by unknown mechanisms. Furthermore, phosphorylation of CsgD is a potential inactivation and degradation signal in biofilm dissolution. Including complex incoherent feed-forward loops, regulation of biofilm formation versus motility and virulence is of recognized complexity.

  9. Deletion of Salmonella enterica serovar typhimurium sipC gene

    Institute of Scientific and Technical Information of China (English)

    Maryam Safarpour Dehkordi; Abbas Doosti; Asghar Arshi

    2015-01-01

    Objective: To construct a novel plasmid as Salmonella enterica serovar typhimurium (S. typhimurium) sipC gene knockouts candidate. Methods: In this research, 50 upstream and 30 downstream regions of S. typhimurium sipC gene and kanamycin gene were PCR amplified. Each of these DNA fragment was cloned into pGEM T-easy vector. The construct was confirmed by PCR and restriction digest. Results: PCR amplified 320, 206 and 835 bp DNA fragments were subcloned into pET-32 vector resulting with a plasmid called pET-32-sipC up-kan-sip C down. Conclusions: The new plasmid (pET-32-sipC up-kan-sip C down) is useful for genetic engineering and for future manipulation of S. typhimurium sipC gene.

  10. Deletion of Salmonella enterica serovar typhimurium sipC gene

    Institute of Scientific and Technical Information of China (English)

    Maryam; Safarpour; Dehkordi; Abbas; Doosti; Asghar; Arshi

    2015-01-01

    Objective:To construct a novel plasmid as Salmonella enterica serovar typhimurium(S.typhimurium)sip C gene knockouts candidate.Methods:In this research,50upstream and 30downstream regions of S.typhimurium sip C gene and kanamycin gene were PCR amplified.Each of these DNA fragment was cloned into p GEM T-easy vector.The construct was confirmed by PCR and restriction digest.Results:PCR amplified 320,206 and 835 bp DNA fragments were subcloned into p ET-32 vector resulting with a plasmid called p ET-32-sip C up-kan-sip C down.Conclusions:The new plasmid(p ET-32-sip C up-kan-sip C down)is useful for genetic engineering and for future manipulation of S.typhimurium sip C gene.

  11. Genomic variation in Salmonella enterica core genes for epidemiological typing

    DEFF Research Database (Denmark)

    Leekitcharoenphon, Pimlapas; Lukjancenko, Oksana; Rundsten, Carsten Friis;

    2012-01-01

    genomes and evaluate their value as typing targets, comparing whole genome typing and traditional methods such as 16S and MLST. A consensus tree based on variation of core genes gives much better resolution than 16S and MLST; the pan-genome family tree is similar to the consensus tree, but with higher......Background: Technological advances in high throughput genome sequencing are making whole genome sequencing (WGS) available as a routine tool for bacterial typing. Standardized procedures for identification of relevant genes and of variation are needed to enable comparison between studies and over...... time. The core genes-the genes that are conserved in all (or most) members of a genus or species-are potentially good candidates for investigating genomic variation in phylogeny and epidemiology. Results: We identify a set of 2,882 core genes clusters based on 73 publicly available Salmonella enterica...

  12. Biofilm Formation and Morphotypes of Salmonella enterica subsp.arizonae Differs from Those of Other Salmonella enterica Subspecies in Isolates from Poultry Houses.

    Science.gov (United States)

    Lamas, A; Fernandez-No, I C; Miranda, J M; Vázquez, B; Cepeda, A; Franco, C M

    2016-07-01

    Salmonella serovars are responsible for foodborne diseases around the world. The ability to form biofilms allows microorganisms to survive in the environment. In this study, 73 Salmonella strains, belonging to four different subspecies, were isolated from poultry houses and foodstuffs and tested. Biofilm formation was measured at four different temperatures and two nutrient concentrations. Morphotypes and cellulose production were evaluated at three different temperatures. The presence of several genes related to biofilm production was also examined. All strains and subspecies of Salmonella had the ability to form biofilms, and 46.57% of strains produced biofilms under all conditions tested. Biofilm formation was strain dependent and varied according to the conditions. This is the first study to analyze biofilm formation in a wide number of Salmonella enterica subsp. arizonae strains, and no direct relationship between the high prevalence of Salmonella enterica subsp. arizonae strains and their ability to form biofilm was established. Morphotypes and cellulose production varied as the temperature changed, with 20°C being the optimum temperature for expression of the red, dry, and rough morphotype and cellulose. Salmonella enterica subsp. arizonae, whose morphotype is poorly studied, only showed a smooth and white morphotype and lacked the csgD and gcpA genes that are implicated in biofilm production. Thus, Salmonella biofilm formation under different environmental conditions is a public health problem because it can survive and advance through the food chain to reach the consumer.

  13. Regulation of Salmonella enterica pathogenicity island 1 by DNA adenine methylation.

    Science.gov (United States)

    López-Garrido, Javier; Casadesús, Josep

    2010-03-01

    DNA adenine methylase (Dam(-)) mutants of Salmonella enterica are attenuated in the mouse model and present multiple virulence-related defects. Impaired interaction of Salmonella Dam(-) mutants with the intestinal epithelium has been tentatively correlated with reduced secretion of pathogenicity island 1 (SPI-1) effectors. In this study, we show that S. enterica Dam(-) mutants contain lowered levels of the SPI-1 transcriptional regulators HilA, HilC, HilD, and InvF. Epistasis analysis indicates that Dam-dependent regulation of SPI-1 requires HilD, while HilA, HilC, and InvF are dispensable. A transcriptional hilDlac fusion is expressed at similar levels in Dam(+) and Dam(-) hosts. However, lower levels of hilD mRNA are found in a Dam(-) background, thus providing unsuspected evidence that Dam methylation might exert post-transcriptional regulation of hilD expression. This hypothesis is supported by the following lines of evidence: (i) lowered levels of hilD mRNA are found in Salmonella Dam(-) mutants when hilD is transcribed from a heterologous promoter; (ii) increased hilD mRNA turnover is observed in Dam(-) mutants; (iii) lack of the Hfq RNA chaperone enhances hilD mRNA instability in Dam(-) mutants; and (iv) lack of the RNA degradosome components polynucleotide phosphorylase and ribonuclease E suppresses hilD mRNA instability in a Dam(-) background. Our report of Dam-dependent control of hilD mRNA stability suggests that DNA adenine methylation plays hitherto unknown roles in post-transcriptional control of gene expression.

  14. The anti-infective activity of punicalagin against Salmonella enterica subsp. enterica serovar typhimurium in mice.

    Science.gov (United States)

    Li, Guanghui; Feng, Yuqing; Xu, Yunfeng; Wu, Qian; Han, Qi'an; Liang, Xiujun; Yang, Baowei; Wang, Xin; Xia, Xiaodong

    2015-07-01

    Punicalagin, a major bioactive component of pomegranate peel, has been proven to have antioxidant, antiviral, anti-apoptosis, and hepatoprotective properties. The aim of this study was to investigate the anti-infective activity of punicalagin in a mouse model. C57BL/6 mice were initially challenged with Salmonella enterica subsp. enterica serovar typhimurium (S. typhimurium) and then treated with punicalagin. Food and water consumption and body weight were recorded daily. On day 8 post infection, the mice were sacrificed to examine pathogen counts in tissues, hematological parameters, cytokine levels, and histological changes. Compared to mice only infected with S. typhimurium, punicalagin-treated mice had more food consumption and less weight loss. A higher survival rate and lower counts of viable S. typhimurium in feces, liver, spleen, and kidney were found in the punicalagin-treated mice. The enzyme linked immunosorbent assay showed that the levels of IL-6, IL-10, and IFN-γ in serum and the spleen and TNF-α in serum, the spleen and the liver were reduced by punicalagin. Moreover, more neutrophils and higher neutrophil-to-mononuclear cell ratios in the punicalagin-treated mice were observed. Histological examination showed that punicalagin protected cells in the liver and spleen from hemorrhagic necrosis. It is concluded that punicalagin has a beneficial effect against S. typhimurium infection in mice. The anti-infective properties, together with other nutritionally beneficial effects, make punicalagin a promising supplement in human food or animal feeds to prevent disease associated with S. typhimurium.

  15. 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.

  16. Intracellular demography and the dynamics of Salmonella enterica infections.

    Directory of Open Access Journals (Sweden)

    Sam P Brown

    2006-10-01

    Full Text Available An understanding of within-host dynamics of pathogen interactions with eukaryotic cells can shape the development of effective preventive measures and drug regimes. Such investigations have been hampered by the difficulty of identifying and observing directly, within live tissues, the multiple key variables that underlay infection processes. Fluorescence microscopy data on intracellular distributions of Salmonella enterica serovar Typhimurium (S. Typhimurium show that, while the number of infected cells increases with time, the distribution of bacteria between cells is stationary (though highly skewed. Here, we report a simple model framework for the intensity of intracellular infection that links the quasi-stationary distribution of bacteria to bacterial and cellular demography. This enables us to reject the hypothesis that the skewed distribution is generated by intrinsic cellular heterogeneities, and to derive specific predictions on the within-cell dynamics of Salmonella division and host-cell lysis. For within-cell pathogens in general, we show that within-cell dynamics have implications across pathogen dynamics, evolution, and control, and we develop novel generic guidelines for the design of antibacterial combination therapies and the management of antibiotic resistance.

  17. Cellular requirements for systemic control of Salmonella enterica serovar Typhimurium infections in mice.

    Science.gov (United States)

    Kupz, Andreas; Bedoui, Sammy; Strugnell, Richard A

    2014-12-01

    The rational design of vaccines requires an understanding of the contributions of individual immune cell subsets to immunity. With this understanding, targeted vaccine delivery approaches and adjuvants can be developed to maximize vaccine efficiency and to minimize side effects (S. H. E. Kaufmann et al., Immunity 33:555-577, 2010; T. Ben-Yedidia and R. Arnon, Hum. Vaccines 1:95-101, 2005). We have addressed the contributions of different immune cell subsets and their ability to contribute to the control and clearance of the facultative intracellular pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium) in a murine model. Using a systematic and reproducible model of experimental attenuated S. Typhimurium infection, we show that distinct lymphocyte deficiencies lead to one of four different infection outcomes: clearance, chronic infection, early death, or late death. Our study demonstrates a high level of functional redundancy in the ability of different lymphocyte subsets to provide interferon gamma (IFN-γ), a critical cytokine in Salmonella immunity. Whereas early control of the infection was entirely dependent on IFN-γ but not on any particular lymphocyte subset, clearance of the infection critically required CD4(+) T cells but appeared to be independent of IFN-γ. These data reinforce the idea of a bimodal immune response against Salmonella: an early T cell-independent but IFN-γ-dependent phase and a late T cell-dependent phase that may be IFN-γ independent. PMID:25225248

  18. International Spread of an Epidemic Population of Salmonella enterica Serotype Kentucky ST198 Resistant to Ciprofloxacin

    DEFF Research Database (Denmark)

    Le Hello, Simon; Hendriksen, Rene S.; Doublet, Benoit;

    2011-01-01

    National Salmonella surveillance systems from France, England and Wales, Denmark, and the United States identified the recent emergence of multidrug-resistant isolates of Salmonella enterica serotype Kentucky displaying high-level resistance to ciprofloxacin. A total of 489 human cases were ident...

  19. Salmonella enterica Serovar Virchow Bacteremia Presenting as Typhoid-Like Illness in an Immunocompetent Patient ▿

    OpenAIRE

    Eckerle, Isabella; Zimmermann, Stefan; Kapaun, Annette; Junghanss, Thomas

    2010-01-01

    We report a typhoid-like illness with fever and altered consciousness in a 22-year-old man with growth of Salmonella enterica serovar Virchow in blood and stool culture. Bacteremia and invasive disease due to non-typhoid salmonellae (NTS) are known in severely immunocompromised patients, but so far have not been described in immunocompetent adults.

  20. Draft Genome Sequences of 11 Salmonella enterica Strains with Variable Levels of Barotolerance

    Science.gov (United States)

    Ronholm, Jennifer; Petronella, Nicholas

    2016-01-01

    The diversity of the genus Salmonella is reflected in the physiological adaptations used by its members in response to stressors such as high pressure. Here we report the draft whole genome sequences of 11 Salmonella enterica strains, five sensitive strains and six demonstrating high levels of pressure resistance. PMID:27660773

  1. New Flagellin Gene for Salmonella enterica serovar Typhi from the East Indonesian Archipelago

    NARCIS (Netherlands)

    M. Hatta; A.R. Sultan; R. Pastoor; H.L. Smits

    2011-01-01

    Phase variation is a property unique of some Salmonella enterica serovar Typhi strains from Indonesia. Salmonella Typhi isolates from Indonesia have been described that in addition to the phase 1 Hd flagellin gene contain a second flagellin gene named z66. S. Typhi isolates from Indonesia with a mut

  2. Draft Genome Sequences of 11 Salmonella enterica Strains with Variable Levels of Barotolerance.

    Science.gov (United States)

    Ronholm, Jennifer; Petronella, Nicholas; Tamber, Sandeep

    2016-01-01

    The diversity of the genus Salmonella is reflected in the physiological adaptations used by its members in response to stressors such as high pressure. Here we report the draft whole genome sequences of 11 Salmonella enterica strains, five sensitive strains and six demonstrating high levels of pressure resistance. PMID:27660773

  3. Intragastric immunization with recombinant Lactobacillus casei expressing flagellar antigen confers antibody-independent protective immunity against Salmonella enterica serovar Enteritidis

    NARCIS (Netherlands)

    Kajikawa, A.; Satoh, E.; Leer, R.J.; Yamamoto, S.; Igimi, S.

    2007-01-01

    A recombinant Lactobacillus casei expressing a flagellar antigen from Salmonella enterica serovar Enteritidis was constructed and evaluated as a mucosal vaccine. Intragastric immunization of the recombinant strain conferred protective immunity against Salmonella infection in mice. This immunization

  4. 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.

  5. Oral infection of mice with Salmonella enterica serovar Typhimurium causes meningitis and infection of the brain

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    Provias John

    2007-06-01

    Full Text Available Abstract Background Salmonella meningitis is a rare and serious infection of the central nervous system following acute Salmonella enterica sepsis. For this pathogen, no appropriate model has been reported in which to examine infection kinetics and natural dissemination to the brain. Methods Five mouse lines including C57BL/6, Balb/c, 129S6-Slc11a1tm1Mcg, 129S1/SvImJ, B6.129-Inpp5dtm1Rkh were used in the murine typhoid model to examine the dissemination of systemic Salmonella enterica serovar Typhimurium following oral infection. Results We report data on spontaneous meningitis and brain infection following oral infection of mice with Salmonella enterica serovar Typhimurium. Conclusion This model may provide a system in which dissemination of bacteria through the central nervous system and the influence of host and bacterial genetics can be queried.

  6. Salmonella enterica Serotype Choleraesuis Infection of the Knee and Femur in a Nonbacteremic Diabetic Patient

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    Alexander M. Sy

    2013-01-01

    Full Text Available Osteoarticular infections caused by Salmonella are rare. The rates of osteomyelitis and septic arthritis due to Salmonella are estimated to be less than 1% and 0.1%-0.2%, respectively (Kato et al., 2012. Salmonella enterica serotype Choleraesuis is a nontyphoidal Salmonella, highly pathogenic in humans, usually causing septicemic disease with little or no intestinal involvement. Serotype Choleraesuis accounts for a small percentage of published studies of Salmonella infections in the United States. It is not commonly reported in joint fluid and bones in contrast to serotype Enteritidis and Typhi, where a considerable number of cases have been published. Chen et al. in Taiwan found that 21% of bacteremic patients with this infection subsequently develop focal infections such as septic arthritis, pneumonia, peritonitis, and cutaneous abscess (Chen et al., 1999, Chiu et al., 2004. In contrast, our patient presented with localized osteoarticular infection with Salmonella enterica serotype Cholerasuis, but without evidence of bacteremia.

  7. The Salmonella pathogenicity island 2-encoded type III secretion system is essential for the survival of Salmonella enterica serovar Typhimurium in free-living amoebae.

    Science.gov (United States)

    Bleasdale, Benjamin; Lott, Penelope J; Jagannathan, Aparna; Stevens, Mark P; Birtles, Richard J; Wigley, Paul

    2009-03-01

    Free-living amoebae represent a potential reservoir and predator of Salmonella enterica. Through the use of type III secretion system (T3SS) mutants and analysis of transcription of selected T3SS genes, we demonstrated that the Salmonella pathogenicity island 2 is highly induced during S. enterica serovar Typhimurium infection of Acanthamoeba polyphaga and is essential for survival within amoebae.

  8. Tandem attenuators control expression of the Salmonella mgtCBR virulence operon

    OpenAIRE

    Lee, Eun-Jin; Groisman, Eduardo A.

    2012-01-01

    The mgtCBR operon from Salmonella enterica serovar Typhimurium specifies the virulence protein MgtC, the Mg2+ transporter MgtB and the regulatory peptide MgtR. The mgtCBR transcript includes a long leader region harbouring two short open reading frames (ORFs). Translation of these ORFs is anticipated to impact the formation of particular stem-loop structures and control transcription of the coding region by an attenuation-like mechanism. We previously reported that ORF mgtM enables Salmonella...

  9. 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.

  10. Effects of integrated treatment of nonthermal UV-C light and different antimicrobial wash on Salmonella enterica on plum tomatoes

    Science.gov (United States)

    Produce contamination by foodborne pathogens remains a serious threat. This study investigated synergistic effects of ultraviolet-C and various active sanitizers’ washes against Salmonella enterica on plum tomatoes. A bacterial cocktail containing three serotypes of Salmonella enterica (S. Newport H...

  11. Application of molecular methods for identification of strains classified as Salmonella enterica serovar 6, 7/-/- by conventional serotyping

    DEFF Research Database (Denmark)

    Chadfield, M. S.; Christensen, J. P.; Madsen, Mogens;

    2002-01-01

    An increased prevalence of Salmonella enterica serovar Tennessee (6, 7: z(29):-) was observed in broiler flocks in Denmark in 1994 and a parallel increase in the prevalence of Salmonella enterica serovar 6, 7:-:- was demonstrated, albeit at a lower level. Plasmid profiling and ribotyping revealed...

  12. Survival of Salmonella enterica in poultry feed is strain dependent.

    Science.gov (United States)

    Andino, Ana; Pendleton, Sean; Zhang, Nan; Chen, Wei; Critzer, Faith; Hanning, Irene

    2014-02-01

    Feed components have low water activity, making bacterial survival difficult. The mechanisms of Salmonella survival in feed and subsequent colonization of poultry are unknown. The purpose of this research was to compare the ability of Salmonella serovars and strains to survive in broiler feed and to evaluate molecular mechanisms associated with survival and colonization by measuring the expression of genes associated with colonization (hilA, invA) and survival via fatty acid synthesis (cfa, fabA, fabB, fabD). Feed was inoculated with 1 of 15 strains of Salmonella enterica consisting of 11 serovars (Typhimurium, Enteriditis, Kentucky, Seftenburg, Heidelberg, Mbandanka, Newport, Bairely, Javiana, Montevideo, and Infantis). To inoculate feed, cultures were suspended in PBS and survival was evaluated by plating samples onto XLT4 agar plates at specific time points (0 h, 4 h, 8 h, 24 h, 4 d, and 7 d). To evaluate gene expression, RNA was extracted from the samples at the specific time points (0, 4, 8, and 24 h) and gene expression measured with real-time PCR. The largest reduction in Salmonella occurred at the first and third sampling time points (4 h and 4 d) with the average reductions being 1.9 and 1.6 log cfu per g, respectively. For the remaining time points (8 h, 24 h, and 7 d), the average reduction was less than 1 log cfu per g (0.6, 0.4, and 0.6, respectively). Most strains upregulated cfa (cyclopropane fatty acid synthesis) within 8 h, which would modify the fluidity of the cell wall to aid in survival. There was a weak negative correlation between survival and virulence gene expression indicating downregulation to focus energy on other gene expression efforts such as survival-related genes. These data indicate the ability of strains to survive over time in poultry feed was strain dependent and that upregulation of cyclopropane fatty acid synthesis and downregulation of virulence genes were associated with a response to desiccation stress.

  13. Genomic variation in Salmonella enterica core genes for epidemiological typing

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    Leekitcharoenphon Pimlapas

    2012-03-01

    Full Text Available Abstract Background Technological advances in high throughput genome sequencing are making whole genome sequencing (WGS available as a routine tool for bacterial typing. Standardized procedures for identification of relevant genes and of variation are needed to enable comparison between studies and over time. The core genes--the genes that are conserved in all (or most members of a genus or species--are potentially good candidates for investigating genomic variation in phylogeny and epidemiology. Results We identify a set of 2,882 core genes clusters based on 73 publicly available Salmonella enterica genomes and evaluate their value as typing targets, comparing whole genome typing and traditional methods such as 16S and MLST. A consensus tree based on variation of core genes gives much better resolution than 16S and MLST; the pan-genome family tree is similar to the consensus tree, but with higher confidence. The core genes can be divided into two categories: a few highly variable genes and a larger set of conserved core genes, with low variance. For the most variable core genes, the variance in amino acid sequences is higher than for the corresponding nucleotide sequences, suggesting that there is a positive selection towards mutations leading to amino acid changes. Conclusions Genomic variation within the core genome is useful for investigating molecular evolution and providing candidate genes for bacterial genome typing. Identification of genes with different degrees of variation is important especially in trend analysis.

  14. Mapping the Regulatory Network for Salmonella enterica Serovar Typhimurium Invasion

    Science.gov (United States)

    Smith, Carol; Stringer, Anne M.; Mao, Chunhong; Palumbo, Michael J.

    2016-01-01

    ABSTRACT Salmonella enterica pathogenicity island 1 (SPI-1) encodes proteins required for invasion of gut epithelial cells. The timing of invasion is tightly controlled by a complex regulatory network. The transcription factor (TF) HilD is the master regulator of this process and senses environmental signals associated with invasion. HilD activates transcription of genes within and outside SPI-1, including six other TFs. Thus, the transcriptional program associated with host cell invasion is controlled by at least 7 TFs. However, very few of the regulatory targets are known for these TFs, and the extent of the regulatory network is unclear. In this study, we used complementary genomic approaches to map the direct regulatory targets of all 7 TFs. Our data reveal a highly complex and interconnected network that includes many previously undescribed regulatory targets. Moreover, the network extends well beyond the 7 TFs, due to the inclusion of many additional TFs and noncoding RNAs. By comparing gene expression profiles of regulatory targets for the 7 TFs, we identified many uncharacterized genes that are likely to play direct roles in invasion. We also uncovered cross talk between SPI-1 regulation and other regulatory pathways, which, in turn, identified gene clusters that likely share related functions. Our data are freely available through an intuitive online browser and represent a valuable resource for the bacterial research community. PMID:27601571

  15. Increased water activity reduces the thermal resistance of Salmonella enterica in peanut butter.

    Science.gov (United States)

    He, Yingshu; Li, Ye; Salazar, Joelle K; Yang, Jingyun; Tortorello, Mary Lou; Zhang, Wei

    2013-08-01

    Increased water activity in peanut butter significantly (P < 0.05) reduced the heat resistance of desiccation-stressed Salmonella enterica serotypes treated at 90 °C. The difference in thermal resistance was less notable when strains were treated at 126 °C. Using scanning electron microscopy, we observed minor morphological changes of S. enterica cells resulting from desiccation and rehydration processes in peanut oil. PMID:23728806

  16. Isolation and characterization of Salmonella enterica in day-old ducklings in Egypt.

    Science.gov (United States)

    Osman, Kamelia M; Marouf, Sherif H; Zolnikov, Tara R; AlAtfeehy, Nayerah

    2014-01-01

    Importing day-old ducklings (DOD) unknowingly infected with non-typhoid Salmonella (NTS) may be associated with disease risk. Domestic and international trade may enhance this risk. Salmonella enterica serovars, their virulence genes combinations and antibiotic resistance, garner attention for their potentiality to contribute to the adverse health effects on populations throughout the world. The aim of this study was to estimate the risk of imported versus domestic DOD as potential carriers of NTS. The results confirm the prevalence of salmonellosis in imported ducklings was 18·5% (25/135), whereas only 12% (9/75) of cases were determined in the domestic ducklings. Fourteen serovars (Salmonella enteritidis, Salmonella kisii, Salmonella typhimurium, Salmonella gaillac, Salmonella uno, Salmonella eingedi, Salmonella shubra, Salmonella bardo, Salmonella inganda, Salmonella kentucky, Salmonella stanley, Salmonella virchow, Salmonella haifa, and Salmonella anatum) were isolated from the imported ducklings, whereas only S. enteritidis, S. typhimurium, S. virchow, and S. shubra were isolated from the domestic ducklings. The isolated Salmonella serovars were 100% susceptible to only colistin sulphate and 100% resistant to lincomycin. The 14 Salmonella serovars were screened for 11 virulence genes (invA, avrA, ssaQ, mgtC, siiD, sopB, gipA, sodC1, sopE1, spvC, and bcfC) by PCR. The invA, sopB, and bcfC genes were detected in 100% of the Salmonella serovars; alternatively, the gipA gene was absent in all of the isolated Salmonella serovars. The 11 virulent genes were not detected in either of S. stanley or S. haifa serovars. The results confirm an association between antibiotic resistance and virulence of Salmonella in the DOD. This study confirms the need for a country adherence to strict public health and food safety regimes.

  17. Molecular typing of Salmonella enterica subspecies enterica serovar Typhimurium isolated in Abruzzo region (Italy from 2008 to 2010

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    Alessandra Alessiani

    2014-03-01

    Full Text Available In this study, 47 antibiotic-resistant strains of Salmonella enterica subspecies enterica serovar Typhimurium (ST were characterised, including 15 monophasic variants 1, 4, [5], 12:i:-, (STm isolated from different matrices. They were all selected from 389 Salmonella enterica subspecies enterica strains isolated during 2008-2010 in Abruzzo region (Italy. Thirty-seven strains showed to be resistant to more than 1 antibiotic. Among 47 isolates, phage type U311 and DT104 were identified. The ASSuT resistance pattern was predominant in mST strains and ACSSuT in ST DT104 and U302. A multiplex Polimerase Chain Reaction (PCR method was used to investigate 4 genes: fluorfenicol (floSt, virulence (spvC, invasine (invA and integrase (int. All ST the strain were positive for invA gene and 28,32% of strains were positive for spvC gene. PFGE analysis revealed a large number of small clonal populations, however not ascrivable to outbreaks.

  18. Molecular typing of Salmonella enterica subspecies enterica serovar Typhimurium isolated in Abruzzo region (Italy) from 2008 to 2010.

    Science.gov (United States)

    Alessiani, Alessandra; Sacchini, Lorena; Pontieri, Eugenio; Gavini, Jacopo; Di Giannatale, Elisabetta

    2014-01-01

    In this study, 47 antibiotic-resistant strains of Salmonella enterica subspecies enterica serovar Typhimurium (ST) were characterised, including 15 monophasic variants 1, 4, [5], 12:i:-, (STm) isolated from different matrices. They were all selected from 389 Salmonella enterica subspecies enterica strains isolated during 2008-2010 in Abruzzo region (Italy). Thirty-seven strains showed to be resistant to more than 1 antibiotic. Among 47 isolates, phage type U311 and DT104 were identified. The ASSuT resistance pattern was predominant in mST strains and ACSSuT in ST DT104 and U302. A multiplex Polimerase Chain Reaction (PCR) method was used to investigate 4 genes: fluorfenicol (floSt), virulence (spvC), invasine (invA) and integrase (int). All ST the strain were positive for invA gene and 28,32% of strains were positive for spvC gene. PFGE analysis revealed a large number of small clonal populations, however not ascrivable to outbreaks.

  19. Complete Genome and Plasmid Sequences of Three Canadian Strains of Salmonella enterica subsp. enterica Serovar Enteritidis Belonging to Phage Types 8, 13, and 13a

    Science.gov (United States)

    Rehman, Muhammad Attiq; Labbé, Geneviève; Ziebell, Kim; Nash, John H. E.

    2015-01-01

    Salmonella enterica subsp. enterica serovar Enteritidis is a prominent cause of human salmonellosis frequently linked to poultry products. In Canada, S. Enteritidis phage types 8, 13, and 13a predominate among both clinical and poultry isolates. Here, we report the complete genome and plasmid sequences of poultry isolates of these three phage types. PMID:26404595

  20. Salmonella Enterica Serotype Enteritidis Vertebral Osteomyelitis and Epidural Abscess Complicated with Meningitis.

    Science.gov (United States)

    Oki, Masayuki; Ueda, Akihiro; Tsuda, Ayumi; Yanagi, Hidetaka; Ozawa, Hideki; Takagi, Atsushi

    2016-01-01

    Infection with non-typhoidal Salmonella often results in a self-limited acute gastroenteritis. Extra-intestinal Salmonella infection is relatively rare and occurs predominantly in infants and adults with significant underlying conditions. We describe a 54-year-old Japanese man with a history of heavy alcohol consumption and daily contact with a dog, who developed bacteremia complicated by vertebral osteomyelitis, spinal epidural abscess, and meningitis, due to Salmonella enterica serotype Enteritidis. This case suggests that Salmonella should be considered as an etiologic pathogen in adult patients with perivertebral infection or meningitis. PMID:27628612

  1. Genomic characterisation of invasive non-typhoidal Salmonella enterica Subspecies enterica Serovar Bovismorbificans isolates from Malawi.

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    Christina Bronowski

    2013-11-01

    Full Text Available BACKGROUND: 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. METHODOLOGY: 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. PRINCIPAL FINDINGS: 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. CONCLUSIONS: 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

  2. Comparative proteomic analysis of the PhoP regulon in Salmonella enterica serovar Typhi versus Typhimurium.

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    Richelle C Charles

    Full Text Available BACKGROUND: S. Typhi, a human-restricted Salmonella enterica serovar, causes a systemic intracellular infection in humans (typhoid fever. In comparison, S. Typhimurium causes gastroenteritis in humans, but causes a systemic typhoidal illness in mice. The PhoP regulon is a well studied two component (PhoP/Q coordinately regulated network of genes whose expression is required for intracellular survival of S. enterica. METHODOLOGY/PRINCIPAL FINDINGS: Using high performance liquid chromatography mass spectrometry (HPLC-MS/MS, we examined the protein expression profiles of three sequenced S. enterica strains: S. Typhimurium LT2, S. Typhi CT18, and S. Typhi Ty2 in PhoP-inducing and non-inducing conditions in vitro and compared these results to profiles of phoP(-/Q(- mutants derived from S. Typhimurium LT2 and S. Typhi Ty2. Our analysis identified 53 proteins in S. Typhimurium LT2 and 56 proteins in S. Typhi that were regulated in a PhoP-dependent manner. As expected, many proteins identified in S. Typhi demonstrated concordant differential expression with a homologous protein in S. Typhimurium. However, three proteins (HlyE, STY1499, and CdtB had no homolog in S. Typhimurium. HlyE is a pore-forming toxin. STY1499 encodes a stably expressed protein of unknown function transcribed in the same operon as HlyE. CdtB is a cytolethal distending toxin associated with DNA damage, cell cycle arrest, and cellular distension. Gene expression studies confirmed up-regulation of mRNA of HlyE, STY1499, and CdtB in S. Typhi in PhoP-inducing conditions. CONCLUSIONS/SIGNIFICANCE: This study is the first protein expression study of the PhoP virulence associated regulon using strains of Salmonella mutant in PhoP, has identified three Typhi-unique proteins (CdtB, HlyE and STY1499 that are not present in the genome of the wide host-range Typhimurium, and includes the first protein expression profiling of a live attenuated bacterial vaccine studied in humans (Ty800.

  3. Role of soil, crop debris, and a plant pathogen in Salmonella enterica contamination of tomato plants.

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    Jeri D Barak

    Full Text Available BACKGROUND: In the U.S., tomatoes have become the most implicated vehicle for produce-associated Salmonellosis with 12 outbreaks since 1998. Although unconfirmed, trace backs suggest pre-harvest contamination with Salmonella enterica. Routes of tomato crop contamination by S. enterica in the absence of direct artificial inoculation have not been investigated. METHODOLOGY/PRINCIPAL FINDINGS: This work examined the role of contaminated soil, the potential for crop debris to act as inoculum from one crop to the next, and any interaction between the seedbourne plant pathogen Xanthomonas campestris pv. vesicatoria and S. enterica on tomato plants. Our results show S. enterica can survive for up to six weeks in fallow soil with the ability to contaminate tomato plants. We found S. enterica can contaminate a subsequent crop via crop debris; however a fallow period between crop incorporation and subsequent seeding can affect contamination patterns. Throughout these studies, populations of S. enterica declined over time and there was no bacterial growth in either the phyllosphere or rhizoplane. The presence of X. campestris pv. vesicatoria on co-colonized tomato plants had no effect on the incidence of S. enterica tomato phyllosphere contamination. However, growth of S. enterica in the tomato phyllosphere occurred on co-colonized plants in the absence of plant disease. CONCLUSIONS/SIGNIFICANCE: S. enterica contaminated soil can lead to contamination of the tomato phyllosphere. A six week lag period between soil contamination and tomato seeding did not deter subsequent crop contamination. In the absence of plant disease, presence of the bacterial plant pathogen, X. campestris pv. vesicatoria was beneficial to S. enterica allowing multiplication of the human pathogen population. Any event leading to soil contamination with S. enterica could pose a public health risk with subsequent tomato production, especially in areas prone to bacterial spot disease.

  4. Variable carbon catabolism among Salmonella enterica serovar Typhi isolates.

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    Lay Ching Chai

    Full Text Available BACKGROUND: Salmonella enterica serovar Typhi (S. Typhi is strictly a human intracellular pathogen. It causes acute systemic (typhoid fever and chronic infections that result in long-term asymptomatic human carriage. S. Typhi displays diverse disease manifestations in human infection and exhibits high clonality. The principal factors underlying the unique lifestyle of S. Typhi in its human host during acute and chronic infections remain largely unknown and are therefore the main objective of this study. METHODOLOGY/PRINCIPAL FINDINGS: To obtain insight into the intracellular lifestyle of S. Typhi, a high-throughput phenotypic microarray was employed to characterise the catabolic capacity of 190 carbon sources in S. Typhi strains. The success of this study lies in the carefully selected library of S. Typhi strains, including strains from two geographically distinct areas of typhoid endemicity, an asymptomatic human carrier, clinical stools and blood samples and sewage-contaminated rivers. An extremely low carbon catabolic capacity (27% of 190 carbon substrates was observed among the strains. The carbon catabolic profiles appeared to suggest that S. Typhi strains survived well on carbon subtrates that are found abundantly in the human body but not in others. The strains could not utilise plant-associated carbon substrates. In addition, α-glycerolphosphate, glycerol, L-serine, pyruvate and lactate served as better carbon sources to monosaccharides in the S. Typhi strains tested. CONCLUSION: The carbon catabolic profiles suggest that S. Typhi could survive and persist well in the nutrient depleted metabolic niches in the human host but not in the environment outside of the host. These findings serve as caveats for future studies to understand how carbon catabolism relates to the pathogenesis and transmission of this pathogen.

  5. Eggshell penetration of hen's eggs by Salmonella enterica serovar Enteritidis upon various storage conditions

    OpenAIRE

    Messens, Winy; Grijspeerdt, Koen; Herman, Lieve

    2006-01-01

    Abstract 1. The survival and penetration of Salmonella enterica serovar Enteritidis (SE) inoculated on the eggshell was examined upon storage for up to 20 d at real-life conditions (15?C to 25?C and 45% to 75% relative humidity (RH)). 2. Penetration was assessed by emptying the egg contents and filling the eggs with a selective medium that allowed visualising Salmonella growth on the inside of the shell and membrane complex. 3. The study of survival on the eggshells was base...

  6. Dissemination of antimicrobial-resistant clones of Salmonella enterica among domestic animals, wild animals, and humans.

    Science.gov (United States)

    Palomo, Gonzalo; Campos, Maria Jorge; Ugarte, María; Porrero, María Concepción; Alonso, Juan Manuel; Borge, Carmen; Vadillo, Santiago; Domínguez, Lucas; Quesada, Alberto; Píriz, Segundo

    2013-02-01

    Non-typhoidal salmonellosis is an important zoonotic disease caused by Salmonella enterica. This work focuses on the identification of Salmonella enterica clonal strains which, presenting a wide distribution potential, express resistance determinants that compromise effectiveness of the antimicrobial therapy. The screening was performed on 506 Salmonella enterica isolates from animals and humans, which were characterized by serovar and phage typing, genome macrorestriction and pulsed-field gel electrophoresis, and detection of phenotypic and genotypic traits for antimicrobial resistance. A Salmonella Enteritidis strain with strong quinolone resistance is spread on three host environments carrying one of the four variants found for the GyrA protein: (1) Asp87Tyr, the major polymorphism found in 39 Salmonella isolates from human origin and six from poultry; (2) Ser83Phe, with four isolates from human origin and one from white stork (Ciconia ciconia); and (3) Asp87Asn or (4) Asp87Gly, with two isolates each from human origins. Several Salmonella Typhimurium strains that presented int1 elements and the classically associated pentaresistance (ACSSuT) phenotype were found distributed between two host environments: domestic animals and humans, domestics and wild animals, or wild fauna plus humans. This study points out the importance of monitoring gut microbiota and its antimicrobial resistance from wildlife, in parallel to livestock animals and humans, especially for animal species that are in close contact with people. PMID:23360170

  7. Dissemination of antimicrobial-resistant clones of Salmonella enterica among domestic animals, wild animals, and humans.

    Science.gov (United States)

    Palomo, Gonzalo; Campos, Maria Jorge; Ugarte, María; Porrero, María Concepción; Alonso, Juan Manuel; Borge, Carmen; Vadillo, Santiago; Domínguez, Lucas; Quesada, Alberto; Píriz, Segundo

    2013-02-01

    Non-typhoidal salmonellosis is an important zoonotic disease caused by Salmonella enterica. This work focuses on the identification of Salmonella enterica clonal strains which, presenting a wide distribution potential, express resistance determinants that compromise effectiveness of the antimicrobial therapy. The screening was performed on 506 Salmonella enterica isolates from animals and humans, which were characterized by serovar and phage typing, genome macrorestriction and pulsed-field gel electrophoresis, and detection of phenotypic and genotypic traits for antimicrobial resistance. A Salmonella Enteritidis strain with strong quinolone resistance is spread on three host environments carrying one of the four variants found for the GyrA protein: (1) Asp87Tyr, the major polymorphism found in 39 Salmonella isolates from human origin and six from poultry; (2) Ser83Phe, with four isolates from human origin and one from white stork (Ciconia ciconia); and (3) Asp87Asn or (4) Asp87Gly, with two isolates each from human origins. Several Salmonella Typhimurium strains that presented int1 elements and the classically associated pentaresistance (ACSSuT) phenotype were found distributed between two host environments: domestic animals and humans, domestics and wild animals, or wild fauna plus humans. This study points out the importance of monitoring gut microbiota and its antimicrobial resistance from wildlife, in parallel to livestock animals and humans, especially for animal species that are in close contact with people.

  8. 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...

  9. Impact of Strain Variation on the Ability of Biosensor Technology to Detect Salmonella enterica

    Science.gov (United States)

    Introduction: It is important to develop methods that can quickly and accurately detect the presence of bacteria in the food supply that cause disease. Salmonella enterica is a bacteria that is often associated with contamination of food. Strains vary in their ability to cause illness and to spread...

  10. Outbreak-associated Salmonella enterica Serotypes and Food Commodities, United States, 1998- 2008

    Centers for Disease Control (CDC) Podcasts

    2013-09-09

    Dr. Mike Miller reads an abridged version of the Emerging Infectious Diseases’ study, Outbreak-associated Salmonella enterica Serotypes and Food Commodities, United States, 1998- 2008.  Created: 9/9/2013 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 9/9/2013.

  11. Typhaea stercorea (Coleoptera : Mycetophagidae), a carrier of Salmonella enterica serovar Infantis in a Danish broiler house

    DEFF Research Database (Denmark)

    Hald, Birthe; Olsen, A.; Madsen, Mogens

    1998-01-01

    In December 1994, Salmonella enterica serovar Infantis (S. Infantis) was accidentally introduced into a Danish broiler house by stocking an S. Infantis-infected broiler nock of 39,900 day-old chicks. At the time of the study, the infection had persisted through e broiler cycles. Typhaea stercorea...

  12. Dissemination of clonal Salmonella enterica serovar Typhimurium isolates causing salmonellosis in Mauritius

    DEFF Research Database (Denmark)

    Issack, M. I.; Migura, Lourdes Garcia; Ramsamy, Veemala D.;

    2013-01-01

    Salmonella enterica serotype Typhimurium is one of the leading causes of salmonellosis in Mauritius, where it has also been associated with outbreaks of foodborne illness. However, little is known about its molecular epidemiology in the country. This study was therefore undertaken to investigate ...

  13. Draft Genome Sequences of Two Salmonella enterica Strains Isolated from Sprouted Chia and Flax Seed Powders

    Science.gov (United States)

    Ronholm, Jennifer; Petronella, Nicholas

    2016-01-01

    A 2014 foodborne salmonellosis outbreak in Canada and the United States implicated, for the first time, sprouted chia seed powder as the vehicle of transmission. Here, we report the draft whole genome sequences of two Salmonella enterica strains isolated from sprouted powders related to the aforementioned outbreak. PMID:27660774

  14. Effects of Pseudomonas chlororaphis and gaseous chlorine dioxide on the survival of Salmonella enterica on tomatoes

    Science.gov (United States)

    Produce contamination incited by Salmonella enterica serovars on tomatoes and various outbreaks of Salmonellisis have been reported periodically. Post-harvest intervention measures applied to limit produce contamination will improve food and consumer safety. The aim of this reserach was to evaluat...

  15. Multiple roles of putrescine and spermidine in stress resistance and virulence of Salmonella enterica serovar Typhimurium

    DEFF Research Database (Denmark)

    Cartas Espinel, Irene; Guerra, Priscila Regina; Jelsbak, Lotte

    2016-01-01

    Polyamines (putrescine and spermidine) are small-cationic amines ubiquitous in nature and present in most living cells. In recent years they have been linked to virulence of several human pathogens including Shigella spp and Salmonella enterica serovar Typhimurium (S. Typhimurium). Central to S...

  16. Persistence of fluoroquinolone-resistant Salmonella enterica serovar Kentucky from poultry and poultry sources in Nigeria

    DEFF Research Database (Denmark)

    Raufu, Ibrahim A.; Fashae, Kayode; Ameh, James A.;

    2014-01-01

    Introduction: This study investigated the antimicrobial resistance and clonality of Salmonella enterica serotype Kentucky in poultry and poultry sources in Nigeria, and compared the isolates with the clone of S. Kentucky STI98-X1 CIPR using (PFGE) and (MIC). Methodology: Fecal samples from chicke...

  17. Survival, elongation, and elevated tolerance of Salmonella enterica serovar Enteritidis at reduced water activity

    NARCIS (Netherlands)

    Kieboom, J.; Kusumaningrum, H.D.; Tempelaars, M.H.; Hazeleger, W.C.; Abee, T.; Beumer, R.R.

    2006-01-01

    Growing microorganisms on dry surfaces, which results in exposure to low water activity (aw), may change their normal morphology and physiological activity. In this study, the morphological changes and cell viability of Salmonella enterica serovar Enteritidis challenged to low aw were analyzed. The

  18. Extremely Drug-Resistant Salmonella enterica Serovar Senftenberg Infections in Patients in Zambia

    DEFF Research Database (Denmark)

    Hendriksen, Rene S.; Joensen, Katrine Grimstrup; Lukwesa-Musyani, Chileshe;

    2013-01-01

    Two cases of extremely drug-resistant Salmonella enterica serovar Senftenberg isolated from patients in Zambia were investigated by utilizing MIC determinations and whole-genome sequencing. The isolates were resistant to, and harbored genes toward, nine drug classes, including fluoroquinolones...

  19. Complete Genome Sequence of a Myoviridae Bacteriophage Infecting Salmonella enterica Serovar Typhimurium.

    Science.gov (United States)

    Paradiso, Rubina; Orsini, Massimiliano; Bolletti Censi, Sergio; Borriello, Giorgia; Galiero, Giorgio

    2016-01-01

    The bacteriophage 118970_sal3 was isolated from water buffalo feces in southern Italy, exhibiting lytic activity against Salmonella enterica serovar Typhimurium. This bacteriophage belongs to the Myoviridae family and has a 39,464-bp double-stranded DNA (ds-DNA) genome containing 53 coding sequences (CDSs). PMID:27688333

  20. Complete Genome Sequence of a Lytic Siphoviridae Bacteriophage Infecting Several Serovars of Salmonella enterica.

    Science.gov (United States)

    Paradiso, Rubina; Lombardi, Serena; Iodice, Maria Grazia; Riccardi, Marita Georgia; Orsini, Massimiliano; Bolletti Censi, Sergio; Galiero, Giorgio; Borriello, Giorgia

    2016-01-01

    The bacteriophage 100268_sal2 was isolated from water buffalo feces in southern Italy, exhibiting lytic activity against several subspecies of Salmonella enterica This bacteriophage belongs to the Siphoviridae family and has a 125,114-bp double-stranded DNA (ds-DNA) genome containing 188 coding sequences (CDSs). PMID:27688334

  1. Complete Genome Sequence of a Myoviridae Bacteriophage Infecting Salmonella enterica Serovar Typhimurium

    Science.gov (United States)

    Paradiso, Rubina; Orsini, Massimiliano; Bolletti Censi, Sergio; Galiero, Giorgio

    2016-01-01

    The bacteriophage 118970_sal3 was isolated from water buffalo feces in southern Italy, exhibiting lytic activity against Salmonella enterica serovar Typhimurium. This bacteriophage belongs to the Myoviridae family and has a 39,464-bp double-stranded DNA (ds-DNA) genome containing 53 coding sequences (CDSs). PMID:27688333

  2. Complete Genome Sequence of a Lytic Siphoviridae Bacteriophage Infecting Several Serovars of Salmonella enterica

    Science.gov (United States)

    Paradiso, Rubina; Lombardi, Serena; Iodice, Maria Grazia; Riccardi, Marita Georgia; Orsini, Massimiliano; Bolletti Censi, Sergio; Galiero, Giorgio

    2016-01-01

    The bacteriophage 100268_sal2 was isolated from water buffalo feces in southern Italy, exhibiting lytic activity against several subspecies of Salmonella enterica. This bacteriophage belongs to the Siphoviridae family and has a 125,114-bp double-stranded DNA (ds-DNA) genome containing 188 coding sequences (CDSs). PMID:27688334

  3. Subtyping of Salmonella enterica subspecies I using single nucleotide polymorphisms in adenylate cyclase (cyaA)

    Science.gov (United States)

    Methods to rapidly identify serotypes of Salmonella enterica subspecies I are of vital importance for protecting the safety of food. To supplement the serotyping method dkgB-linked intergenic sequence ribotyping (ISR), single nucleotide polymorphisms (SNPs) were characterized within adenylate cyclas...

  4. Draft Genome Sequences of Four Salmonella enterica Strains Isolated from Turkey-Associated Sources

    Science.gov (United States)

    Han, Jing; Gokulan, Kuppan; Zhao, Shaohua; Gies, Allen

    2016-01-01

    We report the draft genomes of four Salmonella enterica isolates evaluated for the contribution of plasmids to virulence. Strains SE163A, SE696A, and SE710A carry plasmids demonstrated to facilitate plasmid-associated virulence, while SE819 is less virulent and has been used as a recipient for conjugation experiments to assess plasmid-encoded virulence mechanisms. PMID:27738037

  5. Draft Genome Sequences of Two Salmonella enterica Strains Isolated from Sprouted Chia and Flax Seed Powders.

    Science.gov (United States)

    Ronholm, Jennifer; Petronella, Nicholas; Tamber, Sandeep

    2016-01-01

    A 2014 foodborne salmonellosis outbreak in Canada and the United States implicated, for the first time, sprouted chia seed powder as the vehicle of transmission. Here, we report the draft whole genome sequences of two Salmonella enterica strains isolated from sprouted powders related to the aforementioned outbreak. PMID:27660774

  6. Prevalence and Characterization of Salmonella enterica and Salmonella Bacteriophages Recovered from Beef Cattle Feedlots in South Texas.

    Science.gov (United States)

    Xie, Yicheng; Savell, Jeffrey W; Arnold, Ashley N; Gehring, Kerri B; Gill, Jason J; Taylor, T Matthew

    2016-08-01

    Asymptomatic Salmonella carriage in beef cattle is a food safety concern, and the beef feedlot environment may function as a reservoir of this pathogen. The goal of this study was to identify and isolate Salmonella and Salmonella bacteriophages from beef cattle feedlot environments in order to better understand the microbial ecology of Salmonella and identify phages that might be useful as anti-Salmonella beef safety interventions. Three feedlots in south Texas were visited, and 27 distinct samples from each source were collected from dropped feces, feed from feed bunks, drinking water from troughs, and soil in cattle pens (n = 108 samples). Preenrichment, selective enrichment, and selective/differential isolation of Salmonella were performed on each sample. A representative subset of presumptive Salmonella isolates was prepared for biochemical identification and serotyping. Samples were pooled by feedlot and sample type to create 36 samples and enriched to recover phages. Recovered phages were tested for host range against two panels of Salmonella hosts. Salmonella bacteria were identified in 20 (18.5%) of 108 samples by biochemical and/or serological testing. The serovars recovered included Salmonella enterica serovars Anatum, Muenchen, Altona, Kralingen, Kentucky, and Montevideo; Salmonella Anatum was the most frequently recovered serotype. Phage-positive samples were distributed evenly over the three feedlots, suggesting that phage prevalence is not strongly correlated with the presence of culturable Salmonella. Phages were found more frequently in soil and feces than in feed and water samples. The recovery of bacteriophages in the Salmonella-free feedlot suggests that phages might play a role in suppressing the Salmonella population in a feedlot environment.

  7. Analysis of Plasmid and Chromosomal DNA of Multidrug-Resistant Salmonella enterica Serovar Typhi from Asia

    Science.gov (United States)

    Mirza, S.; Kariuki, S.; Mamun, K. Z.; Beeching, N. J.; Hart, C. A.

    2000-01-01

    Molecular analysis of chromosomal DNA from 193 multidrug-resistant (MDR) Salmonella enterica serovar Typhi isolates from 1990 to 1995 from Pakistan, Kuwait, Malaysia, Bangladesh, and India produced a total of five major different pulsed-field gel electrophoresis (PFGE) patterns. Even within a particular country MDR S. enterica serovar Typhi DNA was found to be in different PFGE groups. Similar self-transferable 98-MDa plasmids belonging to either incompatibility group incHI1 or incHI1/FIIA were implicated in the MDR phenotype in S. enterica serovar Typhi isolates from all the locations except Quetta, Pakistan, where the majority were of incFIA. A total of five different PFGE genotypes with six different plasmids, based on incompatibility and restriction endonuclease analysis groups, were found among these MDR S. enterica serovar Typhi isolates. PMID:10747124

  8. A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species

    Directory of Open Access Journals (Sweden)

    M. Radhika

    2014-06-01

    Full Text Available Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC. The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae organisms and also against other pathogens. Further evaluation of multiplex PCR was undertaken on 50 natural samples of chicken, eggs and poultry litter and results compared with conventional culture isolation and identification procedure. The multiplex PCR identified the presence of Salmonella and Shigella strains with a short pre-enrichment step of 5 h in peptone water and the same samples were processed by conventional procedures for comparison. Therefore, this reported multiplex PCR can serve as an alternative to the tedious time-consuming procedure of culture and identification in food safety laboratories.

  9. A comparative study of thermal and acid inactivation kinetics in fruit juices of Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Senftenberg grown at acidic conditions.

    Science.gov (United States)

    Alvarez-Ordóñez, Avelino; Fernández, Ana; Bernardo, Ana; López, Mercedes

    2009-11-01

    Acid and heat inactivation in orange and apple juices of Salmonella enterica serovar Typhimurium Colección Española de Cultivos Tipo (i.e., Spanish Type Culture Collection) 443 (CECT 443) (Salmonella Typhimurium) and S. enterica serovar Senftenberg CECT 4384 (Salmonella Senftenberg) grown in buffered brain heart infusion (pH 7.0) and acidified brain heart infusion up to pH 4.5 with acetic, citric, lactic, and hydrochloric acids was evaluated. Acid adaptation induced an adaptive response that increased the subsequent resistance to extreme pH conditions (pH 2.5) and to heat, although the magnitude of these responses differed between the two isolates and fruit juices. The acid resistance in orange juice for acid-adapted cells (D-values of 28.3-34.5 min for Salmonella Senftenberg and 30.0-39.2 min for Salmonella Typhimurium) resulted to be about two to three times higher than that corresponding to non-acid-adapted cells. In apple juice, acid-adapted Salmonella Senftenberg cells survived better than those of Salmonella Typhimurium, obtaining mean D-values of 114.8 +/- 12.3 and 41.9 +/- 2.5 min, respectively. The thermotolerance of non-acid-adapted Salmonella Typhimurium in orange (D(58)-value: 0.028 min) and apple juices (D(58)-value: 0.10 min) was approximately double for acid-adapted cells. This cross-protection to heat was more strongly expressed in Salmonella Senftenberg. D(58)-values obtained for non-acid-adapted cells in orange (0.11 min) and apple juices (0.19 min) increased approximately 10 and 5 times, respectively, after their growth in acidified media. The conditions prevailing during bacterial growth and heat treatment did not significantly influence the z-values observed (6.0 +/- 0.3 degrees C for Salmonella Typhimurium and 7.0 +/- 0.3 degrees C for Salmonella Senftenberg). The enhanced acid resistance found for both isolates could enable them to survive for prolonged time periods in the gastrointestinal tract, increasing the risk of illness. Further, it

  10. Diversity and antimicrobial susceptibility of Salmonella enterica serovars isolated from pig farms in Ibadan, Nigeria

    DEFF Research Database (Denmark)

    Fashae, Kayode; Hendriksen, Rene S.

    2014-01-01

    Animals including food animals play a significant role in the epidemiology of Salmonella enterica. The control requires identification of sources and institution of targeted interventions. This study investigates the diversity of S. enterica serovars, antimicrobial susceptibility, and occurrence...... of plasmid-mediated quinolone resistance (PMQR) genes in pigs in Ibadan, Nigeria. Pooled fresh pen floor fecal samples of pigs collected from 31 pig farms were cultured; the Salmonella isolates were serotyped and their antimicrobial susceptibility was determined. PMQR genes were screened by polymerase chain....... Other PMQR genes were not detected. Pigs constitute an important source of diverse Salmonella serovars in Ibadan. The isolates were more resistant to old antimicrobials with some multiple resistant. Control measures and regulation of antimicrobials are warranted....

  11. Global gene expression of a murein (Braun) lipoprotein mutant of Salmonella enterica serovar Typhimurium by microarray analysis.

    Science.gov (United States)

    Fadl, A A; Galindo, C L; Sha, J; Klimpel, G R; Popov, V L; Chopra, A K

    2006-06-01

    Braun/murein lipoprotein (Lpp) is one of the major outer membrane components of gram-negative enteric bacteria involved in inflammatory responses and septic shock. In previous studies, we reported that two copies of the lipoprotein (lpp) gene (designated as lppA and lppB) existed on the chromosome of Salmonella enterica serovar Typhimurium. Deletion of both lppA and lppB genes rendered Salmonella defective in invasion, motility, induction of cytotoxicity, and production of inflammatory cytokines/chemokines. The lppAB double-knockout (DKO) mutant was attenuated in mice, and animals immunized with this mutant were protected against subsequent challenge with lethal doses of wild-type (wt) S. Typhimurium. To better understand how deletion of the lpp gene might affect Salmonella virulence, we performed global transcriptional profiling of the genes in the wt and the lppAB DKO mutant of S. Typhimurium using microarrays. Our data revealed alterations in the expression of flagellar genes, invasion-associated type III secretion system genes, and transcriptional virulence gene regulators in the lppAB DKO mutant compared to wt S. Typhimurium. These data correlated with the lppAB DKO mutant phenotype and provided possible mechanism(s) of Lpp-associated attenuation in S. Typhimurium. Although these studies were performed in in vitro grown bacteria, our future research will be targeted at global transcriptional profiling of the genes in in vivo grown wt S. Typhimurium and its Lpp mutant.

  12. Diverse distribution of Toxin-Antitoxin II systems in Salmonella enterica serovars

    Science.gov (United States)

    Di Cesare, Andrea; Losasso, Carmen; Barco, Lisa; Eckert, Ester M.; Conficoni, Daniele; Sarasini, Giulia; Corno, Gianluca; Ricci, Antonia

    2016-01-01

    Type II Toxin-Antitoxin systems (TAs), known for their presence in virulent and antibiotic resistant bacterial strains, were recently identified in Salmonella enterica isolates. However, the relationships between the presence of TAs (ccdAB and vapBC) and the epidemiological and genetic features of different non-typhoidal Salmonella serovars are largely unknown, reducing our understanding of the ecological success of different serovars. Salmonella enterica isolates from different sources, belonging to different serovars and epidemiologically unrelated according to ERIC profiles, were investigated for the presence of type II TAs, plasmid content, and antibiotic resistance. The results showed the ubiquitous presence of the vapBC gene in all the investigated Salmonella isolates, but a diverse distribution of ccdAB, which was detected in the most widespread Salmonella serovars, only. Analysis of the plasmid toxin ccdB translated sequence of four selected Salmonella isolates showed the presence of the amino acid substitution R99W, known to impede in vitro the lethal effect of CcdB toxin in the absence of its cognate antitoxin CcdA. These findings suggest a direct role of the TAs in promoting adaptability and persistence of the most prevalent Salmonella serovars, thus implying a wider eco-physiological role for these type II TAs. PMID:27357537

  13. Minimal Functions and Physiological Conditions Required for Growth of Salmonella enterica on Ethanolamine in the Absence of the Metabolosome

    OpenAIRE

    Brinsmade, Shaun R.; Paldon, Tenzin; Escalante-Semerena, Jorge C

    2005-01-01

    During growth on ethanolamine, Salmonella enterica synthesizes a multimolecular structure that mimics the carboxysome used by some photosynthetic bacteria to fix CO2. In S. enterica, this carboxysome-like structure (hereafter referred to as the ethanolamine metabolosome) is thought to contain the enzymatic machinery needed to metabolize ethanolamine into acetyl coenzyme A (acetyl-CoA). Analysis of the growth behavior of mutant strains of S. enterica lacking specific functions encoded by the 1...

  14. Antimicrobial activity of sweet basil and thyme against salmonella enterica serotype Enteritidis in egg-based pasta

    Directory of Open Access Journals (Sweden)

    Stojiljković Jasmina

    2015-01-01

    Full Text Available Salmonella enterica serotype Enteritidis is known as one of the most common pathogenic bacteria causing salmonellosis in humans. Raw materials of animal origin (eggs, chicken meat are frequent vectors that transmit this bacterium. Since eggs are used for the production of pasta, due to insufficient thermal treatment during pasta drying, they can be a potential risk to consumer health. Different essential oils of herbs can be used to reduce present pathogenic microorganisms. This paper compares a decrease in the number of Salmonella enterica serotype Enteritidis (D ATCC 13076 and Salmonella enterica serotype Enteritidis isolated from outbreaks of salmonellosis in egg-based pasta under the influence of thyme and sweet basil essential oils. The results indicate that the utilized oils were more effective against the epidemic strain than the ATCC strain. In addition, thyme oil caused a more significant inhibition of Salmonella enterica serotype Enteritidis during the production process.

  15. Metabolic parameters linked by Phenotype MicroArray to acid resistance profiles of poultry-associated Salmonella enterica

    Science.gov (United States)

    Phenotype microarrays were analyzed for 51 datasets derived from Salmonella enterica. The top 4 serovars associated with poultry products and one associated with turkey, respectively Typhimurium, Enteritidis, Heidelberg, Infantis and Senftenberg, were represented. Datasets were clustered into two ...

  16. Antibiotic resistances in Listeria monocytogenes and Salmonella enterica isolated from foods with animal origin

    Directory of Open Access Journals (Sweden)

    Baltasar Balsalobre Hernández

    2004-12-01

    Full Text Available Extensive use of antibiotics in both human and animal health and in cattle production has generated resistant microorganisms to common antibiotics. Resistances spread caused by human and animal therapeutic is well known, but we know poorly frecuency of resistant bacteria in foods with animal origin and destinated to human consumers. In this paper, sensitivity to nineteen antibiotics was investigated in Listeria monocytogenes and Salmonella enterica strains isolated from foods with animal origin, including fresh meat, hamburgers, fresh sausages, boiled ham and new-laid chicken eggs. The plate diffusion method of Bauer-Kirby was used.Listeria monocytogenes strains showed a very high sensitivity to all antibiotics checked, with the exception of one strain tetracycline resistant. In contrast, Salmonella enterica showed a high frecuency of resistances, in special to tetracycline, streptomycin, nalidixic acid, ticarcillin, ampicillin and chloramphenicol. Moreover, multi-resistance was a common phenomenon. Twenty percent of S. enterica strains were resistant to four or more antibiotics. Frecuency of resistances was higher in 4,5,12:i:-, Hadar, Typhimurium and Virchow serotypes.In conclusion, Salmonella enterica strains isolated from foods with animal origin and destinated to human consumers are usually resistant to several antibiotics. The significance of this observation and its potential health risk must be investigated.

  17. Herd prevalence of Salmonella enterica infections in Danish slaughter pigs determined by microbiological testing

    DEFF Research Database (Denmark)

    Baggesen, Dorte Lau; Wegener, Henrik Caspar; Bager, Flemming;

    1996-01-01

    As a part of a nationwide programme to survey and control salmonella in pig herds, a microbiological survey of 1363 pig herds was performed in Denmark. A total of 13 468 slaughter pigs were examined at slaughter by culture of 5 g of caecal contents. Overall, 30 different serotypes of Salmonella...... the most frequent (49.1%). Salmonella enterica was found in 302 herds (22.2%), S. Typhimurium was found in 61.1% of these. 279 (23.1%) large herds (producing more than 2600 slaughter pigs per year) were found to be salmonella positive compared with 23 (14.7 %) small herds (annual production of 500 to 550...... slaughter pigs). Practical constraints in the study design did not allow for a firm conclusion on the interplay among herd size, geographical location and occurrence of salmonella. In 284 of 302 infected herds (94.0%) only one serotype was detected, Infections with two different serovars were seen in 18...

  18. Resistance to antibiotics and genotype characteristics of Salmonella enterica subspecies enterica serovar Mbandaka isolated from poultry

    OpenAIRE

    Vidanović Dejan; Sabo Z.; Kilibarda Nataša; Živadinović Mira; Žarković Aleksandar; Matović Kazimir

    2008-01-01

    Salmonellas are one of the main zoonotic pathogens whose reservoirs are poultry, cattle and pigs. By means of the food chain salmonellas can be transferred to humans through contaminated food of animal origin. Multiresistant strains Salmonella are particularly dangerous since they can transfer genes of resistance to antibiotics to other microorganisms. Control of salmonellas primarily depends on a good surveillance system and knowledge of the strain types present in the epizootiologic area. I...

  19. Acquisition of Resistance to Extended-Spectrum Cephalosporins by Salmonella enterica subsp. enterica Serovar Newport and Escherichia coli in the Turkey Poult Intestinal Tract

    OpenAIRE

    Poppe, C; Martin, L. C.; Gyles, C. L.; Reid-Smith, R.; Boerlin, P.; McEwen, S A; Prescott, J F; Forward, K R

    2005-01-01

    Salmonella enterica subsp. enterica serovar Newport resistant to the extended-spectrum cephalosporins (ESCs) and other antimicrobials causes septicemic salmonellosis in humans and animals and is increasingly isolated from humans, animals, foods, and environmental sources. Mechanisms whereby serovar Newport bacteria become resistant to ESCs and other classes of antimicrobials while inhabiting the intestinal tract are not well understood. The present study shows that 25.3% of serovar Newport st...

  20. Complete Genome Sequence of Salmonella enterica subsp. enterica Serovar Indiana C629, a Carbapenem-Resistant Bacterium Isolated from Chicken Carcass in China.

    Science.gov (United States)

    Wang, Wei; Liu, Feng; Peng, Zixin; Li, Fengqin; Ma, Aiguo

    2016-01-01

    The carbapenem-resistant Salmonella enterica subsp. enterica serovar Indiana strain C629 was isolated from a chicken carcass collected from a slaughterhouse in Qingdao, China. The complete genome sequence of C629 contains a circular 4,791,723-bp chromosome and a circular 210,106-bp plasmid. Genes involved in carbapenem resistance of this bacterium were identified by whole-genome analysis. PMID:27417837

  1. Complete Genome Sequence of Salmonella enterica subsp. enterica Serovar Indiana C629, a Carbapenem-Resistant Bacterium Isolated from Chicken Carcass in China

    Science.gov (United States)

    Liu, Feng; Peng, Zixin; Li, Fengqin

    2016-01-01

    The carbapenem-resistant Salmonella enterica subsp. enterica serovar Indiana strain C629 was isolated from a chicken carcass collected from a slaughterhouse in Qingdao, China. The complete genome sequence of C629 contains a circular 4,791,723-bp chromosome and a circular 210,106-bp plasmid. Genes involved in carbapenem resistance of this bacterium were identified by whole-genome analysis. PMID:27417837

  2. Occurrence of multidrug-resistant Salmonella enterica serovar Enteritidis isolates from poultry in Iran

    Directory of Open Access Journals (Sweden)

    Ghaderi, R.

    2016-03-01

    Full Text Available Salmonella enterica is recognized as one of the major food-borne pathogens with more than 2,500 serotypes worldwide. The present study addresses antimicrobial resistance of Salmonella enterica serovar Enteritidis isolates in Iran. A collection of 151 Salmonella spp. isolates collected from poultry were serotyped to identify Salmonella Enteritidis. Sixty-one Salmonella Enteritidis were subsequently tested against 30 antimicrobials. A high frequency of antimicrobial resistance was observed against nitrofurantoin (n=55, 90.2% followed by nalidixic acid (n=41, 67.2%, and cephalexin (n=23, 37.7%. Multi-drug resistance were observed in 35 (57.4% out of 61 isolates. Twenty-six antimicrobial resistance patterns were observed among the 61 Salmonella Enteritidis. All isolates were susceptible to ofloxacin, imipenem, enrofloxacin, chloramphenicol, gentamicin, and 3rd and 4th generation cephalosporins. In conclusion, our results revealed that implementing new policies toward overuse of antimicrobial drugs in Iranian poultry industry are of great importance.

  3. Identification of Key Genes in the Response to Salmonella enterica Enteritidis, Salmonella enterica Pullorum, and Poly(I:C) in Chicken Spleen and Caecum

    Science.gov (United States)

    Sheng, Zhongwei; Dai, Aiqin; Zhai, Fei; Li, Jianchao; Xia, Mingxiu; Hua, Dengke; Xu, Lu; Wang, Hongzhi; Chen, Jing; Liu, Lu; Chen, Guohong

    2014-01-01

    Salmonella enterica Enteritidis (S. Enteritidis) and Salmonella enterica Pullorum (S. pullorum) are regarded as a threat to poultry production. This study's aim is to characterize the expression profiles in response to three different challenges and to identify infection-related genes in the chicken spleen and caecum. Groups of the Chinese chicken breed Langshan were challenged with either S. Enteritidis, S. pullorum, or poly(I:C). The concentrations of cytokines and antibodies and the Salmonella colonization level of the caecum and liver were detected in each group at 7 days postinfection. Expression microarray experiments were conducted using mRNA isolated from both spleen and caecum. Crucial differentially expressed genes (DEGs) associated with immunity were identified. Four DEGs were identified in spleen of all three challenge groups (RBM16, FAH, SOX5, and RBM9) and different four genes in caecum (SOUL, FCN2, ANLN, and ACSL1). Expression profiles were clearly different among the three challenged groups. Genes enriched in the spleen of birds infected with S. pullorum were enriched in lymphocyte proliferation related pathways, but the enriched genes in the caecum of the same group were primarily enriched in innate immunity or antibacterial responses. The DEGs that appear across all three challenge groups might represent global response factors for different pathogens. PMID:24707473

  4. Identification of Key Genes in the Response to Salmonella enterica Enteritidis, Salmonella enterica Pullorum, and Poly(I:C in Chicken Spleen and Caecum

    Directory of Open Access Journals (Sweden)

    Teng Ma

    2014-01-01

    Full Text Available Salmonella enterica Enteritidis (S. Enteritidis and Salmonella enterica Pullorum (S. pullorum are regarded as a threat to poultry production. This study’s aim is to characterize the expression profiles in response to three different challenges and to identify infection-related genes in the chicken spleen and caecum. Groups of the Chinese chicken breed Langshan were challenged with either S. Enteritidis, S. pullorum, or poly(I:C. The concentrations of cytokines and antibodies and the Salmonella colonization level of the caecum and liver were detected in each group at 7 days postinfection. Expression microarray experiments were conducted using mRNA isolated from both spleen and caecum. Crucial differentially expressed genes (DEGs associated with immunity were identified. Four DEGs were identified in spleen of all three challenge groups (RBM16, FAH, SOX5, and RBM9 and different four genes in caecum (SOUL, FCN2, ANLN, and ACSL1. Expression profiles were clearly different among the three challenged groups. Genes enriched in the spleen of birds infected with S. pullorum were enriched in lymphocyte proliferation related pathways, but the enriched genes in the caecum of the same group were primarily enriched in innate immunity or antibacterial responses. The DEGs that appear across all three challenge groups might represent global response factors for different pathogens.

  5. Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India

    Institute of Scientific and Technical Information of China (English)

    Irfan; Ahmad; Mir; Sudhir; Kumar; Kashyap; Sunil; Maherchandani

    2015-01-01

    Objective: To study the occurrence and serotype diversity of Salmonella isolates in different species of poultry(chicken, emu and duck) and determine their resistance pattern against various antibiotics of different classes.Methods: About 507 samples comprising 202 caecal contents and 305 fecal samples from chicken, emu and duck were processed for isolation of Salmonella enterica. Salmonellae were isolated and detected by standard protocol of ISO 6579 Amendment 1: Annex D. Genetic confirmation was also made by using 16 S r RNA genus specific PCR. Serotype specific PCR was also done to detect the most common serovars viz. Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Gallinarum. All obtained isolates were subjected to a set of 25 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method.Results: Out of 507 samples processed, 32 isolates of Salmonella enterica(18 from caecal contents and 14 from faecal samples) were obtained, of which 24 belonged to 6 different serovars, 6 were untypeable and 2 were rough strains. Salmonella Enteritidis was the most predominant serotype(9), followed by Salmonella Typhimurium(5), Salmonella Virchow(4), Salmonella Gallinarum(3), Salmonella Reading(2) and Salmonella Altona(1). Antibiotic resistance pattern was maximum(100%) to oxacillin, penicillin and clindamycin, followed by ampicillin(68.75%), tetracycline(65.62%), nalidixic acid(56.25%) and colistin(46.87%). High sensitivity of isolates was recorded for chloramphenicol(96.87%) followed by meropenem(84.37%). Conclusions: Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in prophylaxis

  6. Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India

    Institute of Scientific and Technical Information of China (English)

    Irfan Ahmad Mir; Sudhir Kumar Kashyap; Sunil Maherchandani

    2015-01-01

    Objective:To study the occurrence and serotype diversity of Salmonella isolates in different species of poultry (chicken, emu and duck) and determine their resistance pattern against various antibiotics of different classes. Methods:About 507 samples comprising 202 caecal contents and 305 fecal samples from chicken, emu and duck were processed for isolation of Salmonella enterica. Salmonellae were isolated and detected by standard protocol of ISO 6579 Amendment 1:Annex D. Genetic confirmation was also made by using 16S rRNA genus specific PCR. Serotype specific PCR was also done to detect the most common serovars viz. Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Gallinarum. All obtained isolates were subjected to a set of 25 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method. Results:Out of 507 samples processed, 32 isolates of Salmonella enterica (18 from caecal contents and 14 from faecal samples) were obtained, of which 24 belonged to 6 different serovars, 6 were untypeable and 2 were rough strains. Salmonella Enteritidis was the most predominant serotype (9), followed by Salmonella Typhimurium (5), Salmonella Virchow (4), Salmonella Gallinarum (3), Salmonella Reading (2) and Salmonella Altona (1). Antibiotic resistance pattern was maximum (100%) to oxacillin, penicillin and clindamycin, followed by ampicillin (68.75%), tetracycline (65.62%), nalidixic acid (56.25%) and colistin (46.87%). High sensitivity of isolates was recorded for chloramphenicol (96.87%) followed by meropenem (84.37%). Conclusions:Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in

  7. Repression of Salmonella enterica phoP expression by small molecules from physiological bile.

    Science.gov (United States)

    Antunes, L Caetano M; Wang, Melody; Andersen, Sarah K; Ferreira, Rosana B R; Kappelhoff, Reinhild; Han, Jun; Borchers, Christoph H; Finlay, B Brett

    2012-05-01

    Infection with Salmonella enterica serovar Typhi in humans causes the life-threatening disease typhoid fever. In the laboratory, typhoid fever can be modeled through the inoculation of susceptible mice with Salmonella enterica serovar Typhimurium. Using this murine model, we previously characterized the interactions between Salmonella Typhimurium and host cells in the gallbladder and showed that this pathogen can successfully invade gallbladder epithelial cells and proliferate. Additionally, we showed that Salmonella Typhimurium can use bile phospholipids to grow at high rates. These abilities are likely important for quick colonization of the gallbladder during typhoid fever and further pathogen dissemination through fecal shedding. To further characterize the interactions between Salmonella and the gallbladder environment, we compared the transcriptomes of Salmonella cultures grown in LB broth or physiological murine bile. Our data showed that many genes involved in bacterial central metabolism are affected by bile, with the citric acid cycle being repressed and alternative respiratory systems being activated. Additionally, our study revealed a new aspect of Salmonella interactions with bile through the identification of the global regulator phoP as a bile-responsive gene. Repression of phoP expression could also be achieved using physiological, but not commercial, bovine bile. The biological activity does not involve PhoPQ sensing of a bile component and is not caused by bile acids, the most abundant organic components of bile. Bioactivity-guided purification allowed the identification of a subset of small molecules from bile that can elicit full activity; however, a single compound with phoP inhibitory activity could not be isolated, suggesting that multiple molecules may act in synergy to achieve this effect. Due to the critical role of phoP in Salmonella virulence, further studies in this area will likely reveal aspects of the interaction between Salmonella

  8. Association between indoor environmental contamination by Salmonella enterica and contamination of eggs on layer farms.

    Science.gov (United States)

    Gole, Vaibhav C; Torok, Valeria; Sexton, Margaret; Caraguel, Charles G B; Chousalkar, Kapil K

    2014-09-01

    This study involves longitudinal and point-in-time surveys of Salmonella carriage and environmental contamination on two commercial cage layer farms positive for Salmonella enterica subsp. enterica serovar Typhimurium (flock A age, 32 weeks; flock B age, 34 weeks). Salmonella-positive fecal, egg belt, and dust samples were all unconditionally associated with eggshells testing positive for Salmonella. The odds of an eggshell testing positive for Salmonella were 91.8, 61.5, and 18.2 times higher when fecal, egg belt, and dust samples, respectively, tested positive for Salmonella. The agreement between the culture-based methods and real-time PCR on preenriched broths for detecting Salmonella was almost perfect for eggshell (observed agreement, 99.19%; kappa coefficient, 0.94) and egg belt samples (observed agreement, 95%; kappa coefficient, 0.88), and it was substantial for fecal (observed agreement, 87.14%; kappa coefficient, 0.47) and floor dust samples (observed agreement, 80.61%; kappa coefficient, 0.58). A 1-log increase in the load of Salmonella detected in the fecal, egg belt, and floor dust samples resulted in 35%, 43%, and 45% increases, respectively (P < 0.001), in the odds of an eggshell testing positive for Salmonella. The multilocus variable-number tandem-repeat analysis (MLVA) patterns of the S. Typhimurium strains isolated from flock A were distinct from those of flock B. S. Typhimurium strains detected from human food poisoning cases exhibited an MLVA pattern similar to those of the strains isolated from flocks A and B.

  9. Hatchery-borne Salmonella enterica serovar Tennessee infections in broilers

    DEFF Research Database (Denmark)

    Christensen, J.P.; Brown, D.J.; Madsen, Mogens;

    1997-01-01

    minor genotypic variation. Three different ribotypes were demonstrated when EcoRI was used for digestion of DNA. Two types were obtained by the use of HindIII. Nine different plasmids and seven different plasmid profiles were demonstrated. A 180 kb plasmid was, however, only demonstrated in isolates...... which were protected from routine cleaning and disinfection. Subsequent inclusion of these areas into the sanitation programme resulted in the elimination of S. enterica ser. Tennessee from the hatchers, and a decreasing prevalence of S. enterica ser. Tennessee was observed in broiler flocks during...

  10. PCR Method To Identify Salmonella enterica Serovars Typhi, Paratyphi A, and Paratyphi B among Salmonella Isolates from the Blood of Patients with Clinical Enteric Fever▿

    Science.gov (United States)

    Levy, Haim; Diallo, Souleymane; Tennant, Sharon M.; Livio, Sofie; Sow, Samba O.; Tapia, Milagritos; Fields, Patricia I.; Mikoleit, Matthew; Tamboura, Boubou; Kotloff, Karen L.; Lagos, Rosanna; Nataro, James P.; Galen, James E.; Levine, Myron M.

    2008-01-01

    PCR methodology was developed to identify Salmonella enterica serovars Typhi, Paratyphi A, and Paratyphi B. One multiplex PCR identifies serogroup D, A, and B and Vi-positive strains; another confirms flagellar antigen “d,” “a,” or “b.” Blinded testing of 664 Malian and Chilean Salmonella blood isolates demonstrated 100% sensitivity and specificity. PMID:18367574

  11. Inhibition of Salmonella enterica biofilm formation using small-molecule adenosine mimetics.

    Science.gov (United States)

    Koopman, Jacob A; Marshall, Joanna M; Bhatiya, Aditi; Eguale, Tadesse; Kwiek, Jesse J; Gunn, John S

    2015-01-01

    Biofilms have been widely implicated in chronic infections and environmental persistence of Salmonella enterica, facilitating enhanced colonization of surfaces and increasing the ability of the bacteria to be transmitted to new hosts. Salmonella enterica serovar Typhi biofilm formation on gallstones from humans and mice enhances gallbladder colonization and bacterial shedding, while Salmonella enterica serovar Typhimurium biofilms facilitate long-term persistence in a number of environments important to food, medical, and farming industries. Salmonella regulates expression of many virulence- and biofilm-related processes using kinase-driven pathways. Kinases play pivotal roles in phosphorylation and energy transfer in cellular processes and possess an ATP-binding pocket required for their functions. Many other cellular proteins also require ATP for their activity. Here we test the hypothesis that pharmacological interference with ATP-requiring enzymes utilizing adenosine mimetic compounds would decrease or inhibit bacterial biofilm formation. Through the screening of a 3,000-member ATP mimetic library, we identified a single compound (compound 7955004) capable of significantly reducing biofilm formation by S. Typhimurium and S. Typhi. The compound was not bactericidal or bacteriostatic toward S. Typhimurium or cytotoxic to mammalian cells. An ATP-Sepharose affinity matrix technique was used to discover potential protein-binding targets of the compound and identified GroEL and DeoD. Compound 7955004 was screened against other known biofilm-forming bacterial species and was found to potently inhibit biofilms of Acinetobacter baumannii as well. The identification of a lead compound with biofilm-inhibiting capabilities toward Salmonella provides a potential new avenue of therapeutic intervention against Salmonella biofilm formation, with applicability to biofilms of other bacterial pathogens.

  12. Resident bacteria on leaves enhance survival of immigrant cells of Salmonella enterica.

    Science.gov (United States)

    Poza-Carrion, Cesar; Suslow, Trevor; Lindow, Steven

    2013-04-01

    Although Salmonella enterica apparently has comparatively low epiphytic fitness on plants, external factors that would influence its ability to survive on plants after contamination would be of significance in the epidemiology of human diseases caused by this human pathogen. Viable population sizes of S. enterica applied to plants preinoculated with Pseudomonas syringae or either of two Erwinia herbicola strains was ≥10-fold higher than that on control plants that were not precolonized by such indigenous bacteria when assessed 24 to 72 h after the imposition of desiccation stress. The protective effect of P. fluorescens, which exhibited antibiosis toward S. enterica in vitro, was only ≈50% that conferred by other bacterial strains. Although S. enterica could produce small cellular aggregates after incubation on wet leaves for several days, and the cells in such aggregates were less susceptible to death upon acute dehydration than solitary cells (as determined by propidium iodide staining), most Salmonella cells were found as isolated cells when it was applied to leaves previously colonized by other bacterial species. The proportion of solitary cells of S. enterica coincident with aggregates of cells of preexisting epiphytic species that subsequently were judged as nonviable by viability staining on dry leaves was as much as 10-fold less than those that had landed on uncolonized portions of the leaf. Thus, survival of immigrant cells of S. enterica on plants appears to be strongly context dependent, and the presence of common epiphytic bacteria on plants can protect such immigrants from at least one key stress (i.e., desiccation) encountered on leaf surfaces. PMID:23506362

  13. National surveillance of Salmonella enterica in food-producing animals in Japan

    Directory of Open Access Journals (Sweden)

    Kijima Mayumi

    2009-08-01

    Full Text Available Abstract A total of 518 fecal samples collected from 183 apparently healthy cattle, 180 pigs and 155 broilers throughout Japan in 1999 were examined to determine the prevalence and antimicrobial susceptibility of Salmonella. The isolation rates were 36.1% in broilers, 2.8% in pigs and 0.5% in cattle. S. enterica Infantis was the most frequent isolate, found in 22.6% of broiler fecal samples. Higher resistance rates were observed against oxytetracycline (82.0%, dihydrostreptomycin (77.9%, kanamycin (41.0% and trimethoprim (35.2%. Resistance rates to ampicillin, ceftiofur, bicozamycin, chloramphenicol and nalidixic acid were S. enterica Senftenberg was found in the isolates obtained from one broiler fecal sample. This is the first report of cephalosporin-resistant Salmonella directly isolated from food animal in Japan.

  14. Chicken serologic response to Salmonella enterica serotype Typhimurium assessed by Elisa

    Directory of Open Access Journals (Sweden)

    GH Oliveira

    2006-03-01

    Full Text Available This study evaluated two enzyme-linked immunosorbent assays (ELISA in the detection of chicken serologic response against Salmonella enterica sorotype Typhimurium. The assays have used as detecting antigen the soluble bacterial proteins of a non-flagellated strain of Salmonella Typhimurium (AgTM, and antibody conjugated to peroxidase or alkaline phosphatase. According to the results, optimal dilutions of antigen (concentration 5.49 mg/mL and serum samples in both assays were 1:20,000 and 1:1,000, respectively. In such conditions, the ELISA/AgTM was able to detect serological response to Salmonella Typhimurium. Cross-reactions to Salmonella serotypes Gallinarum and Pullorum were seen, but not with other serotypes such as Enteritidis.

  15. Interaction of Salmonella enterica Serovar Typhimurium with Intestinal Organoids Derived from Human Induced Pluripotent Stem Cells.

    Science.gov (United States)

    Forbester, Jessica L; Goulding, David; Vallier, Ludovic; Hannan, Nicholas; Hale, Christine; Pickard, Derek; Mukhopadhyay, Subhankar; Dougan, Gordon

    2015-07-01

    The intestinal mucosa forms the first line of defense against infections mediated by enteric pathogens such as salmonellae. Here we exploited intestinal "organoids" (iHOs) generated from human induced pluripotent stem cells (hIPSCs) to explore the interaction of Salmonella enterica serovar Typhimurium with iHOs. Imaging and RNA sequencing were used to analyze these interactions, and clear changes in transcriptional signatures were detected, including altered patterns of cytokine expression after the exposure of iHOs to bacteria. S. Typhimurium microinjected into the lumen of iHOs was able to invade the epithelial barrier, with many bacteria residing within Salmonella-containing vacuoles. An S. Typhimurium invA mutant defective in the Salmonella pathogenicity island 1 invasion apparatus was less capable of invading the iHO epithelium. Hence, we provide evidence that hIPSC-derived organoids are a promising model of the intestinal epithelium for assessing interactions with enteric pathogens.

  16. The Base Excision Repair system of Salmonella enterica serovar typhimurium counteracts DNA damage by host nitric oxide.

    Directory of Open Access Journals (Sweden)

    Anthony R Richardson

    2009-05-01

    Full Text Available Intracellular pathogens must withstand nitric oxide (NO. generated by host phagocytes. Salmonella enterica serovar Typhimurium interferes with intracellular trafficking of inducible nitric oxide synthase (iNOS and possesses multiple systems to detoxify NO.. Consequently, the level of NO. stress encountered by S. Typhimurium during infection in vivo has been unknown. The Base Excision Repair (BER system recognizes and repairs damaged DNA bases including cytosine and guanine residues modified by reactive nitrogen species. Apurinic/apyrimidinic (AP sites generated by BER glycosylases require subsequent processing by AP endonucleases. S. Typhimurium xth nfo mutants lacking AP endonuclease activity exhibit increased NO. sensitivity resulting from chromosomal fragmentation at unprocessed AP sites. BER mutant strains were thus used to probe the nature and extent of nitrosative damage sustained by intracellular bacteria during infection. Here we show that an xth nfo S. Typhimurium mutant is attenuated for virulence in C3H/HeN mice, and virulence can be completely restored by the iNOS inhibitor L-NIL. Inactivation of the ung or fpg glycosylase genes partially restores virulence to xth nfo mutant S. Typhimurium, demonstrating that NO. fluxes in vivo are sufficient to modify cytosine and guanine bases, respectively. Mutants lacking ung or fpg exhibit NO.-dependent hypermutability during infection, underscoring the importance of BER in protecting Salmonella from the genotoxic effects of host NO.. These observations demonstrate that host-derived NO. damages Salmonella DNA in vivo, and the BER system is required to maintain bacterial genomic integrity.

  17. Genetic and molecular analysis of the virulence plasmid of Salmonella enterica serovar Typhimurium

    OpenAIRE

    García Quintanilla, Meritxell de Jesús

    2010-01-01

    Salmonella enterica serovar Typhimurium posee un plásmido de gran tamaño (93 Kb) llamado plásmido de virulencia o pSLT. Dicho plásmido contiene el locus spv, esencial para la infección sistémica en roedores. El plásmido fue presumiblemente adquirido por el hospedador ancestral de Salmonella mediante conjugación. En la mayoría de las estirpes del serovar Typhimurium, el plásmido pSLT contiene un operón tra completo. En esta tesis se han realizado diversos análisis estructurales y funcionales e...

  18. Thioridazine protects the mouse from a virulent infection by Salmonella enterica serovar Typhimurium 74

    DEFF Research Database (Denmark)

    Dasgupta, Asish; Mukherjee, Sayanti; Chaki, Shaswati;

    2010-01-01

    When administered to mice at doses of 100microg/mouse and 200microg/mouse, thioridazine (TDZ) significantly protected animals from the lethality produced by a virulent strain of Salmonella enterica serovar Typhimurium and reduced the number of bacteria retrieved from the spleen, liver and heart...... blood. The protection conferred by TDZ against a virulent Salmonella infection is hypothesised to be due to a reduction in the 55kDa virulence protein of the outer membrane of the organism, as this protein is almost totally absent when the organism is exposed to the phenothiazine. It is further...

  19. Regulation of Salmonella enterica Serovar Typhimurium Invasion Genes by csrA

    OpenAIRE

    Altier, Craig; Suyemoto, Mitsu; Lawhon, Sara D.

    2000-01-01

    Penetration of intestinal epithelial cells by Salmonella enterica serovar Typhimurium requires the expression of invasion genes, found in Salmonella pathogenicity island 1 (SPI1), that encode components of a type III secretion apparatus. These genes are controlled in a complex manner by regulators within SPI1, including HilA and InvF, and those outside SPI1, such as the two-component regulators PhoP/PhoQ and BarA/SirA. We report here that epithelial cell invasion requires the serovar Typhimur...

  20. Population dynamics of Salmonella enterica serotypes in commercial egg and poultry production.

    Science.gov (United States)

    Foley, Steven L; Nayak, Rajesh; Hanning, Irene B; Johnson, Timothy J; Han, Jing; Ricke, Steven C

    2011-07-01

    Fresh and processed poultry have been frequently implicated in cases of human salmonellosis. Furthermore, increased consumption of meat and poultry has increased the potential for exposure to Salmonella enterica. While advances have been made in reducing the prevalence and frequency of Salmonella contamination in processed poultry, there is mounting pressure on commercial growers to prevent and/or eliminate these human pathogens in preharvest production facilities. Several factors contribute to Salmonella colonization in commercial poultry, including the serovar and the infectious dose. In the early 1900s, Salmonella enterica serovars Pullorum and Gallinarum caused widespread diseases in poultry, but vaccination and other voluntary programs helped eradicate pullorum disease and fowl typhoid from commercial flocks. However, the niche created by the eradication of these serovars was likely filled by S. Enteritidis, which proliferated in the bird populations. While this pathogen remains a significant problem in commercial egg and poultry production, its prevalence among poultry has been declining since the 1990s. Coinciding with the decrease of S. Enteritidis, S. Heidelberg and S. Kentucky have emerged as the predominant serovars in commercial broilers. In this review, we have highlighted bacterial genetic and host-related factors that may contribute to such shifts in Salmonella populations in commercial poultry and intervention strategies that could limit their colonization.

  1. First Report of Extended-Spectrum-β-Lactamase-Producing Salmonella enterica Serovar Kentucky Isolated from Poultry in Ireland▿

    OpenAIRE

    Boyle, F; D. Morris; O'Connor, J; DeLappe, N.; Ward, J.; Cormican, M.

    2009-01-01

    Therapy of invasive human salmonellosis is complicated by increasing antimicrobial resistance. Food animals are the principal source of infection with nontyphoid Salmonella. We report the emergence of broad-spectrum-cephalosporin resistance in Salmonella enterica serovar Kentucky in poultry in Ireland.

  2. High-throughput Molecular Determination of Salmonella enterica Serovars Use of Multiplex PCR and Capillary Electrophoresis Analysis

    Science.gov (United States)

    Salmonella enterica is a zoonotic pathogen and is a leading cause of food-borne illness worldwide. There are over 2,500 serotypes of Salmonella reported. Identification of the serotype is key in defining the etiological agent during an outbreak investigation. In the current study, a high-throughput ...

  3. Regulation of fucose and 1,2-propanediol utilization by Salmonella enterica serovar Typhimurium

    OpenAIRE

    Staib, Lena; Fuchs, Thilo M.

    2015-01-01

    After ingestion, Salmonella enterica serovar Typhimurium (S. Typhimurium) encounters a densely populated, competitive environment in the gastrointestinal tract. To escape nutrient limitation caused by the intestinal microbiota, this pathogen has acquired specific metabolic traits to use compounds that are not metabolized by the commensal bacteria. For example, the utilization of 1,2-propanediol (1,2-PD), a product of the fermentation of L-fucose, which is present in foods of herbal origin and...

  4. Antibiotic resistances in Listeria monocytogenes and Salmonella enterica isolated from foods with animal origin

    OpenAIRE

    Baltasar Balsalobre Hernández; Joaquín Hernández-Godoy

    2004-01-01

    Extensive use of antibiotics in both human and animal health and in cattle production has generated resistant microorganisms to common antibiotics. Resistances spread caused by human and animal therapeutic is well known, but we know poorly frecuency of resistant bacteria in foods with animal origin and destinated to human consumers. In this paper, sensitivity to nineteen antibiotics was investigated in Listeria monocytogenes and Salmonella enterica strains isolated from foods with animal orig...

  5. Kinetics of the Natural, Humoral Immune Response to Salmonella enterica Serovar Typhi in Kathmandu, Nepal▿

    OpenAIRE

    Pulickal, Anoop S.; Gautam, Samir; Elizabeth A Clutterbuck; Thorson, Stephen; Basynat, Buddha; Adhikari, Neelam; Makepeace, Katherine; Rijpkema, Sjoerd; Borrow, Ray; Farrar, Jeremy J.; Andrew J Pollard

    2009-01-01

    Typhoid fever is a major public health problem in developing countries, conservatively estimated to occur in 17 million cases and be responsible for 200,000 deaths annually. We investigated the acquisition of natural immunity to Salmonella enterica serovar Typhi in a region where typhoid is endemic by testing sera from an age-stratified sample of 210 healthy participants in Kathmandu, Nepal, for bactericidal activity toward S. Typhi and for anti-Vi capsular polysaccharide antibodies. Bacteric...

  6. Global Genomic Epidemiology of Salmonella enterica Serovar Typhimurium DT104

    OpenAIRE

    Leekitcharoenphon, Pimlapas; Hendriksen, Rene S.; Le Hello, Simon; Weill, François-Xavier; Baggesen, Dorte Lau; Jun, Se-Ran; Ussery, David; Lund, Ole; Crook, Derrick W.; Wilson, Daniel J.; Aarestrup, Frank Møller

    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 structured sequence analysis within a Bayesian framework to reconstruct temporal and spatial phylogenetic trees and estimate the rates of mutation and divergence times of 315S Typhimurium DT104 isolates sa...

  7. Influence of Salmonella enterica serovar Enteritidis infection on the composition of chicken cecal microbiota

    OpenAIRE

    Videnska, Petra; Sisak, Frantisek; Havlickova, Hana; Faldynova, Marcela; Rychlik, Ivan

    2013-01-01

    Background Infection of newly hatched chicks with Salmonella enterica serovar Enteritidis (S. Enteritidis) results in an inflammatory response in the intestinal tract which may influence the composition of gut microbiota. In this study we were therefore interested whether S. Enteritidis induced inflammation results in changes in the cecal microbiota. To reach this aim, we compared the cecal microbiota of non-infected chickens and those infected by S. Enteritidis by pyrosequencing the V3/V4 va...

  8. The STM4195 Gene Product (PanS) Transports Coenzyme A Precursors in Salmonella enterica

    OpenAIRE

    Ernst, Dustin C.; Downs, Diana M.

    2015-01-01

    Coenzyme A (CoA) is a ubiquitous coenzyme involved in fundamental metabolic processes. CoA is synthesized from pantothenic acid by a pathway that is largely conserved among bacteria and eukaryotes and consists of five enzymatic steps. While higher organisms, including humans, must scavenge pantothenate from the environment, most bacteria and plants are capable of de novo pantothenate biosynthesis. In Salmonella enterica, precursors to pantothenate can be salvaged, but subsequent intermediates...

  9. Antioxidant oils and Salmonella enterica Typhimurium reduce tumor in an experimental model of hepatic metastasis

    OpenAIRE

    Sorenson, Brent

    2011-01-01

    Brent S Sorenson, Kaysie L Banton, Lance B Augustin, Arnold S Leonard, Daniel A SaltzmanDepartment of Surgery, University of Minnesota Medical School, Minneapolis, MN, USAAbstract: Fruit seeds high in antioxidants have been shown to have anticancer properties and enhance host protection against microbial infection. Recently we showed that a single oral dose of Salmonella enterica serovar Typhimurium expressing a truncated human interleukin-2 gene (SalpIL2) is avirulent, immunogenic, and reduc...

  10. Accuracy and Sensitivity of Commercial PCR-Based Methods for Detection of Salmonella enterica in Feed ▿

    OpenAIRE

    Koyuncu, Sevinc; Andersson, M. Gunnar; Häggblom, Per

    2010-01-01

    The present study compared the performance of commercial PCR-based Salmonella enterica detection methods (BAX System Q7, the iQ-Check Salmonella II kit, and the TaqMan Salmonella enterica detection kit) with culture-based methods (modified semisolid Rappaport-Vassiliadis [MSRV] and NMKL71) in spiked and naturally contaminated samples of feed mill scrapings (FMS), palm kernel meal (PKM), pelleted feed (PF), rape seed meal (RSM), soybean meal (SM), and wheat grain (WG). When results from the va...

  11. Salmonella Enterica Prevalence in Finishing Pigs at Slaughter Plants in Northern Italy

    Science.gov (United States)

    Bacci, Cristina; Lanzoni, Elisa; Brindani, Franco; Bonardi, Silvia

    2014-01-01

    Finishing pigs carrying Salmonella enterica are believed to be the main source of carcass contamination at the beginning of slaughtering. The aim of this study was to assess the S. enterica carrier status of finishing pigs at herd level by sampling pooled faeces on farm and mesenteric lymph nodes at slaughter in the North East of Italy. Environmental faecal samples belonging to 30 batches of pigs were collected on farm. At slaughter, mesenteric lymph nodes were collected from five randomly selected pigs per batch. S. enterica was isolated from 16 lymph nodes out of 150 (10.6%) and from seven out of 30 (23.3%) faecal samples. Four batches (13.3%) were positive to S. enterica both in lymph nodes and in faeces. The number of batches positive to S. enterica either in lymph nodes or in faeces was 13 out of 30 (43.3%). The most prevalent serovars from lymph nodes were S. Derby (25.0%) and S. Typhimurium monophasic variant 1, 4,[5],12:i:- (18.6%), which were also isolated from faecal material (14.3 and 42.8% respectively). Contaminated faecal material or lymph nodes could be a primary source of carcass contamination at slaughter during evisceration. S. enterica contamination is widespread on pig farms and carrier pigs pass undetected the inspection visits at slaughter, entering the food chain. Therefore, in order to control S. enterica in pigs, the need to quantify possible risk factors at slaughter and develop effective management strategies on farm is of paramount importance to ensure food safety.

  12. Oral immunization of broiler chickens against necrotic enteritis with an attenuated Salmonella vaccine vector expressing Clostridium perfringens antigens.

    Science.gov (United States)

    Kulkarni, R R; Parreira, V R; Sharif, S; Prescott, J F

    2008-08-01

    Necrotic enteritis (NE) in broiler chickens is caused by Clostridium perfringens but currently no effective vaccine is available. Our previous study showed that certain C. perfringens secreted proteins when administered intramuscularly protected chickens against experimental infection. In the current study, genes encoding three C. perfringens proteins: fructose-biphosphate-aldolase (FBA), pyruvate:ferredoxin-oxidoreductase (PFOR) and hypothetical protein (HP), were cloned into an avirulent Salmonella enterica sv. typhimurium vaccine vector. Broiler chickens immunized orally with recombinant Salmonella expressing FBA or HP proteins were significantly protected against NE challenge. Immunized birds developed serum and mucosal antibodies to both clostridial and Salmonella antigens. This study showed the oral immunizing ability of two C. perfringens antigens against NE in broiler chickens through an attenuated Salmonella vaccine vector. PMID:18597901

  13. Shoot Injury Increases the Level of Persistence of Salmonella enterica Serovar Sofia and Listeria innocua on Cos Lettuce and of Salmonella enterica Serovar Sofia on Chive.

    Science.gov (United States)

    Harapas, Dean; Premier, Robert; Tomkins, Bruce; Hepworth, Graham; Ajlouni, Said

    2015-12-01

    Minor shoot injury significantly (P < 0.05) increased the level at which Salmonella enterica serovar Sofia persisted on cos lettuce in the greenhouse. Initial mean counts of the Salmonella on the injured and uninjured cos lettuce were on the order of 6 log CFU/g. After 3 days, the mean count decreased to 4.8 log CFU/g on the injured plants compared with the significantly (P < 0.05) smaller count of 3.4 log CFU/g on the uninjured plants. By the end of the 3-week experiment, the count from the injured plants was 2.9 log CFU/g compared with a count of below the level of detection from the uninjured plants. A similar pattern of bacterial persistence was observed on injured versus uninjured plants by using Listeria innocua on cos lettuce and S. enterica serovar Sofia on chive. The findings reaffirm earlier results with Escherichia coli and increase the impetus to avoid shoot injury during the production of cos lettuce and chive, if bacteria of food safety concern are present.

  14. The Study of Genetic Relationship Among Third Generation Cephalosporin-resistant Salmonella enterica Strains by ERIC-PCR

    OpenAIRE

    Ranjbar, Reza; Naghoni, Ali; Yousefi, Soheila; Ahmadi, Ali; Jonaidi, Nematollah; Panahi, Yunes

    2013-01-01

    Background and Objectives: Salmonella is an important food-borne pathogen responsible for disease in humans and animals. The aim of this study was to investigate the genetic relationship among third generation cephalosporin-resistant Salmonella enterica strains by Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR. Methods: The study included all Salmonella isolates obtained from clinical cases in a pediatric hospital in Tehran, Iran during 2006 to 2009. Antimicrobial susceptibility t...

  15. Variable abattoir conditions affect Salmonella enterica prevalence and meat quality in swine and pork.

    Science.gov (United States)

    Hurd, H S; Gailey, J K; McKean, J D; Griffith, R W

    2005-01-01

    Research suggests that abattoir holding pens pose significant Salmonella enterica risk to swine immediately preharvest. The goal of this study was to evaluate those factors related to holding that increased the prevalence of S. enterica in swine at slaughter. To accomplish this goal, we focused on holding time and flooring. Our objectives were to (1) compare Salmonella enterica prevalence among pigs held for short (15-45 min) versus long (up to 4 h) periods before slaughter; and (2) determine the impact of flooring (slatted vs. concrete) as it relates to the prevalence of S. enterica. The study consisted of seven repetitions at a large volume (11,000 head/day) Midwest abattoir. Each repetition consisted of one truck load of pigs (n = 170) sorted into one of three groups: (1) animals held for a short time (15-45 min) on solid floors (short-hold); (2) animals held for 4 +/- 0.5 h on slatted floors; and (3) animals held for 4 +/- 0.5 h on solid concrete floors. At slaughter, samples were collected from 30 pigs in each group. Cecal contents (20 mL), feces (20 g), and the ileocecal lymph node were cultured for S. enterica. Additionally, the effect of holding time on meat quality parameters (loin pH at 35 min and 6 h, color, drip loss) was evaluated for the first four replicates. The proportion of S. enterica-positive samples was highest (p concrete floors (72.4%), and slightly less for pigs held on slatted floors (63.3%). Animals held for less than 45 min before slaughter demonstrated the lowest proportion of S. enterica-positive samples (52.9%). The pig prevalence, as measured by any one of the three samples being positive, was significantly different (p floors (81%) and those animals held for 45 min or less before slaughter (69%). Meat quality, as measured by multiple parameters, was adversely affected by lack of a rest period. The mean 24-h pH was significantly lower for the short-hold group compared to the other two groups. The mean Minolta L and the drip loss were

  16. Dietary rice bran promotes resistance to Salmonella enterica serovar Typhimurium colonization in mice

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    Kumar Ajay

    2012-07-01

    Full Text Available Abstract Background Dietary rice bran consists of many bioactive components with disease fighting properties; including the capacity to modulate the gut microbiota. Studies point to the important roles of the gut microbiota and the mucosal epithelium in the establishment of protection against enteric pathogens, such as Salmonella. The ability of rice bran to reduce the susceptibility of mice to a Salmonella infection has not been previously investigated. Therefore, we hypothesized that the incorporation of rice bran into the diet would inhibit the colonization of Salmonella in mice through the induction of protective mucosal responses. Results Mice were fed diets containing 0%, 10% and 20% rice bran for one week prior to being orally infected with Salmonella enterica serovar Typhimurium. We found that mice consuming the 10 and 20% rice bran diets exhibited a reduction in Salmonella fecal shedding for up to nine days post-infection as compared to control diet fed animals (p Lactobacillus spp. in rice bran fed mice (p Salmonella entry into mouse small intestinal epithelial cells. Conclusions Increasing rice bran consumption represents a novel dietary means for reducing susceptibility to enteric infection with Salmonella and potentially via induction of native Lactobacillus spp.

  17. Immunoprotectivity of Salmonella enterica serovar Enteritidis virulence protein, InvH, against Salmonella typhi

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    Behzad Dehghani

    2014-08-01

    Full Text Available Objective(s:Typhoid fever is a dreadful disease of a major threat to public health in developing countries. Vaccination with bacterial immunodominant components such as surface proteins may prove as a potent alternative to live attenuated vaccines. InvH, an important part of needle complex in type three secretion system (TTSS plays important role in efficient bacterial adherence and entry into epithelial cells. Materials and Methods:In this work we used a 15 kDa recombinant InvH protein of Salmonella enteric serovar Enteritidis to provoke antibody production in mouse. The mice were immunized by recombinant InvH and challenged with Salmonella typhi. Histopathology of spleen and liver were studied. Results:The immunized mice showed a significant rise of antibody after the second booster. The immunization induced protection against high doses of S. typhi. The bacterial challenge with sera showed significant protection against challenge dose of 2×109 CFU. Immunized sera reacted with          S. typhi markedly. Immunoreaction of bacterially infected sera and InvH protein was significantly higher than the control group. Bacterial loads of S. typhi in spleen was more than liver. Decreased bacterial load was evident in immunized mice after 7 days. Histological examination of the liver showed the immunized mice liver remained unaffected. Conclusion: Efficacy of the virulence protein, InvH, in inhibition of this phenomenon by active immunization was shown here. It may be concluded that InvH, as an antigen, can develop protection against S. typhi infections. InvH may be exploited in protective measures as well as a diagnostic tool in Salmonella infections.

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

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    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.

  19. Emergence of Ciprofloxacin-Resistant Salmonella enterica Serovar Typhi in Italy.

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    Aurora García-Fernández

    Full Text Available In developed countries, typhoid fever is often associated with persons who travel to endemic areas or immigrate from them. Typhoid fever is a systemic infection caused by Salmonella enterica serovar Typhi. Because of the emergence of antimicrobial resistance to standard first-line drugs, fluoroquinolones are the drugs of choice. Resistance to ciprofloxacin by this Salmonella serovar represents an emerging public health issue. Two S. enterica ser. Typhi strains resistant to ciprofloxacin (CIP were reported to the Italian surveillance system for foodborne and waterborne diseases (EnterNet-Italia in 2013. The strains were isolated from two Italian tourists upon their arrival from India. A retrospective analysis of 17 other S. enterica ser. Typhi strains isolated in Italy during 2011-2013 was performed to determine their resistance to CIP. For this purpose, we assayed for susceptibility to antimicrobial agents and conducted PCR and nucleotide sequence analyses. Moreover, all strains were typed using pulsed-field gel electrophoresis to evaluate possible clonal relationships. Sixty-eight percent of the S. enterica ser. Typhi strains were resistant to CIP (MICs, 0.125-16 mg/L, and all isolates were negative for determinants of plasmid-mediated quinolone resistance. Analysis of sequences encoding DNA gyrase and topoisomerase IV subunits revealed mutations in gyrA, gyrB, and parC. Thirteen different clonal groups were detected, and the two CIP-resistant strains isolated from the individuals who visited India exhibited the same PFGE pattern. Because of these findings, the emergence of CIP-resistant S. enterica ser. Typhi isolates in Italy deserves attention, and monitoring antibiotic susceptibility is important for efficiently managing cases of typhoid fever.

  20. Contribution of proton-translocating proteins to the virulence of Salmonella enterica serovars Typhimurium, Gallinarum, and Dublin in chickens and mice.

    Science.gov (United States)

    Turner, A K; Barber, L Z; Wigley, P; Muhammad, S; Jones, M A; Lovell, M A; Hulme, S; Barrow, P A

    2003-06-01

    We investigated the attenuating effects of a range of respiratory chain mutations in three Salmonella serovars which might be used in the development of live vaccines. We tested mutations in nuoG, cydA, cyoA, atpB, and atpH in three serovars of Salmonella enterica: Typhimurium, Dublin, and Gallinarum. All three serovars were assessed for attenuation in their relevant virulence assays of typhoid-like infections. Serovar Typhimurium was assessed in 1-day-old chickens and the mouse. Serovar Gallinarum 9 was assessed in 3-week-old chickens, and serovar Dublin was assessed in 6-week-old mice. Our data show variation in attenuation for the nuoG, cydA, and cyoA mutations within the different serovar-host combinations. However, mutations in atpB and atpH were highly attenuating for all three serovars in the various virulence assays. Further investigation of the mutations in the atp operon showed that the bacteria were less invasive in vivo, showing reduced in vitro survival within phagocytic cells and reduced acid tolerance. We present data showing that this reduced acid tolerance is due to an inability to adapt to conditions rather than a general sensitivity to reduced pH. The data support the targeting of respiratory components for the production of live vaccines and suggest that mutations in the atp operon provide suitable candidates for broad-spectrum attenuation of a range of Salmonella serovars.

  1. Diagnostic Accuracy of Rectoanal Mucosal Swab of Feedlot Cattle for Detection and Enumeration of Salmonella enterica.

    Science.gov (United States)

    Agga, Getahun E; Arthur, Terrance M; Schmidt, John W; Wang, Rong; Brichta-Harhay, Dayna M

    2016-04-01

    Cattle are noted carriers of the foodborne pathogen Salmonella enterica. The perceived need to decrease the potential human health risk posed by excretion of this pathogen has resulted in numerous studies examining the factors that influence Salmonella shedding in cattle. Fecal grab (FG) samples have been the predominant method used to identify cattle colonized or infected with Salmonella; however, FG sampling can be impractical in certain situations, and rectoanal mucosal swabs (RAMS) are a more convenient sample type to collect. Despite a lack of studies comparing FG and RAMS for the detection and enumeration of Salmonella fecal shedding, RAMS is perceived as less sensitive because a smaller amount of feces is cultured. In a cross-sectional study to address these concerns, paired RAMS and FG samples were collected from 403 adult feedlot cattle approximately 90 days prior to harvest. Samples were processed for Salmonella enumeration (direct plating) and detection (enrichment and immunomagnetic separation). In all, 89.6% of RAMS and 98.8% of FG samples were positive for Salmonella, and concordant prevalence outcomes were observed for 90.8% of samples. Mean enumeration values were 3.01 and 3.12 log CFU/ml for RAMS and FG, respectively. The sensitivity and specificity of RAMS were 91% (95% confidence interval [CI]: 87.5 to 93%) and 100% (95% CI: 48 to 100%), respectively, for Salmonella detection. Furthermore, RAMS Salmonella enumeration was substantially concordant (ρc = 0.89; 95% CI: 0.86 to 0.91) with FG values. We conclude that RAMS are a reliable alternative to FG for assessing cattle Salmonella fecal shedding status, especially for cattle shedding high levels of Salmonella. PMID:27052855

  2. Development and comparison of a generic multiple-locus variable-number tandem repeat analysis with PFGE for typing of Salmonella entericasubsp. enterica

    DEFF Research Database (Denmark)

    Kjeldsen, Marianne Kirstine; Torpdahl, Mia; Pedersen, Karl;

    2015-01-01

    Aims Salmonella enterica subsp. enterica causes salmonellosis in humans and animals. Serovar specific multiple-locus variable-number tandem repeat analysis (MLVA) is widely used for Salmonella surveillance; however, isolates have to be serotyped prior to MLVA typing and only the most common...

  3. Effects of P22 bacteriophage on salmonella Enterica subsp. enterica serovar Typhimurium DMC4 strain biofilm formation and eradication

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    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.

  4. Molecular epidemiological characteristics of Salmonella enterica serovars Enteritidis, Typhimurium and Livingstone strains isolated in a Tunisian university hospital.

    Science.gov (United States)

    Ktari, Sonia; Ksibi, Boutheina; Gharsallah, Houda; Mnif, Basma; Maalej, Sonda; Rhimi, Fouzia; Hammami, Adnene

    2016-03-01

    Enteritidis, Typhimurium and Livingstone are the main Salmonella enterica serovars recovered in Tunisia. Here, we aimed to assess the genetic diversity of fifty-seven Salmonella enterica strains from different sampling periods, origins and settings using pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST) and multi-locus variable-number tandem repeat analysis (MLVA). Salmonella Enteritidis, isolated from human and food sources from two regions in Sfax in 2007, were grouped into one cluster using PFGE. However, using MLVA these strains were divided into two clusters. Salmonella Typhimurium strains, recovered in 2012 and represent sporadic cases of human clinical isolates, were included in one PFGE cluster. Nevertheless, the MLVA technique, divided Salmonella Typhimurium isolates into six clusters with diversity index reaching (DI = 0.757). For Salmonella Livingstone which was responsible of two nosocomial outbreaks during 2000-2003, the PFGE and MLVA methods showed that these strains were genetically closely related. Salmonella Enteritidis and Salmonella Livingstone populations showed a single ST lineage ST11 and ST543 respectively. For Salmonella Typhimurium, two MLST sequence types ST19 and ST328 were defined. Salmonella Enteritidis and Salmonella Typhimurium strains were clearly differentiated by MLVA which was not the case using PFGE.

  5. Survival of Salmonella enterica in Dried Turkey Manure and Persistence on Spinach Leaves.

    Science.gov (United States)

    Oni, Ruth A; Sharma, Manan; Buchanan, Robert L

    2015-10-01

    Concerns about the microbiological safety of fresh produce have attracted attention in the past three decades due to multiple foodborne outbreaks. Animal manure contaminated with enteric pathogens has been identified as an important preharvest pathogen source. This study investigated the survival of Salmonella enterica in dust particles of dehydrated turkey manure and how association with manure dust may enhance the survival of salmonellae on leafy greens in the field. The survival of a cocktail of multiple Salmonella serotypes in the dried fecal material of various particle sizes (125 to 500 μm) was examined at varying moisture contents (5, 10, and 15%). Survival times of the pathogen were inversely related to moisture content and particle size of manure dust, with viable Salmonella still detectable for up to 291 days in the smallest particle size (125 μm) with 5% moisture. Association with manure dust particles increased the survival of Salmonella when subjected to UV light both under laboratory conditions and on the surface of spinach leaves in a greenhouse setting. The results of this study suggest that aerosolized manure particles could be a potential vehicle for Salmonella dispersal to leafy greens if the microorganism is present in the dry manure.

  6. A Predominant Multidrug-Resistant Salmonella enterica Serovar Saintpaul Clonal Line in German Turkey and Related Food Products▿

    OpenAIRE

    Beutlich, Janine; Rodríguez, Irene; Schroeter, Andreas; Käsbohrer, Annemarie; Helmuth, Reiner; Guerra, Beatriz

    2010-01-01

    Recently, Salmonella enterica subsp. enterica serovar Saintpaul has increasingly been observed in several countries, including Germany. However, the pathogenic potential and epidemiology of this serovar are not very well known. This study describes biological attributes of S. Saintpaul isolates obtained from turkeys in Germany based on characterization of their pheno- and genotypic properties. Fifty-five S. Saintpaul isolates from German turkeys and turkey-derived food products isolated from ...

  7. High-Throughput Molecular Determination of Salmonella enterica Serovars by Use of Multiplex PCR and Capillary Electrophoresis Analysis▿

    OpenAIRE

    Leader, Brandon T.; Frye, Jonathan G.; Hu, Jinxin; Paula J Fedorka-Cray; Boyle, David S

    2009-01-01

    Salmonella enterica is a leading cause of food-borne illness worldwide and is also a major cause of morbidity and mortality in domestic and wild animals. In the current study, a high-throughput molecular assay was developed to determine the most common clinical and nonhuman serovars of S. enterica in the United States. Sixteen genomic targets were identified based on their differential distribution among common serovars. Primers were designed to amplify regions of each of these targets in a s...

  8. Stressors Influence on Salmonella enterica Serovar Enteritidis Colonization in Broilers

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    E. S. Soliman

    2009-01-01

    Full Text Available Problem statement: Poultry industry usually exposing birds to a variety of actions and stressors includs fasting for gastrointestinal emptying before transportation and where birds are often exposed to high environmental temperature during the summer months. These environmental stressors may have influences on bird performance and susceptibility to pathogens such as Salmonella enteritidis by altering the intestinal micrbiota and changes in the gut integrity. Approach: This research was conducted to show that acute stressors in the poultry production can induce changes in the normal intestinal microbiota and epithelium structure and execratory functions, which may cause an increase in the opportunities of attachment of Salmonella enteritidis. Results: Experiments were conducted to determine the influence of 24 h feed withdrawal with 24 h exposure to high temperature (30°C on intestinal characteristics of broilers. Attachment of Salmonella enteritidis to ileal tissue was determined using an in vitro ileal loop assay. Changes in commensally intestinal microbial populations were determined using gel electrophoresis and alterations in ileal morphology were determined histologically. The results showed that attachment of Salmonella enteritidis to ileal tissues increased by 1.5 logs (9.05 log10 Vs 7.59 log10 Salmonella enteritidis/g of ileal tissue; p = 0.0006 in broilers fasted for 24 h also, ileal tissues from birds subjected to 30°C for 24 h had increased the attachment of Salmonella enteritidis (8.77 log10 Vs 8.50 log10 Salmonella enteritidis/g of ileum; p = 0.01 compared with birds held at 23°C. Exposure to 30°C for 24 h also altered the microbial structure in the ileum and cecum. Where subjecting birds to 30°C for 24 h reduced the crypt depth (6.0 Vs 7.8 µm, respectively; p = 0.002, but it had no effect on villus height or villus: Crypt ratio. Conclusion: The findings of the experiment explained the mechanisms by which stressors alters the

  9. Transposon mutagenesis of Salmonella enterica serovar Enteritidis identifies genes that contribute to invasiveness in human and chicken cells and survival in egg albumen.

    Science.gov (United States)

    Shah, Devendra H; Zhou, Xiaohui; Kim, Hye-Young; Call, Douglas R; Guard, Jean

    2012-12-01

    Salmonella enterica serovar Enteritidis is an important food-borne pathogen, and chickens are a primary reservoir of human infection. While most knowledge about Salmonella pathogenesis is based on research conducted on Salmonella enterica serovar Typhimurium, S. Enteritidis is known to have pathobiology specific to chickens that impacts epidemiology in humans. Therefore, more information is needed about S. Enteritidis pathobiology in comparison to that of S. Typhimurium. We used transposon mutagenesis to identify S. Enteritidis virulence genes by assay of invasiveness in human intestinal epithelial (Caco-2) cells and chicken liver (LMH) cells and survival within chicken (HD-11) macrophages as a surrogate marker for virulence. A total of 4,330 transposon insertion mutants of an invasive G1 Nal(r) strain were screened using Caco-2 cells. This led to the identification of attenuating mutations in a total of 33 different loci, many of which include genes previously known to contribute to enteric infection (e.g., Salmonella pathogenicity island 1 [SPI-1], SPI-4, SPI-5, CS54, fliH, fljB, csgB, spvR, and rfbMN) in S. Enteritidis and other Salmonella serovars. Several genes or genomic islands that have not been reported previously (e.g., SPI-14, ksgA, SEN0034, SEN2278, and SEN3503) or that are absent in S. Typhimurium or in most other Salmonella serovars (e.g., pegD, SEN1152, SEN1393, and SEN1966) were also identified. Most mutants with reduced Caco-2 cell invasiveness also showed significantly reduced invasiveness in chicken liver cells and impaired survival in chicken macrophages and in egg albumen. Consequently, these genes may play an important role during infection of the chicken host and also contribute to successful egg contamination by S. Enteritidis.

  10. Plasmid profile and pulsed-field gel electrophoresis analysis of Salmonella enterica isolates from humans in Turkey.

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    Kerem Ozdemir

    Full Text Available This study was conducted for typing Salmonella enterica subspecies enterica strains in Turkey using pulsed-field gel electrophoresis (PFGE and plasmid DNA profile analysis. Fourty-two strains were isolated from clinical samples obtained from unrelated patients with acute diarrhea. The samples were collected from state hospitals and public health laboratories located at seven provinces in different regions of Turkey at different times between 2004 and 2010. The strains were determined to belong to 4 different serovars. The Salmonella enterica strains belonged to the serovars Salmonella Enteritidis (n = 23, Salmonella Infantis (n = 14, Salmonella Munchen (n = 2, and Salmonella Typhi (n = 3. Forty-two Salmonella enterica strains were typed with PFGE methods using XbaI restriction enzyme and plasmid analysis. At the end of typing, 11 different PFGE band profiles were obtained. Four different PFGE profiles (type 1, 4, 9, and 10 were found among serotype S. Enteritidis species, 3 different PFGE profiles (type 3, 5, 6 were found among S. Infantis species, 2 different PFGE profiles were found among S. Typhi species (type 2 and 11, and 2 different PFGE profiles were found among S. Munchen species (type 7, 8. The UPGMA dendrogram was built on the PFGE profiles. In this study, it was determined that 4 strains of 42 Salmonella enterica strains possess no plasmid, while the isolates have 1-3 plasmids ranging from 5.0 to 150 kb and making 12 different plasmid profiles (P1-P12. In this study, we have applied the analysis of the PFGE patterns and used bioinformatics methods to identify both inter and intra serotype relationships of 4 frequently encountered serotypes for the first time in Turkey.

  11. Prevalence and characterization of multi-drug resistant Salmonella Enterica serovar Gallinarum biovar Pullorum and Gallinarum from chicken

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    Md. Shafiullah Parvej

    2016-01-01

    Full Text Available Aim: Salmonella is an important zoonotic pathogen responsible for animal and human diseases. The aim of the present study was to determine the prevalence and stereotyping of Salmonella isolates isolated from apparently healthy poultry. Furthermore, the clonal relatedness among the isolated Salmonella serovars was assessed. Materials and Methods: A total of 150 cloacal swab samples from apparently healthy chickens were collected, and were subjected for the isolation and identification of associated Salmonella organisms. The isolated colonies were identified and characterized on the basis of morphology, cultural characters, biochemical tests, slide agglutination test, polymerase chain reaction, and pulsed-field gel electrophoresis (PFGE. Antibiotic sensitivity patterns were also investigated using commonly used antibiotics. Results: Of the 150 samples, 11 (7.33% produced characteristics pink colony with black center on XLD agar medium, and all were culturally and biochemically confirmed to be Salmonella. All possessed serovar-specific gene SpeF and reacted uniformly with group D antisera, suggesting that all of the isolates were Salmonella Enterica serovar Gallinarum, biovar Pullorum and/or Gallinarum. Antimicrobial susceptibility testing revealed that 54.54% of the isolated Salmonella Enterica serovars were highly sensitive to ciprofloxacin, whereas the 81.81% isolates were resistant to amoxycillin, doxycycline, kanamycin, gentamycin, and tetracycline. Pulsed-field gel electrophoresis of the XbaI-digested genomic DNA exhibited identical banding patterns, suggesting that the multidrug resistant Salmonella Enterica serovars occurring in commercial layers are highly clonal in Bangladesh. Conclusion: The present study was conducted to find out the prevalence of poultry Salmonella in layer chicken and to find out the clonal relationship among them. The data in this study suggest the prevalence of Salmonella Enterica, which is multidrug resistant and

  12. A Phylogenetic and Phenotypic Analysis of Salmonella enterica Serovar Weltevreden, an Emerging Agent of Diarrheal Disease in Tropical Regions.

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    Carine Makendi

    2016-02-01

    Full Text Available Salmonella enterica serovar Weltevreden (S. Weltevreden is an emerging cause of diarrheal and invasive disease in humans residing in tropical regions. Despite the regional and international emergence of this Salmonella serovar, relatively little is known about its genetic diversity, genomics or virulence potential in model systems. Here we used whole genome sequencing and bioinformatics analyses to define the phylogenetic structure of a diverse global selection of S. Weltevreden. Phylogenetic analysis of more than 100 isolates demonstrated that the population of S. Weltevreden can be segregated into two main phylogenetic clusters, one associated predominantly with continental Southeast Asia and the other more internationally dispersed. Subcluster analysis suggested the local evolution of S. Weltevreden within specific geographical regions. Four of the isolates were sequenced using long read sequencing to produce high quality reference genomes. Phenotypic analysis in Hep-2 cells and in a murine infection model indicated that S. Weltevreden were significantly attenuated in these models compared to the classical S. Typhimurium reference strain SL1344. Our work outlines novel insights into this important emerging pathogen and provides a baseline understanding for future research studies.

  13. A Phylogenetic and Phenotypic Analysis of Salmonella enterica Serovar Weltevreden, an Emerging Agent of Diarrheal Disease in Tropical Regions.

    Science.gov (United States)

    Makendi, Carine; Page, Andrew J; Wren, Brendan W; Le Thi Phuong, Tu; Clare, Simon; Hale, Christine; Goulding, David; Klemm, Elizabeth J; Pickard, Derek; Okoro, Chinyere; Hunt, Martin; Thompson, Corinne N; Phu Huong Lan, Nguyen; Tran Do Hoang, Nhu; Thwaites, Guy E; Le Hello, Simon; Brisabois, Anne; Weill, François-Xavier; Baker, Stephen; Dougan, Gordon

    2016-02-01

    Salmonella enterica serovar Weltevreden (S. Weltevreden) is an emerging cause of diarrheal and invasive disease in humans residing in tropical regions. Despite the regional and international emergence of this Salmonella serovar, relatively little is known about its genetic diversity, genomics or virulence potential in model systems. Here we used whole genome sequencing and bioinformatics analyses to define the phylogenetic structure of a diverse global selection of S. Weltevreden. Phylogenetic analysis of more than 100 isolates demonstrated that the population of S. Weltevreden can be segregated into two main phylogenetic clusters, one associated predominantly with continental Southeast Asia and the other more internationally dispersed. Subcluster analysis suggested the local evolution of S. Weltevreden within specific geographical regions. Four of the isolates were sequenced using long read sequencing to produce high quality reference genomes. Phenotypic analysis in Hep-2 cells and in a murine infection model indicated that S. Weltevreden were significantly attenuated in these models compared to the classical S. Typhimurium reference strain SL1344. Our work outlines novel insights into this important emerging pathogen and provides a baseline understanding for future research studies.

  14. A Phylogenetic and Phenotypic Analysis of Salmonella enterica Serovar Weltevreden, an Emerging Agent of Diarrheal Disease in Tropical Regions.

    Science.gov (United States)

    Makendi, Carine; Page, Andrew J; Wren, Brendan W; Le Thi Phuong, Tu; Clare, Simon; Hale, Christine; Goulding, David; Klemm, Elizabeth J; Pickard, Derek; Okoro, Chinyere; Hunt, Martin; Thompson, Corinne N; Phu Huong Lan, Nguyen; Tran Do Hoang, Nhu; Thwaites, Guy E; Le Hello, Simon; Brisabois, Anne; Weill, François-Xavier; Baker, Stephen; Dougan, Gordon

    2016-02-01

    Salmonella enterica serovar Weltevreden (S. Weltevreden) is an emerging cause of diarrheal and invasive disease in humans residing in tropical regions. Despite the regional and international emergence of this Salmonella serovar, relatively little is known about its genetic diversity, genomics or virulence potential in model systems. Here we used whole genome sequencing and bioinformatics analyses to define the phylogenetic structure of a diverse global selection of S. Weltevreden. Phylogenetic analysis of more than 100 isolates demonstrated that the population of S. Weltevreden can be segregated into two main phylogenetic clusters, one associated predominantly with continental Southeast Asia and the other more internationally dispersed. Subcluster analysis suggested the local evolution of S. Weltevreden within specific geographical regions. Four of the isolates were sequenced using long read sequencing to produce high quality reference genomes. Phenotypic analysis in Hep-2 cells and in a murine infection model indicated that S. Weltevreden were significantly attenuated in these models compared to the classical S. Typhimurium reference strain SL1344. Our work outlines novel insights into this important emerging pathogen and provides a baseline understanding for future research studies. PMID:26867150

  15. The Vi capsular polysaccharide enables Salmonella enterica serovar typhi to evade microbe-guided neutrophil chemotaxis.

    Directory of Open Access Journals (Sweden)

    Tamding Wangdi

    2014-08-01

    Full Text Available Salmonella enterica serovar Typhi (S. Typhi causes typhoid fever, a disseminated infection, while the closely related pathogen S. enterica serovar Typhimurium (S. Typhimurium is associated with a localized gastroenteritis in humans. Here we investigated whether both pathogens differ in the chemotactic response they induce in neutrophils using a single-cell experimental approach. Surprisingly, neutrophils extended chemotactic pseudopodia toward Escherichia coli and S. Typhimurium, but not toward S. Typhi. Bacterial-guided chemotaxis was dependent on the presence of complement component 5a (C5a and C5a receptor (C5aR. Deletion of S. Typhi capsule biosynthesis genes markedly enhanced the chemotactic response of neutrophils in vitro. Furthermore, deletion of capsule biosynthesis genes heightened the association of S. Typhi with neutrophils in vivo through a C5aR-dependent mechanism. Collectively, these data suggest that expression of the virulence-associated (Vi capsular polysaccharide of S. Typhi obstructs bacterial-guided neutrophil chemotaxis.

  16. Outbreak of Salmonella enterica serovar Typhimurium phage type DT41 in Danish poultry production

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Hintzmann, Ann-Sofie; Sørensen, Gitte;

    2015-01-01

    . Typhimurium DT41 was noted mainly in this production, but also in other samples. To investigate this is in more detail, 47 isolates from egg layers (n = 5, 1 flock), broilers (n = 33, 13 flocks), broiler breeding flocks and hatches (n = 5; 2 flocks and 1 environmental hatchery sample), feed (n = 1), poultry......Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) is one of the most prevalent serovars in Europe - where both poultry and poultry related products are common sources of human salmonellosis. Due to efficient control programs, the prevalence of S. Typhimurium in Danish...... poultry production is very low. Despite this, during the past decades there has been a reoccurring problem with infections with S. Typhimurium phage type DT41 in the Danish poultry production without identifying a clear source. In the end of 2013 and beginning of 2014 an increased isolation of S...

  17. The Vi capsular polysaccharide enables Salmonella enterica serovar typhi to evade microbe-guided neutrophil chemotaxis.

    Science.gov (United States)

    Wangdi, Tamding; Lee, Cheng-Yuk; Spees, Alanna M; Yu, Chenzhou; Kingsbury, Dawn D; Winter, Sebastian E; Hastey, Christine J; Wilson, R Paul; Heinrich, Volkmar; Bäumler, Andreas J

    2014-08-01

    Salmonella enterica serovar Typhi (S. Typhi) causes typhoid fever, a disseminated infection, while the closely related pathogen S. enterica serovar Typhimurium (S. Typhimurium) is associated with a localized gastroenteritis in humans. Here we investigated whether both pathogens differ in the chemotactic response they induce in neutrophils using a single-cell experimental approach. Surprisingly, neutrophils extended chemotactic pseudopodia toward Escherichia coli and S. Typhimurium, but not toward S. Typhi. Bacterial-guided chemotaxis was dependent on the presence of complement component 5a (C5a) and C5a receptor (C5aR). Deletion of S. Typhi capsule biosynthesis genes markedly enhanced the chemotactic response of neutrophils in vitro. Furthermore, deletion of capsule biosynthesis genes heightened the association of S. Typhi with neutrophils in vivo through a C5aR-dependent mechanism. Collectively, these data suggest that expression of the virulence-associated (Vi) capsular polysaccharide of S. Typhi obstructs bacterial-guided neutrophil chemotaxis.

  18. Nation-wide Salmonella enterica surveillance and control in Danish slaughter swine herds

    DEFF Research Database (Denmark)

    Mousing, Jan; Jensen, P.T.; Halgaard, C.;

    1997-01-01

    A nation-wide Salmonella enterica surveillance and control programme was initiated in Danish finishing herds over the first quarter of 1995, In Denmark, all swine for slaughter are identifiable by a unique herd code. For each herd code, and depending on the herd's annual kill, random samples...... management hygiene precautions must be taken. If a herd is categorized in Level 3, the finishers from the herd must additionally be slaughtered under special hygiene precautions. This is supervised by the veterinary authorities. During 1995, 604 000 samples were tested for S. enterica, corresponding to 3...... during 1995 ranged from a mean of 2.9% in smaller herds (101-200 swine slaughtered per year) to 6.1% in relatively large herds (more than 5000 swine slaughtered per year)....

  19. Serotypes and Antimicrobial Resistance of Human Nontyphoidal Isolates of Salmonella enterica from Crete, Greece

    Directory of Open Access Journals (Sweden)

    Sofia Maraki

    2014-01-01

    Full Text Available We report on the serotype distribution and the antimicrobial resistance patterns to 20 different antimicrobials of 150 Salmonella enterica strains isolated from stools of diarrhoeal patients on the island of Crete over the period January 2011-December 2012. Among the S. enterica serotypes recovered, Enteritidis was the most prevalent (37.3%, followed by Typhimurium (28.7% and Newport (8.7%. No resistance was detected to extended-spectrum cephalosporins and carbapenems. Rates of resistance to ampicillin, amoxicillin/clavulanic acid, chloramphenicol, tetracycline, and cotrimoxazole were 9.3%, 4%, 2%, 15.3%, and 8.7%, respectively. Resistance to ≥4 antibiotics was primarily observed for serotypes Typhimurium and Hadar. Enteritidis remains the predominant serotype in Crete. Although low resistance to most antimicrobials was detected, continued surveillance of susceptibility is needed due to the risk of resistance.

  20. Isolation and Characterization of Antimicrobial-Resistant Nontyphoidal Salmonella enterica Serovars from Imported Food Products.

    Science.gov (United States)

    Bae, Dongryeoul; Kweon, Ohgew; Khan, Ashraf A

    2016-08-01

    The objective of this study was to determine antimicrobial resistance and elucidate the resistance mechanism in nontyphoidal Salmonella enterica serovars isolated from food products imported into the United States from 2011 to 2013. Food products contaminated with antimicrobial-resistant nontyphoidal S. enterica were mainly imported from Taiwan, Indonesia, Vietnam, and China. PCR, DNA sequencing, and plasmid analyses were used to characterize antimicrobial resistance determinants. Twentythree of 110 S. enterica isolates were resistant to various antimicrobial classes, including β-lactam, aminoglycoside, phenicol, glycopeptide, sulfonamide, trimethoprim, and/or fluoroquinolone antimicrobial agents. Twelve of the isolates were multidrug resistant strains. Antimicrobial resistance determinants blaTEM-1, blaCTX-M-9, blaOXA-1, tetA, tetB, tetD, dfrA1, dfrV, dhfrI, dhfrXII, drf17, aadA1, aadA2, aadA5, orfC, qnrS, and mutations of gyrA and parC were detected in one or more antimicrobial-resistant nontyphoidal S. enterica strains. Plasmid profiles revealed that 12 of the 23 antimicrobial-resistant strains harbored plasmids with incompatibility groups IncFIB, IncHI1, IncI1, IncN, IncW, and IncX. Epidemiologic and antimicrobial resistance monitoring data combined with molecular characterization of antimicrobial resistance determinants in Salmonella strains isolated from imported food products may provide information that can be used to establish or implement food safety programs to improve public health.

  1. Molecular Characterisation of Salmonella enterica Serovar Typhi Isolated from Typhoidial Humans

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    Arunava Das

    2012-09-01

    Full Text Available Aims: Salmonella enterica serovar Typhi is the major causative agent for typhoidial fever around the globe among human population reported till date. Present research work was carried out for detection and molecular characterisation of Salmonella enterica serovar Typhi isolated from humans with Typhoidial fever by biochemical, phenotypical and virulence gene based polymerase chain reaction (PCR techniques. The isolated strains were also investigated for antibiotic susceptibility patterns as a control measure. Methodology and Results: A total of 16 clinical samples were collected from the same numbers of patients (7 males and 9 females from Coimbatore, Erode and Salem districts of Tamil Nadu and were processed via broth enrichment methods for isolation and identification of the causative agent S. enterica serovar Typhi. Microbiological and biochemical investigations revealed the presence of S. Typhi from 16 samples. The biotyping of the isolates showed that all the isolates belonged to biotype IV. The PCR analysis confirmed the presence of invA (Invasion gene, 244bp, tyv (Tyveloseepimerase gene, 615 bp, fliC-d (Phage-1 flagellin gene for d-antigen, 750 bp and viaB (Vi antigen gene, 439bp in all 16 clinical samples. The antibiotic susceptibility test that was carried out among the isolates against 12 antimicrobial agents, showed 100 % resistance to only ampicillin and 100 % sensitivity to carbenicillin, chloramphenicol, clindamycin, gentamycin, kanamycin and tetracycline.Conclusion, significance and impact of study: This study confirmed the association of virulent strains of S. enterica serovar Typhi from Typhoidial fever among human population and suggested that PCR based diagnostic could be very useful for the rapid detection of S. Typhi isolates. Present study emphasized the use of antibiotic like chloramphenicol or in combination with other antibiotics for the effective control of S. Typhi.

  2. Isolation and Characterization of Antimicrobial-Resistant Nontyphoidal Salmonella enterica Serovars from Imported Food Products.

    Science.gov (United States)

    Bae, Dongryeoul; Kweon, Ohgew; Khan, Ashraf A

    2016-08-01

    The objective of this study was to determine antimicrobial resistance and elucidate the resistance mechanism in nontyphoidal Salmonella enterica serovars isolated from food products imported into the United States from 2011 to 2013. Food products contaminated with antimicrobial-resistant nontyphoidal S. enterica were mainly imported from Taiwan, Indonesia, Vietnam, and China. PCR, DNA sequencing, and plasmid analyses were used to characterize antimicrobial resistance determinants. Twentythree of 110 S. enterica isolates were resistant to various antimicrobial classes, including β-lactam, aminoglycoside, phenicol, glycopeptide, sulfonamide, trimethoprim, and/or fluoroquinolone antimicrobial agents. Twelve of the isolates were multidrug resistant strains. Antimicrobial resistance determinants blaTEM-1, blaCTX-M-9, blaOXA-1, tetA, tetB, tetD, dfrA1, dfrV, dhfrI, dhfrXII, drf17, aadA1, aadA2, aadA5, orfC, qnrS, and mutations of gyrA and parC were detected in one or more antimicrobial-resistant nontyphoidal S. enterica strains. Plasmid profiles revealed that 12 of the 23 antimicrobial-resistant strains harbored plasmids with incompatibility groups IncFIB, IncHI1, IncI1, IncN, IncW, and IncX. Epidemiologic and antimicrobial resistance monitoring data combined with molecular characterization of antimicrobial resistance determinants in Salmonella strains isolated from imported food products may provide information that can be used to establish or implement food safety programs to improve public health. PMID:27497122

  3. 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

    Full Text Available BACKGROUND: 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. METHODS: 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. PRINCIPAL FINDINGS: 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. CONCLUSION: We have described a simple yet effective PCR method to support surveillance of the incidence of invasive disease caused by NTS in developing countries.

  4. Receptor diversity and host interaction of bacteriophages infecting Salmonella enterica serovar Typhimurium.

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    Hakdong Shin

    Full Text Available BACKGROUND: Salmonella enterica subspecies enterica serovar Typhimurium is a gram-negative pathogen causing salmonellosis. Salmonella Typhimurium-targeting bacteriophages have been proposed as an alternative biocontrol agent to antibiotics. To further understand infection and interaction mechanisms between the host strains and the bacteriophages, the receptor diversity of these phages needs to be elucidated. METHODOLOGY/PRINCIPAL FINDINGS: Twenty-five Salmonella phages were isolated and their receptors were identified by screening a Tn5 random mutant library of S. Typhimurium SL1344. Among them, three types of receptors were identified flagella (11 phages, vitamin B(12 uptake outer membrane protein, BtuB (7 phages and lipopolysaccharide-related O-antigen (7 phages. TEM observation revealed that the phages using flagella (group F or BtuB (group B as a receptor belong to Siphoviridae family, and the phages using O-antigen of LPS as a receptor (group L belong to Podoviridae family. Interestingly, while some of group F phages (F-I target FliC host receptor, others (F-II target both FliC and FljB receptors, suggesting that two subgroups are present in group F phages. Cross-resistance assay of group B and L revealed that group L phages could not infect group B phage-resistant strains and reversely group B phages could not infect group L SPN9TCW-resistant strain. CONCLUSIONS/SIGNIFICANCE: In this report, three receptor groups of 25 newly isolated S. Typhimurium-targeting phages were determined. Among them, two subgroups of group F phages interact with their host receptors in different manner. In addition, the host receptors of group B or group L SPN9TCW phages hinder other group phage infection, probably due to interaction between receptors of their groups. This study provides novel insights into phage-host receptor interaction for Salmonella phages and will inform development of optimal phage therapy for protection against Salmonella.

  5. DNA sequence analysis of plasmids from multidrug resistant Salmonella enterica serotype Heidelberg isolates.

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    Jing Han

    Full Text Available Salmonella enterica serovar Heidelberg is among the most detected serovars in swine and poultry, ranks among the top five serotypes associated with human salmonellosis and is disproportionately associated with invasive infections and mortality in humans. Salmonella are known to carry plasmids associated with antimicrobial resistance and virulence. To identify plasmid-associated genes in multidrug resistant S. enterica serovar Heidelberg, antimicrobial resistance plasmids from five isolates were sequenced using the 454 LifeSciences pyrosequencing technology. Four of the isolates contained incompatibility group (Inc A/C multidrug resistance plasmids harboring at least eight antimicrobial resistance genes. Each of these strains also carried a second resistance plasmid including two IncFIB, an IncHI2 and a plasmid lacking an identified Inc group. The fifth isolate contained an IncI1 plasmid, encoding resistance to gentamicin, streptomycin and sulfonamides. Some of the IncA/C plasmids lacked the full concert of transfer genes and yet were able to be conjugally transferred, likely due to the transfer genes carried on the companion plasmids in the strains. Several non-IncA/C resistance plasmids also carried putative virulence genes. When the sequences were compared to previously sequenced plasmids, it was found that while all plasmids demonstrated some similarity to other plasmids, they were unique, often due to differences in mobile genetic elements in the plasmids. Our study suggests that Salmonella Heidelberg isolates harbor plasmids that co-select for antimicrobial resistance and virulence, along with genes that can mediate the transfer of plasmids within and among other bacterial isolates. Prevalence of such plasmids can complicate efforts to control the spread of S. enterica serovar Heidelberg in food animal and human populations.

  6. Competition of Lactobacillus paracasei with Salmonella enterica for Adhesion to Caco-2 Cells

    Directory of Open Access Journals (Sweden)

    Alicja Jankowska

    2008-01-01

    Full Text Available Competition of commensal and probiotic bacteria with pathogens for adhesion and colonization is one of the important protective mechanisms of gastrointestinal tract. In this study, we examined the ability of Lactobacillus paracasei to inhibit the adhesion of pathogenic Salmonella enterica to human colon adenocarcinoma Caco-2 cells. Caco-2 cells were grown for 6 or 21 days to obtain nondifferentiated or well-differentiated cells, respectively. In adhesion experiments, bacteria were added to the cells for 2 or 4 hours. The number of attached bacteria was expressed as colony-forming units (CFUs, Caco-2 cells were counted in hematocytometer. Both bacterial strains used adhered better to well-differentiated than to nondifferentiated Caco-2 cells, however, the amount of Salmonella adhered to Caco-2 after 2 hours of contact was 12-fold higher in comparison to . paracasei and almost 27-fold higher after 4 hours of contact. Two types of experiments were done: coincubation (both bacteria were added to Caco-2 cells simultaneously, and preincubation (. paracasei was incubated with Caco-2 cells first, and then . enterica was added. In coincubation experiment, the presence of . paracasei decreased . enterica adhesion by 4-fold and in preincubation experiment even 7-fold. Generally, Lactobacillus spent culture supernatants (SCSs acted weaker as inhibitors of Salmonella adhesion in comparison to the whole . paracasei culture in coincubation experiment. In conclusion, the displacement of pathogens by lactic acid bacteria and its secretions showed here depends on the time of bacteria-epithelial cell contact, and also on the stage of Caco-2 differentiation.

  7. 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.

  8. Salmonella enterica serovar Ohio septic arthritis and bone abscess in an immunocompetent patient: a case report

    Directory of Open Access Journals (Sweden)

    Kato Hideaki

    2012-07-01

    Full Text Available Abstract Introduction Non-typhi Salmonella species cause severe extra-intestinal focal infection after occult bacteremia. Although the number of cases of non-typhi salmonellosis is increasing worldwide among patients with immunocompromising conditions such as human immunodeficiency virus infection, infection is uncommon in immunocompetent subjects. We report a case of septic arthritis and bone abscess due to a rare non-typhi Salmonella organism that developed after a prolonged asymptomatic period. Case presentation A 44-year-old Japanese immunocompetent man presented with acute-onset left knee pain and swelling. He had no history of food poisoning, and his most recent travel to an endemic area was 19 years ago. Salmonella enterica serovar Ohio was identified from samples of bone abscess and joint tissue. Arthrotomy and necrotic tissue debridement followed by intravenous ceftriaxone was successful. Conclusions Non-typhi Salmonella species only rarely cause extra-intestinal focal infections in immunocompetent patients. Our case suggests that non-typhi Salmonella species can cause severe focal infections many years after the occult bacteremia associated with food poisoning.

  9. 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...

  10. Extended-Spectrum Cephalosporin-Resistant Salmonella enterica serovar Heidelberg Strains, the Netherlands1

    Science.gov (United States)

    Geurts, Yvon; Dierikx, Cindy M.; Brouwer, Michael S.M.; Kant, Arie; Wit, Ben; Heymans, Raymond; van Pelt, Wilfrid; Mevius, Dik J.

    2016-01-01

    Extended-spectrum cephalosporin-resistant Salmonella enterica serovar Heidelberg strains (JF6X01.0022/XbaI.0251, JF6X01.0326/XbaI.1966, JF6X01.0258/XbaI.1968, and JF6X01.0045/XbaI.1970) have been identified in the United States with pulsed-field gel electrophoresis. Our examination of isolates showed introduction of these strains in the Netherlands and highlight the need for active surveillance and intervention strategies by public health organizations. PMID:27314180

  11. Extended-Spectrum Cephalosporin-Resistant Salmonella enterica serovar Heidelberg Strains, the Netherlands(1).

    Science.gov (United States)

    Liakopoulos, Apostolos; Geurts, Yvon; Dierikx, Cindy M; Brouwer, Michael S M; Kant, Arie; Wit, Ben; Heymans, Raymond; van Pelt, Wilfrid; Mevius, Dik J

    2016-07-01

    Extended-spectrum cephalosporin-resistant Salmonella enterica serovar Heidelberg strains (JF6X01.0022/XbaI.0251, JF6X01.0326/XbaI.1966, JF6X01.0258/XbaI.1968, and JF6X01.0045/XbaI.1970) have been identified in the United States with pulsed-field gel electrophoresis. Our examination of isolates showed introduction of these strains in the Netherlands and highlight the need for active surveillance and intervention strategies by public health organizations. PMID:27314180

  12. Salmonella enterica Serotype Typhimurium Fimbrial Proteins Serve as Antigens during Infection of Mice

    OpenAIRE

    Humphries, Andrea; DeRidder, Sandra; Bäumler, Andreas J.

    2005-01-01

    The Salmonella enterica serotype Typhimurium genome contains 13 operons with homology to fimbrial gene sequences. Here we investigated the role of 11 serotype Typhimurium fimbrial proteins, including FimA, AgfA (CsgA), BcfA, StbA, SthA, LpfA, PefA, StdA, StcA, StiA, and StfA, as antigens during the infection of genetically resistant mice (CBA). Upon the growth of serotype Typhimurium in standard laboratory broth culture, only the expression of FimA could be detected by Western blot analysis. ...

  13. First isolation of Salmonella enterica serovar Napoli from wild birds in Italy

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    Laura Mancini

    2014-03-01

    Full Text Available Salmonella enterica serovar Napoli (S. Napoli is an emerging serovar in Italy. It accounts for 2-4% of all serovars isolated from human infections. The zoonotic origin of this serovar is still unknown and this makes difficult to apply any control intervention. We report here the isolation of S. Napoli from a river nightingale (Cettia cetti, Temminck 1820 which represents the first description of this serovar from wild birds. This finding adds knowledge to the ecology of S. Napoli and addresses further studies aimed to assess the epidemiologic link between S. Napoli isolated from wild birds, food, environmental sources and human infections.

  14. Evidence that a B12-Adenosyl Transferase Is Encoded within the Ethanolamine Operon of Salmonella enterica

    OpenAIRE

    Sheppard, David E.; Penrod, Joseph T.; Bobik, Thomas; Kofoid, Eric; Roth, John R.

    2004-01-01

    Adenosylcobalamin (Ado-B12) is both the cofactor and inducer of ethanolamine ammonia lyase (EA-lyase), a catabolic enzyme for ethanolamine. De novo synthesis of Ado-B12 by Salmonella enterica occurs only under anaerobic conditions. Therefore, aerobic growth on ethanolamine requires import of Ado-B12 or a precursor (CN-B12 or OH-B12) that can be adenosylated internally. Several known enzymes adenosylate corrinoids. The CobA enzyme transfers adenosine from ATP to a biosynthetic intermediate in ...

  15. An Improved Method to Knock Out the asd Gene of Salmonella enterica Serovar Pullorum

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    Shi-Zhong Geng

    2009-01-01

    Full Text Available An asd-deleted (Δasd mutant of Salmonella enterica serovar Pullorum (SP was constructed using an improved method of gene knockout by combining the π-suicide plasmid system with the Red Disruption system. The asd gene was efficiently knocked out by the recombinant suicide vector, which replaced the asd gene with the CmR gene. Based on the balanced lethal host-vector system, the phenotype of the Δasd mutant was further defined. The improved method was simpler and more effective than previously reported conventional methods.

  16. National surveillance of Salmonella enterica in food-producing animals in Japan

    OpenAIRE

    Kijima Mayumi; Kojima Akemi; Morioka Ayako; Takahashi Toshio; Ishihara Kanako; Asai Tetsuo; Tamura Yutaka

    2009-01-01

    Abstract A total of 518 fecal samples collected from 183 apparently healthy cattle, 180 pigs and 155 broilers throughout Japan in 1999 were examined to determine the prevalence and antimicrobial susceptibility of Salmonella. The isolation rates were 36.1% in broilers, 2.8% in pigs and 0.5% in cattle. S. enterica Infantis was the most frequent isolate, found in 22.6% of broiler fecal samples. Higher resistance rates were observed against oxytetracycline (82.0%), dihydrostreptomycin (77.9%), ka...

  17. Differential induction of total IgE by two Salmonella enterica serotypes

    DEFF Research Database (Denmark)

    Ktsoyan, Zhanna A.; Mkrtchyan, Mkhitar S.; Zakharyan, Magdalina K.;

    2015-01-01

    The main goal of this study was to establish how the inflammation caused by infection with two different Salmonella enterica serotypes, S. Typhimurium and S. Enteritidis, may lead to the predisposition to allergy as measured by total IgE level in the blood. Infection by S. Typhimurium did...... with the comparative counts of cultivated bacteria showed an inverse relationship with the IgE concentration. Earlier we reported the elevated level of IL-17 in patients infected by S. Enteritidis. In the current study a significant correlation was found between the concentrations of IL-17 and IgE suggesting...

  18. Small Molecule Restores Itaconate Sensitivity in Salmonella enterica: A Potential New Approach to Treating Bacterial Infections.

    Science.gov (United States)

    Hammerer, Fabien; Chang, Justin H; Duncan, Dustin; Castañeda Ruiz, Angel; Auclair, Karine

    2016-08-17

    In the context of increasing global antibiotic resistance, the need for alternative therapeutic targets is great. Although new antibiotics and resistance inhibitors provide temporary solutions, they are bound to become obsolete. In this work, we propose a new approach, coined "bacterio-modulation" that aims to restore macrophage potency towards bacterial strains that are able to survive in phagolysosomes. One key defense in the macrophage's arsenal is itaconate, an endogenous molecule with antimicrobial activity. Some intracellular pathogens have evolved to produce itaconate-degrading enzymes, which are required for intracellular proliferation and to promote pathogenicity. We herein present the first molecule able to resensitize Salmonella enterica to itaconate. PMID:27254798

  19. Global Genomic Epidemiology of Salmonella enterica Serovar Typhimurium DT104.

    Science.gov (United States)

    Leekitcharoenphon, Pimlapas; Hendriksen, Rene S; Le Hello, Simon; Weill, François-Xavier; Baggesen, Dorte Lau; Jun, Se-Ran; Ussery, David W; Lund, Ole; Crook, Derrick W; Wilson, Daniel J; Aarestrup, Frank M

    2016-04-01

    It has been 30 years since the initial emergence and subsequent rapid global spread of multidrug-resistant Salmonella entericaserovar Typhimurium DT104 (MDR DT104). Nonetheless, its origin and transmission route have never been revealed. We used whole-genome sequencing (WGS) and temporally structured sequence analysis within a Bayesian framework to reconstruct temporal and spatial phylogenetic trees and estimate the rates of mutation and divergence times of 315S Typhimurium DT104 isolates sampled from 1969 to 2012 from 21 countries on six continents. DT104 was estimated to have emerged initially as antimicrobial susceptible in ∼1948 (95% credible interval [CI], 1934 to 1962) and later became MDR DT104 in ∼1972 (95% CI, 1972 to 1988) through horizontal transfer of the 13-kb Salmonella genomic island 1 (SGI1) MDR region into susceptible strains already containing SGI1. This was followed by multiple transmission events, initially from central Europe and later between several European countries. An independent transmission to the United States and another to Japan occurred, and from there MDR DT104 was probably transmitted to Taiwan and Canada. An independent acquisition of resistance genes took place in Thailand in ∼1975 (95% CI, 1975 to 1990). In Denmark, WGS analysis provided evidence for transmission of the organism between herds of animals. Interestingly, the demographic history of Danish MDR DT104 provided evidence for the success of the program to eradicate Salmonellafrom pig herds in Denmark from 1996 to 2000. The results from this study refute several hypotheses on the evolution of DT104 and suggest that WGS may be useful in monitoring emerging clones and devising strategies for prevention of Salmonella infections.

  20. Global Genomic Epidemiology of Salmonella enterica Serovar Typhimurium DT104.

    Science.gov (United States)

    Leekitcharoenphon, Pimlapas; Hendriksen, Rene S; Le Hello, Simon; Weill, François-Xavier; Baggesen, Dorte Lau; Jun, Se-Ran; Ussery, David W; Lund, Ole; Crook, Derrick W; Wilson, Daniel J; Aarestrup, Frank M

    2016-04-01

    It has been 30 years since the initial emergence and subsequent rapid global spread of multidrug-resistant Salmonella entericaserovar Typhimurium DT104 (MDR DT104). Nonetheless, its origin and transmission route have never been revealed. We used whole-genome sequencing (WGS) and temporally structured sequence analysis within a Bayesian framework to reconstruct temporal and spatial phylogenetic trees and estimate the rates of mutation and divergence times of 315S Typhimurium DT104 isolates sampled from 1969 to 2012 from 21 countries on six continents. DT104 was estimated to have emerged initially as antimicrobial susceptible in ∼1948 (95% credible interval [CI], 1934 to 1962) and later became MDR DT104 in ∼1972 (95% CI, 1972 to 1988) through horizontal transfer of the 13-kb Salmonella genomic island 1 (SGI1) MDR region into susceptible strains already containing SGI1. This was followed by multiple transmission events, initially from central Europe and later between several European countries. An independent transmission to the United States and another to Japan occurred, and from there MDR DT104 was probably transmitted to Taiwan and Canada. An independent acquisition of resistance genes took place in Thailand in ∼1975 (95% CI, 1975 to 1990). In Denmark, WGS analysis provided evidence for transmission of the organism between herds of animals. Interestingly, the demographic history of Danish MDR DT104 provided evidence for the success of the program to eradicate Salmonellafrom pig herds in Denmark from 1996 to 2000. The results from this study refute several hypotheses on the evolution of DT104 and suggest that WGS may be useful in monitoring emerging clones and devising strategies for prevention of Salmonella infections. PMID:26944846

  1. Recovery of Salmonella enterica from seropositive finishing pig herds

    DEFF Research Database (Denmark)

    Wong, Danilo Lo Fo; Dahl, J.; van der Wolf, P.J.;

    2003-01-01

    the proportion of culture positive- and seropositive samples in a herd at cut-off OD% > 10 and of 0.58 at cut-off OD% > 40. Serology is a measure of historical exposure, which may or may not correlate closely to the microbiological burden at the time of sampling. Due to the low sensitivity of culture methods......, apparent 'false-positive' serological results may well represent real infections not detected by bacteriological testing. For screening purposes, serological testing provides an indication of exposure to Salmonella, which forms the basis for targeted sampling, intervention and logistic slaughter procedures....

  2. Roles of the outer membrane protein AsmA of Salmonella enterica in the control of marRAB expression and invasion of epithelial cells.

    Science.gov (United States)

    Prieto, Ana I; Hernández, Sara B; Cota, Ignacio; Pucciarelli, M Graciela; Orlov, Yuri; Ramos-Morales, Francisco; García-del Portillo, Francisco; Casadesús, Josep

    2009-06-01

    A genetic screen for suppressors of bile sensitivity in DNA adenine methylase (dam) mutants of Salmonella enterica serovar Typhimurium yielded insertions in an uncharacterized locus homologous to the Escherichia coli asmA gene. Disruption of asmA suppressed bile sensitivity also in phoP and wec mutants of S. enterica and increased the MIC of sodium deoxycholate for the parental strain ATCC 14028. Increased levels of marA mRNA were found in asmA, asmA dam, asmA phoP, and asmA wec strains of S. enterica, suggesting that lack of AsmA activates expression of the marRAB operon. Hence, asmA mutations may enhance bile resistance by inducing gene expression changes in the marRAB-controlled Mar regulon. In silico analysis of AsmA structure predicted the existence of one transmembrane domain. Biochemical analysis of subcellular fractions revealed that the asmA gene of S. enterica encodes a protein of approximately 70 kDa located in the outer membrane. Because AsmA is unrelated to known transport and/or efflux systems, we propose that activation of marRAB in asmA mutants may be a consequence of envelope reorganization. Competitive infection of BALB/c mice with asmA(+) and asmA isogenic strains indicated that lack of AsmA attenuates Salmonella virulence by the oral route but not by the intraperitoneal route. Furthermore, asmA mutants showed a reduced ability to invade epithelial cells in vitro.

  3. Discovery of novel secreted virulence factors from Salmonella enterica serovar Typhimurium by proteomic analysis of culture supernatants.

    Science.gov (United States)

    Niemann, George S; Brown, Roslyn N; Gustin, Jean K; Stufkens, Afke; Shaikh-Kidwai, Afshan S; Li, Jie; McDermott, Jason E; Brewer, Heather M; Schepmoes, Athena; Smith, Richard D; Adkins, Joshua N; Heffron, Fred

    2011-01-01

    Salmonella enterica serovar Typhimurium is a leading cause of acute gastroenteritis throughout the world. This pathogen has two type III secretion systems (TTSS) encoded in Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2) that deliver virulence factors (effectors) to the host cell cytoplasm and are required for virulence. While many effectors have been identified and at least partially characterized, the full repertoire of effectors has not been catalogued. In this proteomic study, we identified effector proteins secreted into defined minimal medium designed to induce expression of the SPI-2 TTSS and its effectors. We compared the secretomes of the parent strain to those of strains missing essential (ssaK::cat) or regulatory (ΔssaL) components of the SPI-2 TTSS. We identified 20 known SPI-2 effectors. Excluding the translocon components SseBCD, all SPI-2 effectors were biased for identification in the ΔssaL mutant, substantiating the regulatory role of SsaL in TTS. To identify novel effector proteins, we coupled our secretome data with a machine learning algorithm (SIEVE, SVM-based identification and evaluation of virulence effectors) and selected 12 candidate proteins for further characterization. Using CyaA' reporter fusions, we identified six novel type III effectors and two additional proteins that were secreted into J774 macrophages independently of a TTSS. To assess their roles in virulence, we constructed nonpolar deletions and performed a competitive index analysis from intraperitoneally infected 129/SvJ mice. Six mutants were significantly attenuated for spleen colonization. Our results also suggest that non-type III secretion mechanisms are required for full Salmonella virulence.

  4. An outbreak of gastroenteritis caused by Salmonella enterica serotype Enteritidis traced to cream cakes

    Directory of Open Access Journals (Sweden)

    Kee Tai Goh

    2011-03-01

    Full Text Available Introduction: This paper describes the epidemiological, microbiological and environmental investigations conducted during an outbreak of Salmonella gastroenteritis in Singapore.Methods: A case-control study was undertaken to identify the vehicle of transmission. Microbiological testing was performed on faecal, food and environmental samples. Isolates of Salmonella were further characterized by phage typing and ribotyping.Results: There were 216 gastroenteritis cases reported from 20 November to 4 December 2007. The causative agent was identified as Salmonella enterica subspecies enterica serotype Enteritidis for 14 out of 20 cases tested. The vehicle of transmission was traced to cream cakes produced by a bakery and sold at its retail outlets (P < 0.001, OR = 143.00, 95% Cl = 27.23–759.10. More than two-thirds of the 40 Salmonella strains isolated from hospitalized cases, food samples and asymptomatic food handlers were of phage type 1; the others reacted but did not conform to any phage type. The phage types correlated well with their unique antibiograms. The ribotype patterns of 22 selected isolates tested were highly similar, indicating genetic relatedness. The dendrogram of the strains from the outbreak showed distinct clustering and correlation compared to the non-outbreak strains, confirming a common source of infection.Discussion: The cream cakes were likely contaminated by one of the ingredients used in the icing. Cross-contamination down the production line and subsequent storage of cakes at ambient temperatures for a prolonged period before consumption could have resulted in the outbreak.

  5. Isolation and characterization of bacteriophages of Salmonella enterica serovar Pullorum.

    Science.gov (United States)

    Bao, H; Zhang, H; Wang, R

    2011-10-01

    In this study, 2 bacteriophages of Salmonella Pullorum were isolated using an enrichment protocol and the double agar layer method. They were named PSPu-95 and PSPu-4-116, respectively, against clinical isolates of Salmonella Pullorum SPu-95 and SPu-116. The host ranges of the 2 bacteriophages were determined by performing spot tests with 20 bacteria strains. Both bacteriophages had wide host ranges. Bacteriophage PSPu-95 had a lytic effect on 17 of the 20 isolates (85%), and PSPu-4-116 produced a lytic effect on 14 isolates (70%) and was the only bacteriophage that produced a clear plaque on enterotoxigenic Escherichia coli K88. Transmission electron microscopy revealed the bacteriophages belonged to the order Caudovirales. Bacteriophage PSPu-95 was a member of the family Siphoviridae, but bacteriophage PSPu-4-116 belonged to the family Myoviridae. Both had a double-stranded DNA, which was digested with HindIII or EcoRI, that was estimated to be 58.3 kbp (PSPu-95) and 45.2 kbp (PSPu-4-116) by 1% agar electrophoresis. One-step growth kinetics showed that the latent periods were all less than 20 min, and the burst size was 77.5 pfu/cell for PSPu-95 and 86 pfu/cell for PSPu-4-116. The bacteriophages were able to survive in a pH range between 4 and 10, and they were able to survive in a treatment of 70°C for 60 min. The characterizations of these 2 bacteriophages were helpful in establishing a basis for adopting the most effective bacteriophage to control bacteria in the poultry industry.

  6. Incidence and growth of Salmonella enterica on the peel and pulp of avocado (Persea americana) and custard apple (Annona squamosa).

    Science.gov (United States)

    Rezende, Ana Carolina B; Crucello, Juliana; Moreira, Rafael C; Silva, Beatriz S; Sant'Ana, Anderson S

    2016-10-17

    The aim of this study was to assess the incidence and to estimate the growth kinetic parameters (maximum growth rate, μ; lag time, λ; and maximum population, κ) of Salmonella on the peel and pulp of avocado (Perseaamericana var. americana) and custard apple (Annona squamosa L.) as affected by temperature (10-30°C). The incidence of Salmonella was assessed on the peel and pulp of the fruits (n=200 of each fruit), separately, totalizing 800 analyses. Only three samples of custard apple pulp were positive for Salmonella enterica and the three isolates recovered belonged to serotype S. Typhimurium. Salmonella was not recovered from avocado and custard apple peels and from avocado pulp. Generally, the substrate (pulp or peel) of growth did not affect μ values of S. enterica (p>0.05). Very similar μ values were found for S. enterica inoculated in custard apple and avocado. S. enterica presented the highest λ in the peel of the fruits. The growth of S. enterica resulted in larger λ in custard apple in comparison to avocado. For example, the λ of S. enterica in the pulp of custard apple and avocado were 47.0±0.78h and 10.0±3.78h, respectively. The lowest values of κ were obtained at the lower storage temperature conditions (10°C). For instance, κ values of 3.7±0.06log CFU/g and 2.9±0.03log CFU/g were obtained from the growth of S. enterica in avocado and custard apple pulps at 10°C (pavocado led to a κ value of 6.5±0.25log CFU/g, while in the peel led to a κ value of 4.6±0.23log CFU/g (pavocado comprises a better substrate than custard apple for the growth of S. enterica. The square root model fitted to the data obtained in this study and to the growth data available in the literature for other tropical low acid fruits indicated high variability in μ and λ of Salmonella. The results obtained in this study show that whole low acid tropical fruits can harbor Salmonella, and that this foodborne pathogen can not only survive but also grow both on the

  7. The effect of nitric oxide combined with fluoroquinolones against Salmonella enterica serovar Typhimurium in vitro

    Directory of Open Access Journals (Sweden)

    Coban AY

    2003-01-01

    Full Text Available Two regulons, soxRS and marRAB, are associated with resistance to quinolones or multiple antibiotic in Salmonella enterica serovar Typhimurium. These regulons are activated by nitric oxide and redox-cycling drugs, such as paraquat and cause on activation of the acrAB-encoded efflux pump. In this study, we investigated the effect of nitric oxide (NO alone and in combination with ofloxacin, ciprofloxacin, and pefloxacin against S. typhimurium clinical isolates and mutant strains in vitro. We did not observe synergistic effect against clinical isolates and SH5014 (parent strain of acr mutant, while we found synergistic effect against PP120 (soxRS mutant and SH7616 (an acr mutant S. typhimurium for all quinolones. Our results suggest that the efficiencies of some antibiotics, including ofloxacin, ciprofloxacin, and pefloxacin are decreased via activation of soxRS and marRAB regulons by NO in S. enterica serovar Typhimurium. Further studies are warranted to establish the interaction of NO with the genes of Salmonella and, with multiple antibiotic resistance.

  8. Single passage in mouse organs enhances the survival and spread of Salmonella enterica.

    Science.gov (United States)

    Dybowski, Richard; Restif, Olivier; Goupy, Alexandre; Maskell, Duncan J; Mastroeni, Piero; Grant, Andrew J

    2015-12-01

    Intravenous inoculation of Salmonella enterica serovar Typhimurium into mice is a prime experimental model of invasive salmonellosis. The use of wild-type isogenic tagged strains (WITS) in this system has revealed that bacteria undergo independent bottlenecks in the liver and spleen before establishing a systemic infection. We recently showed that those bacteria that survived the bottleneck exhibited enhanced growth when transferred to naive mice. In this study, we set out to disentangle the components of this in vivo adaptation by inoculating mice with WITS grown either in vitro or in vivo. We developed an original method to estimate the replication and killing rates of bacteria from experimental data, which involved solving the probability-generating function of a non-homogeneous birth-death-immigration process. This revealed a low initial mortality in bacteria obtained from a donor animal. Next, an analysis of WITS distributions in the livers and spleens of recipient animals indicated that in vivo-passaged bacteria started spreading between organs earlier than in vitro-grown bacteria. These results further our understanding of the influence of passage in a host on the fitness and virulence of Salmonella enterica and represent an advance in the power of investigation on the patterns and mechanisms of host-pathogen interactions.

  9. Isolation and plasmid characterization of carbapenemase (IMP-4) producing Salmonella enterica Typhimurium from cats

    Science.gov (United States)

    Abraham, Sam; O’Dea, Mark; Trott, Darren J.; Abraham, Rebecca J.; Hughes, David; Pang, Stanley; McKew, Genevieve; Cheong, Elaine Y. L.; Merlino, John; Saputra, Sugiyono; Malik, Richard; Gottlieb, Thomas

    2016-01-01

    Carbapenem-resistant Enterobacteriaceae (CRE) are a pressing public health issue due to limited therapeutic options to treat such infections. CREs have been predominantly isolated from humans and environmental samples and they are rarely reported among companion animals. In this study we report on the isolation and plasmid characterization of carbapenemase (IMP-4) producing Salmonella enterica Typhimurium from a companion animal. Carbapenemase-producing S. enterica Typhimurium carrying blaIMP-4 was identified from a systemically unwell (index) cat and three additional cats at an animal shelter. All isolates were identical and belonged to ST19. Genome sequencing revealed the acquisition of a multidrug-resistant IncHI2 plasmid (pIMP4-SEM1) that encoded resistance to nine antimicrobial classes including carbapenems and carried the blaIMP-4-qacG-aacA4-catB3 cassette array. The plasmid also encoded resistance to arsenic (MIC-150 mM). Comparative analysis revealed that the plasmid pIMP4-SEM1 showed greatest similarity to two blaIMP-8 carrying IncHI2 plasmids from Enterobacter spp. isolated from humans in China. This is the first report of CRE carrying a blaIMP-4 gene causing a clinical infection in a companion animal, with presumed nosocomial spread. This study illustrates the broader community risk entailed in escalating CRE transmission within a zoonotic species such as Salmonella, and in a cycle that encompasses humans, animals and the environment. PMID:27767038

  10. 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.

  11. Characterization of Isolates of Salmonella enterica Serovar Stanley, a Serovar Endemic to Asia and Associated with Travel

    DEFF Research Database (Denmark)

    Hendriksen, Rene S.; Le Hello, Simon; Bortolaia, Valeria;

    2012-01-01

    Salmonella enterica serovar Stanley (S. Stanley) is a common serovar in Southeast Asia and was the second most common serovar implicated in human salmonellosis in Thailand in the years 2002 to 2007. In contrast, this serovar is relatively uncommon in Europe. The objective of this study was to cha...

  12. Immersion in antimicrobial solutions reduces Salmonella enterica and Shiga toxin-producing Escherichia coli on beef cheek meat

    Science.gov (United States)

    The objective of this study was to determine the effect of immersing beef cheek meat in antimicrobial solutions on the reduction of O157:H7 Shiga toxin–producing Escherichia coli (STEC), non-O157:H7 STEC, and Salmonella enterica. Beef cheek meat was inoculated with O157:H7 STEC, non-O157:H7 STEC, an...

  13. 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

    2007-01-01

    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 s

  14. Inactivation of Salmonella enterica serovar Typhimurium and quality maintenance of cherry tomatoes treated with gaseous essential oils

    Science.gov (United States)

    The antimicrobial activity of the essential oils (EOs) from cinnamon bark, oregano, mustard and of their major components cinnamaldehyde, carvacrol, and allyl isothiocyanate (AIT) were evaluated as a gaseous treatment to reduce Salmonella enterica serovar Typhimurium in vitro and on tomatoes. In dif...

  15. Antimicrobial activity of apple, hibiscus, olive, and hydrogen peroxide formulations against Salmonella enterica on organic leafy greens

    Science.gov (United States)

    Salmonella enterica is one of the most common bacterial pathogens involved in foodborne outbreaks with fresh produce in the last decade. In an effort to discover safe antimicrobials for use on fresh produce, the objective of the present study was to evaluate the effectiveness of different antimicro...

  16. Draft Genome Sequences of 40 Salmonella enterica Serovar Typhimurium Strains Isolated from Humans and Food in Brazil

    Science.gov (United States)

    Almeida, Fernanda; Medeiros, Marta Inês Cazentini; Rodrigues, Dália Prazeres; Payne, Justin; Timme, Ruth E.

    2016-01-01

    Salmonellosis is an important health problem worldwide and Salmonella enterica serovar Typhimurium is one of the most common isolated serovars. Here, we reported the draft genomes of 40 S. Typhimurium strains isolated from humans and food in Brazil. These draft genomes will improve phylogenetic analysis and will help enhance our understanding of strains of this serovar isolated in Brazil. PMID:27660768

  17. Draft Genome Sequences of 40 Salmonella enterica Serovar Typhimurium Strains Isolated from Humans and Food in Brazil.

    Science.gov (United States)

    Almeida, Fernanda; Medeiros, Marta Inês Cazentini; Rodrigues, Dália Prazeres; Payne, Justin; Timme, Ruth E; Allard, Marc W; Falcão, Juliana Pfrimer

    2016-01-01

    Salmonellosis is an important health problem worldwide and Salmonella enterica serovar Typhimurium is one of the most common isolated serovars. Here, we reported the draft genomes of 40 S Typhimurium strains isolated from humans and food in Brazil. These draft genomes will improve phylogenetic analysis and will help enhance our understanding of strains of this serovar isolated in Brazil. PMID:27660768

  18. Inactivation of Salmonella enterica and Listeria monocytogenes in cantaloupe puree by high hydrostatic pressure with/without added ascorbic acid

    Science.gov (United States)

    The objective of this research was to evaluate and develop a method for inactivation of Salmonella enterica and Listeria monocytogenes in cantaloupe puree (CP) by high hydrostatic pressure (HHP). Cantaloupe being the most netted varieties of melons presents a greater risk of pathogen transmission. ...

  19. Multilocus sequence typing as a replacement for serotyping in Salmonella enterica.

    Directory of Open Access Journals (Sweden)

    Mark Achtman

    Full Text Available Salmonella enterica subspecies enterica is traditionally subdivided into serovars by serological and nutritional characteristics. We used Multilocus Sequence Typing (MLST to assign 4,257 isolates from 554 serovars to 1092 sequence types (STs. The majority of the isolates and many STs were grouped into 138 genetically closely related clusters called eBurstGroups (eBGs. Many eBGs correspond to a serovar, for example most Typhimurium are in eBG1 and most Enteritidis are in eBG4, but many eBGs contained more than one serovar. Furthermore, most serovars were polyphyletic and are distributed across multiple unrelated eBGs. Thus, serovar designations confounded genetically unrelated isolates and failed to recognize natural evolutionary groupings. An inability of serotyping to correctly group isolates was most apparent for Paratyphi B and its variant Java. Most Paratyphi B were included within a sub-cluster of STs belonging to eBG5, which also encompasses a separate sub-cluster of Java STs. However, diphasic Java variants were also found in two other eBGs and monophasic Java variants were in four other eBGs or STs, one of which is in subspecies salamae and a second of which includes isolates assigned to Enteritidis, Dublin and monophasic Paratyphi B. Similarly, Choleraesuis was found in eBG6 and is closely related to Paratyphi C, which is in eBG20. However, Choleraesuis var. Decatur consists of isolates from seven other, unrelated eBGs or STs. The serological assignment of these Decatur isolates to Choleraesuis likely reflects lateral gene transfer of flagellar genes between unrelated bacteria plus purifying selection. By confounding multiple evolutionary groups, serotyping can be misleading about the disease potential of S. enterica. Unlike serotyping, MLST recognizes evolutionary groupings and we recommend that Salmonella classification by serotyping should be replaced by MLST or its equivalents.

  20. Efficacy of European starling control to reduce Salmonella enterica contamination in a concentrated animal feeding operation in the Texas panhandle

    Directory of Open Access Journals (Sweden)

    Bodenchuk Michael J

    2011-02-01

    Full Text Available Abstract Background European starlings (Sturnus vulgaris are an invasive bird species known to cause damage to plant and animal agriculture. New evidence suggests starlings may also contribute to the maintenance and spread of diseases within livestock facilities. Identifying and mitigating the risk pathways that contribute to disease in livestock is necessary to reduce production losses and contamination of human food products. To better understand the impact starlings have on disease transmission to cattle we assessed the efficacy of starling control as a tool to reduce Salmonella enterica within a concentrated animal feeding operation. We matched a large facility, slated for operational control using DRC-1339 (3-chloro-4-methylaniline hydrochloride, also 3-chloro p-toluidine hydrochloride, 3-chloro-4-methylaniline, with a comparable reference facility that was not controlling birds. In both facilities, we sampled cattle feed, cattle water and cattle feces for S. enterica before and after starling control operations. Results Within the starling-controlled CAFO, detections of S. enterica contamination disappeared from feed bunks and substantially declined within water troughs following starling control operations. Within the reference facility, detections of S. enterica contamination increased substantially within feed bunks and water troughs. Starling control was not observed to reduce prevalence of S. enterica in the cattle herd. Following starling control operations, herd prevalence of S. enterica increased on the reference facility but herd prevalence of S. enterica on the starling-controlled CAFO stayed at pretreatment levels. Conclusions Within the starling-controlled facility detections of S. enterica disappeared from feed bunks and substantially declined within water troughs following control operations. Since cattle feed and water are obvious routes for the ingestion of S. enterica, starling control shows promise as a tool to help

  1. A novel secretion pathway of Salmonella enterica acts as an antivirulence modulator during salmonellosis.

    Directory of Open Access Journals (Sweden)

    Ohad Gal-Mor

    2008-04-01

    Full Text Available Salmonella spp. are Gram-negative enteropathogenic bacteria that infect a variety of vertebrate hosts. Like any other living organism, protein secretion is a fundamental process essential for various aspects of Salmonella biology. Herein we report the identification and characterization of a horizontally acquired, autonomous and previously unreported secretion pathway. In Salmonella enterica serovar Typhimurium, this novel secretion pathway is encoded by STM1669 and STM1668, designated zirT and zirS, respectively. We show that ZirT is localized to the bacterial outer membrane, expected to adopt a compact beta-barrel conformation, and functions as a translocator for ZirS. ZirS is an exoprotein, which is secreted into the extracellular environment in a ZirT-dependent manner. The ZirTS secretion pathway was found to share several important features with two-partner secretion (TPS systems and members of the intimin/invasin family of adhesions. We show that zirTS expression is affected by zinc; and that in vivo, induction of zirT occurs distinctively in Salmonella colonizing the small intestine, but not in systemic sites. Additionally, strong expression of zirT takes place in Salmonella shed in fecal pellets during acute and persistent infections of mice. Inactivation of ZirTS results in a hypervirulence phenotype of Salmonella during oral infection of mice. Cumulatively, these results indicate that the ZirTS pathway plays a unique role as an antivirulence modulator during systemic disease and is involved in fine-tuning a host-pathogen balance during salmonellosis.

  2. Cadmium resistance and uptake by bacterium, Salmonella enterica 43C, isolated from industrial effluent.

    Science.gov (United States)

    Khan, Zaman; Rehman, Abdul; Hussain, Syed Z; Nisar, Muhammad A; Zulfiqar, Soumble; Shakoori, Abdul R

    2016-12-01

    Cadmium resistant bacterium, isolated from industrial wastewater, was characterized as Salmonella enterica 43C on the basis of biochemical and 16S rRNA ribotyping. It is first ever reported S. enterica 43C bared extreme resistance against heavy metal consortia in order of Pb(2+)>Cd(2+)>As(3+)>Zn(2+)>Cr(6+)>Cu(2+)>Hg(2+). Cd(2+) stress altered growth pattern of the bacterium in time dependent manner. It could remove nearly 57 % Cd(2+) from the medium over a period of 8 days. Kinetic and thermodynamic studies based on various adsorption isotherm models (Langmuir and Freundlich) depicted the Cd(2+) biosorption as spontaneous, feasible and endothermic in nature. Interestingly, the bacterium followed pseudo first order kinetics, making it a good biosorbent for heavy metal ions. The S. enterica 43C Cd(2+) processivity was significantly influenced by temperature, pH, initial Cd(2+) concentration, biomass dosage and co-metal ions. FTIR analysis of the bacterium revealed the active participation of amide and carbonyl moieties in Cd(2+) adsorption confirmed by EDX analysis. Electron micrographs beckoned further surface adsorption and increased bacterial size due to intracellular Cd(2+) accumulation. An overwhelming increase in glutathione and other non-protein thiols levels played a significant role in thriving oxidative stress generated by metal cations. Presence of metallothionein clearly depicted the role of such proteins in bacterial metal resistance mechanism. The present study results clearly declare S. enterica 43C a suitable candidate for green chemistry to bioremediate environmental Cd(2+).

  3. Cadmium resistance and uptake by bacterium, Salmonella enterica 43C, isolated from industrial effluent.

    Science.gov (United States)

    Khan, Zaman; Rehman, Abdul; Hussain, Syed Z; Nisar, Muhammad A; Zulfiqar, Soumble; Shakoori, Abdul R

    2016-12-01

    Cadmium resistant bacterium, isolated from industrial wastewater, was characterized as Salmonella enterica 43C on the basis of biochemical and 16S rRNA ribotyping. It is first ever reported S. enterica 43C bared extreme resistance against heavy metal consortia in order of Pb(2+)>Cd(2+)>As(3+)>Zn(2+)>Cr(6+)>Cu(2+)>Hg(2+). Cd(2+) stress altered growth pattern of the bacterium in time dependent manner. It could remove nearly 57 % Cd(2+) from the medium over a period of 8 days. Kinetic and thermodynamic studies based on various adsorption isotherm models (Langmuir and Freundlich) depicted the Cd(2+) biosorption as spontaneous, feasible and endothermic in nature. Interestingly, the bacterium followed pseudo first order kinetics, making it a good biosorbent for heavy metal ions. The S. enterica 43C Cd(2+) processivity was significantly influenced by temperature, pH, initial Cd(2+) concentration, biomass dosage and co-metal ions. FTIR analysis of the bacterium revealed the active participation of amide and carbonyl moieties in Cd(2+) adsorption confirmed by EDX analysis. Electron micrographs beckoned further surface adsorption and increased bacterial size due to intracellular Cd(2+) accumulation. An overwhelming increase in glutathione and other non-protein thiols levels played a significant role in thriving oxidative stress generated by metal cations. Presence of metallothionein clearly depicted the role of such proteins in bacterial metal resistance mechanism. The present study results clearly declare S. enterica 43C a suitable candidate for green chemistry to bioremediate environmental Cd(2+). PMID:27491862

  4. Tandem attenuators control expression of the Salmonella mgtCBR virulence operon.

    Science.gov (United States)

    Lee, Eun-Jin; Groisman, Eduardo A

    2012-10-01

    The mgtCBR operon from Salmonella enterica serovar Typhimurium specifies the virulence protein MgtC, the Mg(2+) transporter MgtB and the regulatory peptide MgtR. The mgtCBR transcript includes a long leader region harbouring two short open reading frames (ORFs). Translation of these ORFs is anticipated to impact the formation of particular stem-loop structures and control transcription of the coding region by an attenuation-like mechanism. We previously reported that ORF mgtM enables Salmonella to promote transcription of the mgtC and mgtB coding regions when experiencing a rise in cytoplasmic ATP levels. We now show that the proline codon-rich ORF mgtP mediates an increase in transcription of the mgtC and mgtB coding regions under conditions predicted to decrease the levels of proline-charged tRNA(Pro) . The high ATP and low proline signals act independently in an additive form. Replacing conserved mgtP proline codons with codons specifying other amino acids abolished the response to proline limitation but had no effect on the response to ATP. Substitution of conserved adenine nucleotides in mgtM abolished the response to ATP but had no effect in the response to proline limitation. This provides a singular example of a leader mRNA with tandem attenuators responding to different signals. PMID:22857388

  5. Isolation of Salmonella enterica subspecies enterica serovar Paratyphi B dT+, or Salmonella Java, from Indonesia and alteration of the d-tartrate fermentation phenotype by disrupting the ORF STM 3356.

    Science.gov (United States)

    Han, Kyung Ho; Choi, Seon Young; Lee, Je Hee; Lee, Hyejon; Shin, Eun Hee; Agtini, Magdarina D; von Seidlein, Lorenz; Ochiai, R Leon; Clemens, John D; Wain, John; Hahn, Ji-Sook; Lee, Bok Kwon; Song, Manki; Chun, Jongsik; Kim, Dong Wook

    2006-12-01

    Salmonella enterica subspecies enterica serovar Paratyphi B [O1,4,(5),12 : Hb : 1,2] can cause either an enteric fever (paratyphoid fever) or self-limiting gastroenteritis in humans. The d-tartrate non-fermenting variant S. enterica subsp. enterica serovar Paratyphi B dT- (S. Paratyphi B) is the causative agent of paratyphoid fever, and the d-tartrate fermenting variant S. enterica subsp. enterica serovar Paratyphi B dT+ (S. Paratyphi B dT+; formerly called Salmonella Java) causes gastroenteritis. S. Java is currently recognized as an emerging problem worldwide. Twelve dT+ S. Java isolates were collected in Indonesia between 2000 and 2002. One-third of them contained Salmonella genomic island 1 (SGI1), which gives the multidrug-resistant phenotype to the bacteria. In this study, a PCR-based method to detect a single nucleotide difference responsible for the inability to ferment d-tartrate, reported elsewhere, was validated. The d-tartrate fermenting phenotype of S. Java was converted to the non-fermenting phenotype by the disruption of the ORF STM 3356, and the d-tartrate non-fermenting phenotype of the ORF STM 3356-disrupted strain and the dT- reference strain was changed to the dT+ phenotype by complementing ORF STM 3356 in trans. The results show that the dT+ phenotype requires a functional product encoded by STM 3356, and support the use of the PCR-based discrimination method for S. Paratyphi B and S. Java as the standard differentiation method.

  6. Haloarchaeal gas vesicle nanoparticles displaying Salmonella SopB antigen reduce bacterial burden when administered with live attenuated bacteria.

    Science.gov (United States)

    DasSarma, Priya; Negi, Vidya Devi; Balakrishnan, Arjun; Karan, Ram; Barnes, Susan; Ekulona, Folasade; Chakravortty, Dipshikha; DasSarma, Shiladitya

    2014-07-31

    Innovative vaccines against typhoid and other Salmonella diseases that are safe, effective, and inexpensive are urgently needed. In order to address this need, buoyant, self-adjuvating gas vesicle nanoparticles (GVNPs) from the halophilic archaeon Halobacterium sp. NRC-1 were bioengineered to display the highly conserved Salmonella enterica antigen SopB, a secreted inosine phosphate effector protein injected by pathogenic bacteria during infection into the host cell. Two highly conserved sopB gene segments near the 3'-coding region, named sopB4 and B5, were each fused to the gvpC gene, and resulting GVNPs were purified by centrifugally accelerated flotation. Display of SopB4 and B5 antigenic epitopes on GVNPs was established by Western blotting analysis using antisera raised against short synthetic peptides of SopB. Immunostimulatory activities of the SopB4 and B5 nanoparticles were tested by intraperitoneal administration of recombinant GVNPs to BALB/c mice which had been immunized with S. enterica serovar Typhimurium 14028 ΔpmrG-HM-D (DV-STM-07), a live attenuated vaccine strain. Proinflammatory cytokines IFN-γ, IL-2, and IL-9 were significantly induced in mice boosted with SopB5-GVNPs, consistent with a robust Th1 response. After challenge with virulent S. enterica serovar Typhimurium 14028, bacterial burden was found to be diminished in spleen of mice boosted with SopB4-GVNPs and absent or significantly diminished in liver, mesenteric lymph node, and spleen of mice boosted with SopB5-GVNPs, indicating that the C-terminal portions of SopB displayed on GVNPs elicit a protective response to Salmonella infection in mice. SopB antigen-GVNPs were found to be stable at elevated temperatures for extended periods without refrigeration in Halobacterium cells. The results all together show that bioengineered GVNPs are likely to represent a valuable platform for the development of improved vaccines against Salmonella diseases.

  7. An oral recombinant Salmonella enterica serovar Typhimurium mutant elicits systemic antigen-specific CD8+ T cell cytokine responses in mice

    Directory of Open Access Journals (Sweden)

    Chin'ombe Nyasha

    2009-04-01

    Full Text Available Abstract Background The induction of antigen-specific CD8+ T cell cytokine responses against an attenuated, oral recombinant Salmonella enterica serovar Typhimurium vaccine expressing a green fluorescent protein (GFP model antigen was investigated. A GFP expression plasmid was constructed in which the gfp gene was fused in-frame with the 5' domain of the Escherichia coli β-galactosidase α-gene fragment with expression under the lac promoter. Groups of mice were orally immunized three times with the bacteria and systemic CD8+ T cell cytokine responses were evaluated. Results High level of the GFP model antigen was expressed by the recombinant Salmonella vaccine vector. Systemic GFP-specific CD8+ T cell cytokine (IFN-γ and IL-4 immune responses were detected after mice were orally vaccinated with the bacteria. It was shown that 226 net IFN-γ and 132 net IL-4 GFP-specific SFUs/10e6 splenocytes were formed in an ELISPOT assay. The level of IFN-γ produced by GFP peptide-stimulated cells was 65.2-fold above background (p Conclusion These results suggested that a high expressing recombinant Salmonella vaccine given orally to mice would elicit antigen-specific CD8+ T cell responses in the spleen. Salmonella bacteria may, therefore, be used as potential mucosal vaccine vectors.

  8. Prevención de la infección por Salmonella enterica subespecie enterica serotipo Enteritidis (Salmonella Enteritidis en pollos mediante un bacteriófago Prevention of Salmonella enterica subspecie enterica serotype Enteritidis (Salmonella Enteritidis infection in chickens using a bacteriophage

    Directory of Open Access Journals (Sweden)

    C Borie

    2008-01-01

    Full Text Available Las infecciones por Salmonella Enteritidis son una causa importante de enfermedades transmitidas por alimentos, epidemiológicamente asociada al consumo de productos derivados de las aves. Ya que el tratamiento con antimicrobianos causa la aparición de cepas multirresistentes, los fagos pueden ser usados como método alternativo para controlar S. Enteritidis en la industria avícola. El propósito de este trabajo fue evaluar el efecto del bacteriófago f3αSE sobre la incidencia de S. Enteritidis en pollos. Se formaron 5 grupos de 15 pollos broiler de 10 días de edad; los grupos experimentales A y B recibieron vía oral 1 ml del fago, conteniendo 10(6 y 10(7 UFP/dosis, respectivamente. Dos horas después, las aves fueron desafiadas con 1 mi de S. Enteritidis (4 x 10(6 UFC/dosis. El grupo C sólo recibió fagos (10(7 UFP/dosis y el grupo D sólo S. Enteritidis (4 x 10(6 UFC/dosis; el grupo E no recibió tratamiento y constituyó el grupo control sano. Diez días postinfección, los pollos recibieron eutanasia por inhalación de gas, obteniéndose por cada ave muestras de intestino y órganos para el reaislamiento de la cepa desafío y del fago. La incidencia de infección por S. Enteritidis se redujo en el grupo que recibió 10(7 UFP/dosis (p = 0,028 (7/15 aves del bacteriófago, no así en el grupo que recibió la dosis de 10(6 UFP (p = 0,061 (8/15 aves. La disminución de la incidencia de 5. Enteritidis en pollos, lograda mediante el fago f3αSE, permite considerarlo como una posible alternativa al uso de antimicrobianos en el control de Salmonella.Infections caused by Salmonella enterica subspecies enterica serotype Enteritidis (Salmonella Enteritidis are an important cause of foodborne diseases, epidemiologically associated with the consumption of poultry products. Since antibiotic treatments cause the appearance of multiresistant strains, phages can be used as an alternative method for controlling S. Enteritidis in the poultry industry

  9. Antimicrobial Resistance and Molecular Investigation of H2S-Negative Salmonella enterica subsp. enterica serovar Choleraesuis Isolates in China.

    Directory of Open Access Journals (Sweden)

    Jing Xie

    Full Text Available Salmonella enterica subsp. enterica serovar Choleraesuis is a highly invasive pathogen of swine that frequently causes serious outbreaks, in particular in Asia, and can also cause severe invasive disease in humans. In this study, 21 S. Choleraesuis isolates, detected from 21 patients with diarrhea in China between 2010 and 2011, were found to include 19 H2S-negative S. Choleraesuis isolates and two H2S-positive isolates. This is the first report of H2S-negative S. Choleraesuis isolated from humans. The majority of H2S-negative isolates exhibited high resistance to ampicillin, chloramphenicol, gentamicin, tetracycline, ticarcillin, and trimethoprim-sulfamethoxazole, but only six isolates were resistant to norfloxacin. In contrast, all of the isolates were sensitive to cephalosporins. Fifteen isolates were found to be multidrug resistant. In norfloxacin-resistant isolates, we detected mutations in the gyrA and parC genes and identified two new mutations in the parC gene. Pulsed-field gel electrophoresis (PFGE, multilocus sequence typing (MLST, and clustered regularly interspaced short palindromic repeat (CRISPR analysis were employed to investigate the genetic relatedness of H2S-negative and H2S-positive S. Choleraesuis isolates. PFGE revealed two groups, with all 19 H2S-negative S. Choleraesuis isolates belonging to Group I and H2S-positive isolates belonging to Group II. By MLST analysis, the H2S-negative isolates were all found to belong to ST68 and H2S-positive isolates belong to ST145. By CRISPR analysis, no significant differences in CRISPR 1 were detected; however, one H2S-negative isolate was found to contain three new spacers in CRISPR 2. All 19 H2S-negative isolates also possessed a frame-shift mutation at position 760 of phsA gene compared with H2S-positive isolates, which may be responsible for the H2S-negative phenotype. Moreover, the 19 H2S-negative isolates have similar PFGE patterns and same mutation site in the phsA gene, these

  10. Production of Autoinducer 2 in Salmonella enterica Serovar Thompson Contributes to Its Fitness in Chickens but Not on Cilantro Leaf Surfaces

    OpenAIRE

    Brandl, M. T.; Miller, W. G.; Bates, A. H.; Mandrell, R E

    2005-01-01

    Food-borne illness caused by Salmonella enterica has been linked traditionally to poultry products but is associated increasingly with fresh fruits and vegetables. We have investigated the role of the production of autoinducer 2 (AI-2) in the ability of S. enterica serovar Thompson to colonize the chicken intestine and the cilantro phyllosphere. A mutant of S. enterica serovar Thompson that is defective in AI-2 production was constructed by insertional mutagenesis of luxS. The population size...

  11. 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

    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...

  12. Differential Killing of Salmonella enterica Serovar Typhi by Antibodies Targeting Vi and Lipopolysaccharide O:9 Antigen.

    Science.gov (United States)

    Hart, Peter J; O'Shaughnessy, Colette M; Siggins, Matthew K; Bobat, Saeeda; Kingsley, Robert A; Goulding, David A; Crump, John A; Reyburn, Hugh; Micoli, Francesca; Dougan, Gordon; Cunningham, Adam F; MacLennan, Calman A

    2016-01-01

    Salmonella enterica serovar Typhi expresses a capsule of Vi polysaccharide, while most Salmonella serovars, including S. Enteritidis and S. Typhimurium, do not. Both S. Typhi and S. Enteritidis express the lipopolysaccharide O:9 antigen, yet there is little evidence of cross-protection from anti-O:9 antibodies. Vaccines based on Vi polysaccharide have efficacy against typhoid fever, indicating that antibodies against Vi confer protection. Here we investigate the role of Vi capsule and antibodies against Vi and O:9 in antibody-dependent complement- and phagocyte-mediated killing of Salmonella. Using isogenic Vi-expressing and non-Vi-expressing derivatives of S. Typhi and S. Typhimurium, we show that S. Typhi is inherently more sensitive to serum and blood than S. Typhimurium. Vi expression confers increased resistance to both complement- and phagocyte-mediated modalities of antibody-dependent killing in human blood. The Vi capsule is associated with reduced C3 and C5b-9 deposition, and decreased overall antibody binding to S. Typhi. However, purified human anti-Vi antibodies in the presence of complement are able to kill Vi-expressing Salmonella, while killing by anti-O:9 antibodies is inversely related to Vi expression. Human serum depleted of antibodies to antigens other than Vi retains the ability to kill Vi-expressing bacteria. Our findings support a protective role for Vi capsule in preventing complement and phagocyte killing of Salmonella that can be overcome by specific anti-Vi antibodies, but only to a limited extent by anti-O:9 antibodies.

  13. Differential Killing of Salmonella enterica Serovar Typhi by Antibodies Targeting Vi and Lipopolysaccharide O:9 Antigen.

    Directory of Open Access Journals (Sweden)

    Peter J Hart

    Full Text Available Salmonella enterica serovar Typhi expresses a capsule of Vi polysaccharide, while most Salmonella serovars, including S. Enteritidis and S. Typhimurium, do not. Both S. Typhi and S. Enteritidis express the lipopolysaccharide O:9 antigen, yet there is little evidence of cross-protection from anti-O:9 antibodies. Vaccines based on Vi polysaccharide have efficacy against typhoid fever, indicating that antibodies against Vi confer protection. Here we investigate the role of Vi capsule and antibodies against Vi and O:9 in antibody-dependent complement- and phagocyte-mediated killing of Salmonella. Using isogenic Vi-expressing and non-Vi-expressing derivatives of S. Typhi and S. Typhimurium, we show that S. Typhi is inherently more sensitive to serum and blood than S. Typhimurium. Vi expression confers increased resistance to both complement- and phagocyte-mediated modalities of antibody-dependent killing in human blood. The Vi capsule is associated with reduced C3 and C5b-9 deposition, and decreased overall antibody binding to S. Typhi. However, purified human anti-Vi antibodies in the presence of complement are able to kill Vi-expressing Salmonella, while killing by anti-O:9 antibodies is inversely related to Vi expression. Human serum depleted of antibodies to antigens other than Vi retains the ability to kill Vi-expressing bacteria. Our findings support a protective role for Vi capsule in preventing complement and phagocyte killing of Salmonella that can be overcome by specific anti-Vi antibodies, but only to a limited extent by anti-O:9 antibodies.

  14. Molecular typing of Salmonella enterica serovar typhi isolates from various countries in Asia by a multiplex PCR assay on variable-number tandem repeats.

    Science.gov (United States)

    Liu, Yichun; Lee, May-Ann; Ooi, Eng-Eong; Mavis, Yeo; Tan, Ai-Ling; Quek, Hung-Hiang

    2003-09-01

    A multiplex PCR method incorporating primers flanking three variable-number tandem repeat (VNTR) loci (arbitrarily labeled TR1, TR2, and TR3) in the CT18 strain of Salmonella enterica serovar Typhi has been developed for molecular typing of S. enterica serovar Typhi clinical isolates from several Asian countries, including Singapore, Indonesia, India, Bangladesh, Malaysia, and Nepal. We have demonstrated that the multiplex PCR could be performed on crude cell lysates and that the VNTR banding profiles produced could be easily analyzed by visual inspection after conventional agarose gel electrophoresis. The assay was highly discriminative in identifying 49 distinct VNTR profiles among 59 individual isolates. A high level of VNTR profile heterogeneity was observed in isolates from within the same country and among countries. These VNTR profiles remained stable after the strains were passaged extensively under routine laboratory culture conditions. In contrast to the S. enterica serovar Typhi isolates, an absence of TR3 amplicons and a lack of length polymorphisms in TR1 and TR2 amplicons were observed for other S. enterica serovars, such as Salmonella enterica serovar Typhimurium, Salmonella enterica serovar Enteritidis, and Salmonella enterica serovar Paratyphi A, B, and C. DNA sequencing of the amplified VNTR regions substantiated these results, suggesting the high stability of the multiplex PCR assay. The multiplex-PCR-based VNTR profiling developed in this study provides a simple, rapid, reproducible, and high-resolution molecular tool for the epidemiological analysis of S. enterica serovar Typhi strains. PMID:12958274

  15. 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.

  16. Antimicrobial activity of different copper alloy surfaces against copper resistant and sensitive Salmonella enterica.

    Science.gov (United States)

    Zhu, Libin; Elguindi, Jutta; Rensing, Christopher; Ravishankar, Sadhana

    2012-05-01

    Copper has shown antibacterial effects against foodborne pathogens. The objective of this study was to evaluate the antibacterial activity of copper surfaces on copper resistant and sensitive strains of Salmonella enterica. Six different copper alloy coupons (60-99.9% copper) were tested along with stainless steel as the control. The coupons were surface inoculated with either S. Enteritidis or one of the 3 copper resistant strains, S. Typhimurium S9, S19 and S20; stored under various incubation conditions at room temperature; and sampled at various times up to 2 h. The results showed that under dry incubation conditions, Salmonella only survived 10-15 min on high copper content alloys. Salmonella on low copper content alloys showed 3-4 log reductions. Under moist incubation conditions, no survivors were detected after 30 min-2 h on high copper content alloys, while the cell counts decreased 2-4 logs on low copper content coupons. Although the copper resistant strains survived better than S. Enteritidis, they were either completely inactivated or survival was decreased. Copper coupons showed better antimicrobial efficacy in the absence of organic compounds. These results clearly show the antibacterial effects of copper and its potential as an alternative to stainless steel for selected food contact surfaces.

  17. Development of ceftriaxone resistance affects the virulence properties of Salmonella enterica serotype Typhimurium strains.

    Science.gov (United States)

    Li, Liang; Yang, Yu-Rong; Liao, Xiao-Ping; Lei, Chun-Yin; Sun, Jian; Li, Lu-Lu; Liu, Bao-Tao; Yang, Shou-Shen; Liu, Ya-Hong

    2013-01-01

    Development of antibiotic resistance may alter the virulence properties of bacterial organisms. In this study, nine clinical ceftriaxone-susceptible Salmonella enterica serotype Typhimurium strains were subjected to stepwise selection with increasing concentrations of ceftriaxone in culture media. Mutations in virulence-associated genes and antibiotic efflux genes were analyzed by polymerase chain reaction (PCR) and DNA sequencing. The expression levels of virulence genes invA and stn as well as efflux pump genes tolC, arcA, and arcB before and after the selection were measured by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The stepwise selection resulted in the development of Salmonella strains that were highly resistant to ceftriaxone. Sequence analysis did not reveal any mutations or deletions in the examined virulence genes and regulatory gene, but a silent mutation (T423C) in acrR (encoding a repressor for the efflux pump) was detected in most of the ceftriaxone-resistant strains. The qRT-PCR revealed increased expression of the AcrAB-TolC efflux pump and decreased expression of invA and stn in the ceftriaxone-resistant strains. Moreover, decreased invasion into cultured epithelial cells and reduced growth rates were observed with the resistant strains. These results suggest that acquisition of ceftriaxone resistance is associated with the overexpression of the AcrAB-TolC efflux pump and leads to reduced virulence in Salmonella Typhimurium.

  18. Evaluation of nisin-β-lactam antibiotics against clinical strains of Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Rishi, Praveen; Preet Singh, Aman; Garg, Neha; Rishi, Madhuri

    2014-12-01

    There is an imperative need to discover novel antimicrobials and anti-infective agents and build up innovative strategies to combat multidrug-resistant Salmonella. In this context, we had earlier confirmed that nisin has the potential to act in conjunction with β-lactams against murine salmonellosis using standard strain. However, evaluation of efficacy of these combinations against clinical isolates of Salmonella could be the next key step to confirm the value added potential of this peptide. The present study was therefore planned to validate the synergistic effects of nisin-β-lactams combinations against clinical isolates of Salmonella enterica serovar Typhi. MICs of the selected β-lactams, EDTA and nisin were determined by micro and macro broth dilution assays. In-vitro synergism between the agents was evaluated by fractional inhibitory concentration (FIC) index (checkerboard test) and time-kill assay. All the tested combinations showed synergy against the tested clinical strains except three, as evident by FIC index (checkerboard test) and time-kill assay. Especially, nisin-ceftriaxone and nisin-cefotaxime combinations demonstrated excellent synergistic activity. In view of the encouraging results obtained from the previous and present study, further studies need to be carried out using large number of strains from different regions to take into account the clinical variability of the strains. Though it is a simple study but highlights an important point about the possibility of using the said combination while making strategies to develop newer formulations.

  19. Prevalence of enterobacteriaceae in Tupinambis merianae (Squamata: Teiidae from a captive facility in Central Brazil, with a profile of antimicrobial drug resistance in Salmonella enterica

    Directory of Open Access Journals (Sweden)

    Andréa de Moraes Carvalho

    2013-06-01

    Full Text Available The present study reports the presence of enterobacteriaceae in Tegu Lizards (Tupinambis merianaefrom a captive facility in central Brazil. From a total of 30 animals, 10 juveniles and 20 adults (10 males, 10 females, 60 samples were collected, in two periods separated by 15 days. The samples were cultivated in Xylose-lysine-deoxycholate agar (XLT4 and MacConkey agar. The Salmonella enterica were tested for antimicrobial susceptibility. A total of 78 bacteria was isolated, of wich 27 were from juveniles of T. merianae, 30 from adult males and 21 from adult females. Salmonella enterica was the most frequent bacteria followed by Citrobacter freundii, Escherichia coli, Enterobacter sakasakii, Kluivera sp., Citrobacter amalonaticus, Serratia marcescens, Citrobacter diversus, Yersinia frederiksenii, Serratia odorifera, and Serratia liquefaciens. Salmonella enterica subsp. diarizonae and houtenae showed resistance to cotrimoxazole, and serum Salmonella enterica Worthington showed resistance to tetracycline and gentamicin. Salmonella enterica Panama and S. enterica subsp. diarizonae showed intermediate sensitivity to cotrimoxazole. In addition to Enterobacteriaceae in the Tegu lizard, pathogenic serotypes of S. enterica also occur, and their antimicrobial resistance was confirmed.

  20. The role of litter beetles as potential reservoir for Salmonella enterica and thermophilic Campylobacter spp. between broiler flocks

    DEFF Research Database (Denmark)

    Skov, M.N.; Spencer, A.G.; Hald, Birthe;

    2004-01-01

    We evaluated the role of beetles infesting broiler chicken rearing facilities as potential reservoirs for Salmonella enterica infections between successive broiler flocks. In addition, their role as potential reservoirs for thermophilic Campylobacter spp. was also investigated. Fourteen broiler...... houses located at 11 different farms were included in the study. The houses were nonrandomly selected on the basis of their salmonella status; nine were persistently contaminated with salmonella whereas five were salmonella negative. For each broiler house, two consecutive broiler flocks (i.e., 28...... broiler flocks in all) as well as beetles collected during both rotations of production and in the empty period (after cleaning and disinfection) between these flocks were monitored for the presence of salmonella. Examinations for the presence of campylobacter in the same sample materials were also...

  1. Role of RpoS in fine-tuning the synthesis of Vi capsular polysaccharide in Salmonella enterica serotype Typhi.

    Science.gov (United States)

    Santander, Javier; Wanda, Soo-Young; Nickerson, Cheryl A; Curtiss, Roy

    2007-03-01

    Regulation of the synthesis of Vi polysaccharide, a major virulence determinant in Salmonella enterica serotype Typhi, is under the control of two regulatory systems, ompR-envZ and rscB-rscC, which respond to changes in osmolarity. Some serotype Typhi strains exhibit overexpression of Vi polysaccharide, which masks clinical detection of lipopolysaccharide O antigen. This variation in Vi polysaccharide and O antigen display (VW variation) has been observed since the initial studies of serotype Typhi. In this study, we report that rpoS plays a role in this increased expression in Vi polysaccharide. We constructed a variety of isogenic serotype Typhi mutants that differed in their expression levels of RpoS and examined the role of the rpoS product in synthesis of Vi polysaccharide under different osmolarity conditions. Vi polysaccharide synthesis was also examined in serotype Typhi mutants in which the native promoter of the rpoS was replaced by an araCP(BAD) cassette, so that the expression of rpoS was arabinose dependent. The RpoS(-) strains showed increased syntheses of Vi polysaccharide, which at low and medium osmolarities masked O antigen detection. In contrast, RpoS(+) strains showed lower syntheses of Vi polysaccharide, and an increased detection of O antigen was observed. During exponential growth, when rpoS is unstable or present at low levels, serotype Typhi RpoS(+) strains overexpress the Vi polysaccharide at levels comparable to those for RpoS(-) strains. Our results show that RpoS is another regulator of Vi polysaccharide synthesis and contributes to VW variation in serotype Typhi, which has implications for the development of recombinant attenuated Salmonella vaccines in humans.

  2. Effect of antimicrobial exposure on AcrAB expression in Salmonella enterica subspecies enterica serovar Choleraesuis

    Directory of Open Access Journals (Sweden)

    Masaru eUsui

    2013-03-01

    Full Text Available Understanding the impact of antimicrobial use on the emergence of resistant bacteria is imperative to prevent its emergence. For instance, activation of the AcrAB efflux pumps is responsible for the emergence of antimicrobial-resistant Salmonella strains. Here, we examined the expression levels of acrB and its multiple regulator genes (RamA, SoxS, MarA, and Rob in 17 field isolates of S. Choleraesuis by using quantitative PCR methods. The expression of acrB increased in 8 of the field isolates (P < 0.05. The expression of acrB was associated with that of ramA in 1 isolate, soxS in 1 isolate, and both these genes in 6 isolates. Thereafter, to examine the effect of selected antimicrobials (enrofloxacin, ampicillin, oxytetracycline, kanamycin, and spectinomycin on the expression of acrB and its regulator genes, mutants derived from 5 isolates of S. Choleraesuis were selected by culture on antimicrobial-containing plates. The expression of acrB and ramA was higher in the mutants selected using enrofloxacin (3.3–6.3- and 24.5–37.7-fold, respectively, ampicillin (1.8–7.7- and 16.1–55.9-fold, respectively, oxytetracycline (1.7–3.3- and 3.2–31.1-fold, respectively, and kanamycin (1.6–2.2- and 5.6–26.4-fold, respectively, which are AcrAB substrates, than in each of the parental strains (P < 0.05. In contrast, in AcrAB substrate-selected mutants, the expression of soxS, marA, and rob remained similar to that in parental strains. Of the 4 antimicrobials, the level of ramA expression was significantly higher in the enrofloxacin- and ampicillin-selected mutants than in the oxytetracycline- and kanamycin-selected mutants (P < 0.05, whereas the expression levels of acrB and multiple regulator genes in spectinomycin-selected mutants were similar to those in each parental strain. These data suggest that exposure to antimicrobials that are AcrAB substrates enhance the activation of the AcrAB efflux pump via RamA, but not via SoxS, MarA, or Rob in

  3. Comparison of a PCR serotyping assay, Check&Trace assay for Salmonella, and Luminex Salmonella serotyping assay for the characterization of Salmonella enterica identified from fresh and naturally contaminated cilantro.

    Science.gov (United States)

    Jean-Gilles Beaubrun, J; Ewing, L; Jarvis, K; Dudley, K; Grim, C; Gopinath, G; Flamer, M-L; Auguste, W; Jayaram, A; Elmore, J; Lamont, M; McGrath, T; Hanes, D E

    2014-09-01

    Salmonella enterica isolated from fresh cilantro samples collected through the USDA/AMS Microbiological Data Program (MDP) were used to compare a PCR serotyping assay against the Check&Trace assay and the Luminex (BioPlex) Salmonella serotyping assay. The study was conducted to evaluate the effectiveness of the three methods for serotyping Salmonella from both enrichment broth cultures and pure Salmonella cultures. In this investigation, Salmonella spp. serotyping was conducted using 24 h enrichment broth cultures and pure Salmonella cultures from cilantro samples, with the PCR serotyping assay. Conversely, the Check&Trace and Luminex for Salmonella assays required pure cultures for Salmonella serotyping. The cilantro samples contained S. enterica serovar Montevideo, Newport, Saintpaul, and Tennessee, identified by the PCR serotyping assay and Check&Trace for Salmonella, but the Luminex assay only identified two of the four serotypes of the cilantro samples. The anticipated impact from this study is that the PCR serotyping assay provides a time- and cost-effective means for screening, identifying and serotyping Salmonella using DNA extracted from 24 h enrichment cilantro samples. PMID:24929735

  4. Construcción de mutantes de Salmonella enterica por inactivación de los genes invG/invE y ssaJ/ssaK de las islas de patogenicidad 1 y 2

    Directory of Open Access Journals (Sweden)

    Judith Velasco

    2010-10-01

    To understand the genetic basis of Salmonella pathogenicity mechanisms, various methods have been described to manipulate and generate mutant genomic DNA with specific characteristics. In this study we report the construction of mutants from several serotypes of S. enterica, substitution and inactivation of genes invG/invE in SPI-1 gene and ssaJ/ssaK in SPI-2 by the technique of phage λ Red recombinase, as described by Datsenko and Wanner (2000. The gene deletion in SPI-1 and SPI-2 encodes proteins involved in the formation of type III secretion systems responsible for the invasion and intracellular survival of S. enterica in the host cells. The results of this work will allow in vivo studies to evaluate the possible attenuation of virulence of the mutant strains, as well as to provide new insights into the genetic mechanisms involved in the pathogenesis of diseases caused by these bacteria. Moreover, this technique is recommended to efficiently generate mutants of different serotypes of S. enterica in order to study the chromosomal genes and their products. Key words: red recombinasa, mutation, pathogenicity islands, Salmonella enterica.

  5. Development of stable reporter system cloning luxCDABE genes into chromosome of Salmonella enterica serotypes using Tn7 transposon

    Directory of Open Access Journals (Sweden)

    Lawrence Mark L

    2010-07-01

    Full Text Available Abstract Background Salmonellosis may be a food safety problem when raw food products are mishandled and not fully cooked. In previous work, we developed bioluminescent Salmonella enterica serotypes using a plasmid-based reporting system that can be used for real-time monitoring of the pathogen's growth on food products in short term studies. In this study, we report the use of a Tn7-based transposon system for subcloning of luxCDABE genes into the chromosome of eleven Salmonella enterica serotypes isolated from the broiler production continuum. Results We found that the lux operon is constitutively expressed from the chromosome post-transposition and the lux cassette is stable without external pressure, i.e. antibiotic selection, for all Salmonella enterica serotypes used. Bioluminescence expression is based on an active electron transport chain and is directly related with metabolic activity. This relationship was quantified by measuring bioluminescence against a temperature gradient in aqueous solution using a luminometer. In addition, bioluminescent monitoring of two serotypes confirmed that our chicken skin model has the potential to be used to evaluate pathogen mitigation strategies. Conclusions This study demonstrated that our new stable reporting system eliminates bioluminescence variation due to plasmid instability and provides a reliable real-time experimental system to study application of preventive measures for Salmonella on food products in real-time for both short and long term studies.

  6. 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.

  7. 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. PMID:27357046

  8. 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

    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......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...... of the hatcheries, and with five houses on the farm. An interaction between season anal the previously mentioned hatcheries, and a random effect at farm level was also found to be statistically significant. Twelve variables were not found to be associated with S, typhimurium infection: medication, growth promoters...

  9. Evaluation of whole genome sequencing for outbreak detection of Salmonella enterica

    DEFF Research Database (Denmark)

    Leekitcharoenphon, Pimlapas; Nielsen, Eva M.; Kaas, Rolf Sommer;

    2014-01-01

    Salmonella enterica is a common cause of minor and large food borne outbreaks. To achieve successful and nearly ‘real-time’ monitoring and identification of outbreaks, reliable sub-typing is essential. Whole genome sequencing (WGS) shows great promises for using as a routine epidemiological typing...... tool. Here we evaluate WGS for typing of S. Typhimurium including different approaches for analyzing and comparing the data. A collection of 34 S. Typhimurium isolates was sequenced. This consisted of 18 isolates from six outbreaks and 16 epidemiologically unrelated background strains. In addition, 8 S....... Enteritidis and 5 S. Derby were also sequenced and used for comparison. A number of different bioinformatics approaches were applied on the data; including pan-genome tree, k-mer tree, nucleotide difference tree and SNP tree. The outcome of each approach was evaluated in relation to the association...

  10. An optimized affordable DNA-extraction method from Salmonella enterica Enteritidis for PCR experiments

    Directory of Open Access Journals (Sweden)

    Karimnasab, N.,

    2013-12-01

    Full Text Available In diagnostic and research bacteriology settings with budget and staff restrictions, fast and cost-effective genome extraction methods are desirable. If not inactivated properly, cellular and/or environmental DNA nucleases will degrade genomic material during the extraction stage, and therefore might give rise to incorrect results in PCR experiments. When crude cell extracts, proteinase K–treated templates and purified DNAs prepared by phenol-chloroform-isoamylalcohol method as well as a commercial extraction kit were subjected to the Salmonella enterica Enteritidis specific STM2 PCR, with exception of crude cell extract, PCR products from all other three methods saved their integrity for 28 days post-generation. This work aimed to find out whether improvement to boiling method can guaranty stability of PCR products. As results showed, treatment of crude cell extracts from S. Enteritidis with proteinase K offers an inexpensive, fast and effective DNA extraction method suitable for high-throughput laboratories.

  11. Distribution and Genetic Diversity of Salmonella enterica in the Upper Suwannee River

    Directory of Open Access Journals (Sweden)

    Masoumeh Rajabi

    2011-01-01

    Full Text Available The Suwannee River spans the Florida/Georgia border to the Gulf of Mexico, and contributes to regional irrigation and recreational activities. Association of Salmonella enterica with these resources may result in the contamination of produce and disease outbreaks. Therefore, surface water was examined for the distribution of S. enterica at multiple time points from 4 sites on the upper Suwannee River. Isolates were confirmed by detection of the invA gene, and 96% of all samples were positive for the bacterium. Most probable number enumeration ranged from 60% similarity and distributed into 16 rep-PCR genogroups. Most (74% of the Suwannee River isolates were clustered into two genogroups that were comprised almost exclusively (97% of just these isolates. Conversely, 85% of the clinical reference strains clustered into other genogroups. However, some Suwannee River isolates (12% were clustered with these primarily clinically-associated genogroups, supporting the hypothesis that river water can serve as a disease reservoir and that pathogenic strains may persist or possibly originate from environmental sources.

  12. Regulation of fucose and 1,2-propanediol utilization by Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Staib, Lena; Fuchs, Thilo M

    2015-01-01

    After ingestion, Salmonella enterica serovar Typhimurium (S. Typhimurium) encounters a densely populated, competitive environment in the gastrointestinal tract. To escape nutrient limitation caused by the intestinal microbiota, this pathogen has acquired specific metabolic traits to use compounds that are not metabolized by the commensal bacteria. For example, the utilization of 1,2-propanediol (1,2-PD), a product of the fermentation of L-fucose, which is present in foods of herbal origin and is also a terminal sugar of gut mucins. Under anaerobic conditions and in the presence of tetrathionate, 1,2-PD can serve as an energy source for S. Typhimurium. Comprehensive database analysis revealed that the 1,2-PD and fucose utilization operons are present in all S. enterica serovars sequenced thus far. The operon, consisting of 21 genes, is expressed as a single polycistronic mRNA. As demonstrated here, 1,2-PD was formed and further used when S. Typhimurium strain 14028 was grown with L-fucose, and the gene fucA encoding L-fuculose-1-phosphate aldolase was required for this growth. Using promoter fusions, we monitored the expression of the propanediol utilization operon that was induced at very low concentrations of 1,2-PD and was inhibited by the presence of D-glucose.

  13. Suppressor analyses identify threonine as a modulator of ridA mutant phenotypes in Salmonella enterica.

    Directory of Open Access Journals (Sweden)

    Melissa R Christopherson

    Full Text Available The RidA (YjgF/YER057c/UK114 family of proteins is broadly conserved in the three domains of life yet the functional understanding of these proteins is at an early stage. Physiological studies of ridA mutant strains of Salmonella enterica provided a framework to inform in vitro studies and led to the description of a conserved biochemical activity for this family. ridA mutant strains of S. enterica have characteristic phenotypes including new synthesis of thiamine biosynthetic intermediate phosphoribosylamine (PRA, inability to grow on pyruvate as a sole carbon and energy source or when serine is present in the minimal growth medium, and a decreased specific activity of transaminase B (IlvE. Secondary mutations restoring growth to a ridA mutant in the presence of serine were in dapA (encoding dihydrodipicolinate synthase and thrA (encoding homoserine dehydrogenase. These mutations suppressed multiple ridA mutant phenotypes by increasing the synthesis of threonine. The ability of threonine to suppress the metabolic defects of a ridA mutant is discussed in the context of recent biochemical data and in vivo results presented here.

  14. Salmonella Enterica Serovar Typhimurium BipA Exhibits Two Distinct Ribosome Binding Modes

    Energy Technology Data Exchange (ETDEWEB)

    deLivron, M.; Robinson, V

    2008-01-01

    BipA is a highly conserved prokaryotic GTPase that functions to influence numerous cellular processes in bacteria. In Escherichia coli and Salmonella enterica serovar Typhimurium, BipA has been implicated in controlling bacterial motility, modulating attachment and effacement processes, and upregulating the expression of virulence genes and is also responsible for avoidance of host defense mechanisms. In addition, BipA is thought to be involved in bacterial stress responses, such as those associated with virulence, temperature, and symbiosis. Thus, BipA is necessary for securing bacterial survival and successful invasion of the host. Steady-state kinetic analysis and pelleting assays were used to assess the GTPase and ribosome-binding properties of S. enterica BipA. Under normal bacterial growth, BipA associates with the ribosome in the GTP-bound state. However, using sucrose density gradients, we demonstrate that the association of BipA and the ribosome is altered under stress conditions in bacteria similar to those experienced during virulence. The data show that this differential binding is brought about by the presence of ppGpp, an alarmone that signals the onset of stress-related events in bacteria.

  15. Involvement of SPI-2-encoded SpiC in flagellum synthesis in Salmonella enterica serovar Typhimurium

    Directory of Open Access Journals (Sweden)

    Sugita Asami

    2009-08-01

    Full Text Available Abstract Background SpiC encoded within Salmonella pathogenicity island 2 on the Salmonella enterica serovar Typhimurium chromosome is required for survival within macrophages and systemic infection in mice. Additionally, SpiC contributes to Salmonella-induced activation of the signal transduction pathways in macrophages by affecting the expression of FliC, a component of flagella filaments. Here, we show the contribution of SpiC in flagellum synthesis. Results Quantitative RT-PCR shows that the expression levels of the class 3 fliD and motA genes that encode for the flagella cap and motor torque proteins, respectively, were lower for a spiC mutant strain than for the wild-type Salmonella. Further, this mutant had lower expression levels of the class 2 genes including the fliA gene encoding the flagellar-specific alternative sigma factor. We also found differences in flagella assembly between the wild-type strain and the spiC mutant. Many flagella filaments were observed on the bacterial surface of the wild-type strain, whereas the spiC mutant had only few flagella. The absence of spiC led to reduced expression of the FlhD protein, which functions as the master regulator in flagella gene expression, although no significant difference at the transcription level of the flhDC operon was observed between the wild-type strain and the spiC mutant. Conclusion The data show that SpiC is involved in flagella assembly by affecting the post-transcription expression of flhDC.

  16. 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. PMID:26325598

  17. Protection of epithelial cells from Salmonella enterica serovar Enteritidis invasion by antibodies against the SPI-1 type III secretion system.

    Science.gov (United States)

    Desin, Taseen S; Mickael, Claudia S; Lam, Po-King S; Potter, Andrew A; Köster, Wolfgang

    2010-06-01

    Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) is one of the major causes of bacterial food-borne illness in humans. During the course of infection, Salmonella Enteritidis uses 2 type III secretion systems (T3SS), one of which is encoded on Salmonella pathogenicity island 1 (SPI-1). SPI-1 plays a major role in the invasion process. In the present study, we evaluated the effect of sera against the SPI-1 T3SS components on invasion in vitro using polarized human intestinal epithelial cells (Caco-2). Antisera to SipD protected Caco-2 cells against entry of wild-type Salmonella Enteritidis. On the other hand, sera against InvG, PrgI, SipA, SipC, SopB, SopE, and SopE2 did not affect Salmonella Enteritidis entry. To illustrate the specificity of anti-SipD mediated inhibition, SipD-specific antibodies were depleted from the serum. Antiserum depleted of SipD-specific antibodies lost its capacity to inhibit Salmonella Enteritidis entry. Thus, we demonstrate for the first time that antibodies against the SPI-1 needle tip protein (SipD) inhibit Salmonella Enteritidis invasion and that the SipD protein may be an important target in blocking SPI-1 mediated virulence of Salmonella Enteritidis.

  18. Genomic relationships between selected phage types of Salmonella enterica subsp. enterica serotype typhimurium defined by ribotyping, IS200 typing and PFGE

    DEFF Research Database (Denmark)

    Olsen, J. E.; Skov, M. N.; Angen, Øystein;

    1997-01-01

    — DTs 69, 103 and 153. The same grouping was observed by principal component analysis, but a minimum spanning tree linked DT 12 to group E and not group D in this analysis. Among the typing methods used, IS200 gave the best representation of the overall similarity between the S. typhimurium isolates......The genomic relationship between isolates representing 17 definitive phage types (DTs) of Salmonella enterica subsp. enterica serotype typhimurium (S. typhimurium) were analysed using three different typing methods: IS200 typing using the restriction enzymes EcoRI and Pvull, ribotyping using Smal...... similarity was indicated between isolates of the following phage types: group A — DTs 44, 49, 135 and 204c, with DT 9 distantly related; group B — DTs 95 and 99; and group C — DTs 104a, 110 and 120. The other large cluster contained group D — DTs 10, 20 and 146, with DT 12 distantly related, and group E...

  19. Effects of leachate from crumb rubber and zinc in green roofs on the survival, growth, and resistance characteristics of Salmonella enterica subsp. enterica serovar Typhimurium.

    Science.gov (United States)

    Crampton, Mollee; Ryan, Allayna; Eckert, Cori; Baker, Katherine H; Herson, Diane S

    2014-05-01

    The use of green roofs is a growing practice worldwide, particularly in densely populated areas. In an attempt to find new methods for recycling crumb rubber, incorporation of crumb rubber into artificial medium for plant growth in green roofs and similar engineered environments has become an attractive option for the recycling of waste tires. Though this approach decreases waste in landfills, there are concerns about the leaching of zinc and other heavy metals, as well as nutrient and organic compounds, into the environment. The present study analyzed the impact of leachate from crumb rubber and zinc on the growth and viability of Salmonella enterica subsp. enterica serovar Typhimurium. Zinc was chosen for further studies since it has been previously implicated with other biological functions, including biofilm formation, motility, and possible cross-resistance to antimicrobial agents. The study showed that Salmonella can colonize crumb rubber and that crumb rubber extract may provide nutrients that are usable by this bacterium. Salmonella strains with reduced susceptibility (SRS) to zinc were obtained after subculturing in increasing concentrations of zinc. The SRS exhibited differences in gene expression of flux pump genes zntA and znuA compared to that of the parent when exposed to 20 mM added zinc. In biofilm formation studies, the SRS formed less biofilm but was more motile than the parental strain.

  20. Identification of bapA in Strains of Salmonella enterica subsp. enterica Isolated from Wild Animals Kept in Captivity in Sinaloa, Mexico.

    Science.gov (United States)

    Silva-Hidalgo, Gabriela; López-Valenzuela, Martin; Cárcamo-Aréchiga, Nora; Cota-Guajardo, Silvia; López-Salazar, Mayra; Montiel-Vázquez, Edith

    2016-01-01

    bapA, previously named stm2689, encodes the BapA protein, which, along with cellulose and fimbriae, constitutes biofilms. Biofilms are communities of microorganisms that grow in a matrix of exopolysaccharides and may adhere to living tissues or inert surfaces. Biofilm formation is associated with the ability to persist in different environments, which contributes to the pathogenicity of several species. We analyzed the presence of bapA in 83 strains belonging to 17 serovars of Salmonella enterica subsp. enterica from wildlife in captivity at Culiacan's Zoo and Mazatlán's Aquarium. Each isolate amplified a product of 667 bp, which corresponds to the expected size of the bapA initiator, with no observed variation between different serovars analyzed. bapA gene was found to be highly conserved in Salmonella and can be targeted for the genus-specific detection of this organism from different sources. Since bapA expression improves bacterial proliferation outside of the host and facilitates resistance to disinfectants and desiccation, the survival of Salmonella in natural habitats may be favored. Thus, the risk of bacterial contamination from these animals is increased. PMID:27379195

  1. Identification of bapA in Strains of Salmonella enterica subsp. enterica Isolated from Wild Animals Kept in Captivity in Sinaloa, Mexico

    Science.gov (United States)

    López-Valenzuela, Martin; Cárcamo-Aréchiga, Nora; Cota-Guajardo, Silvia; López-Salazar, Mayra; Montiel-Vázquez, Edith

    2016-01-01

    bapA, previously named stm2689, encodes the BapA protein, which, along with cellulose and fimbriae, constitutes biofilms. Biofilms are communities of microorganisms that grow in a matrix of exopolysaccharides and may adhere to living tissues or inert surfaces. Biofilm formation is associated with the ability to persist in different environments, which contributes to the pathogenicity of several species. We analyzed the presence of bapA in 83 strains belonging to 17 serovars of Salmonella enterica subsp. enterica from wildlife in captivity at Culiacan's Zoo and Mazatlán's Aquarium. Each isolate amplified a product of 667 bp, which corresponds to the expected size of the bapA initiator, with no observed variation between different serovars analyzed. bapA gene was found to be highly conserved in Salmonella and can be targeted for the genus-specific detection of this organism from different sources. Since bapA expression improves bacterial proliferation outside of the host and facilitates resistance to disinfectants and desiccation, the survival of Salmonella in natural habitats may be favored. Thus, the risk of bacterial contamination from these animals is increased. PMID:27379195

  2. Effects of leachate from crumb rubber and zinc in green roofs on the survival, growth, and resistance characteristics of Salmonella enterica subsp. enterica serovar Typhimurium.

    Science.gov (United States)

    Crampton, Mollee; Ryan, Allayna; Eckert, Cori; Baker, Katherine H; Herson, Diane S

    2014-05-01

    The use of green roofs is a growing practice worldwide, particularly in densely populated areas. In an attempt to find new methods for recycling crumb rubber, incorporation of crumb rubber into artificial medium for plant growth in green roofs and similar engineered environments has become an attractive option for the recycling of waste tires. Though this approach decreases waste in landfills, there are concerns about the leaching of zinc and other heavy metals, as well as nutrient and organic compounds, into the environment. The present study analyzed the impact of leachate from crumb rubber and zinc on the growth and viability of Salmonella enterica subsp. enterica serovar Typhimurium. Zinc was chosen for further studies since it has been previously implicated with other biological functions, including biofilm formation, motility, and possible cross-resistance to antimicrobial agents. The study showed that Salmonella can colonize crumb rubber and that crumb rubber extract may provide nutrients that are usable by this bacterium. Salmonella strains with reduced susceptibility (SRS) to zinc were obtained after subculturing in increasing concentrations of zinc. The SRS exhibited differences in gene expression of flux pump genes zntA and znuA compared to that of the parent when exposed to 20 mM added zinc. In biofilm formation studies, the SRS formed less biofilm but was more motile than the parental strain. PMID:24584242

  3. Incidence and growth of Salmonella enterica on the peel and pulp of avocado (Persea americana) and custard apple (Annona squamosa).

    Science.gov (United States)

    Rezende, Ana Carolina B; Crucello, Juliana; Moreira, Rafael C; Silva, Beatriz S; Sant'Ana, Anderson S

    2016-10-17

    The aim of this study was to assess the incidence and to estimate the growth kinetic parameters (maximum growth rate, μ; lag time, λ; and maximum population, κ) of Salmonella on the peel and pulp of avocado (Perseaamericana var. americana) and custard apple (Annona squamosa L.) as affected by temperature (10-30°C). The incidence of Salmonella was assessed on the peel and pulp of the fruits (n=200 of each fruit), separately, totalizing 800 analyses. Only three samples of custard apple pulp were positive for Salmonella enterica and the three isolates recovered belonged to serotype S. Typhimurium. Salmonella was not recovered from avocado and custard apple peels and from avocado pulp. Generally, the substrate (pulp or peel) of growth did not affect μ values of S. enterica (p>0.05). Very similar μ values were found for S. enterica inoculated in custard apple and avocado. S. enterica presented the highest λ in the peel of the fruits. The growth of S. enterica resulted in larger λ in custard apple in comparison to avocado. For example, the λ of S. enterica in the pulp of custard apple and avocado were 47.0±0.78h and 10.0±3.78h, respectively. The lowest values of κ were obtained at the lower storage temperature conditions (10°C). For instance, κ values of 3.7±0.06log CFU/g and 2.9±0.03log CFU/g were obtained from the growth of S. enterica in avocado and custard apple pulps at 10°C (p<0.05), respectively. On the other hand, at 30°C, κ values were 6.5±0.25log CFU/g and 6.5±0.05log CFU/g, respectively. Significantly higher κ were obtained from the growth of S. enterica in the pulp than in the peel of the fruits (p<0.05). For instance, the growth of S. enterica in the pulp of avocado led to a κ value of 6.5±0.25log CFU/g, while in the peel led to a κ value of 4.6±0.23log CFU/g (p<0.05). In general, growth kinetic parameters indicated that avocado comprises a better substrate than custard apple for the growth of S. enterica. The square root model

  4. Antibiotic resistances in Listeria monocytogenes and Salmonella enterica isolated from foods with animal origin Resistencias a antibióticos en Listeria monocytogenes y Salmonella enterica aislados de alimentos de origen animal

    OpenAIRE

    Baltasar Balsalobre Hernández; Joaquín Hernández-Godoy

    2004-01-01

    Extensive use of antibiotics in both human and animal health and in cattle production has generated resistant microorganisms to common antibiotics. Resistances spread caused by human and animal therapeutic is well known, but we know poorly frecuency of resistant bacteria in foods with animal origin and destinated to human consumers. In this paper, sensitivity to nineteen antibiotics was investigated in Listeria monocytogenes and Salmonella enterica strains isolated from foods with animal orig...

  5. Pathoadaptive mutations in Salmonella enterica isolated after serial passage in mice.

    Directory of Open Access Journals (Sweden)

    Sanna Koskiniemi

    Full Text Available How pathogenic bacteria adapt and evolve in the complex and variable environment of the host remains a largely unresolved question. Here we have used whole genome sequencing of Salmonella enterica serovar Typhimurium LT2 populations serially passaged in mice to identify mutations that adapt bacteria to systemic growth in mice. We found unique pathoadaptive mutations in two global regulators, phoQ and stpA, which increase the competitive indexes of the bacteria 3- to 5-fold. Also, all mouse-adapted lineages had changed the orientation of the hin invertable element, resulting in production of a FliC type of flagellum. Competition experiments in mice with locked flagellum mutants showed that strains expressing the FliC type of flagellum had a 5-fold increase in competitive index as compared to those expressing FljB type flagellum. Combination of the flagellum cassette inversion with the stpA mutation increased competitive indexes up to 20-fold. These experiments show that Salmonella can rapidly adapt to a mouse environment by acquiring a few mutations of moderate individual effect that when combined confer substantial increases in growth.

  6. 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.

  7. Pulsed field gel electrophoresis identifies an outbreak of Salmonella enterica serotype Montevideo infection associated with a supermarket hot food outlet.

    Science.gov (United States)

    Threlfall, E J; Hampton, M D; Ward, L R; Richardson, I R; Lanser, S; Greener, T

    1999-09-01

    In February 1996 Salmonella enterica serotype Montevideo infection in a patient in the North Tyneside area was attributed to consumption of cooked chicken bought from a supermarket hot food outlet. Isolates from the patient, leftover food, and environmental samples were indistinguishable by pulsed field gel electrophoresis (PFGE). PFGE also demonstrated that an outbreak of infection with S. Montevideo associated with the hot food outlet had occurred in late 1995 and early 1996. This study shows the importance of microbial strain discrimination in outbreak investigations and illustrates the value of close liaison between microbiologists, epidemiologists, and environmental health officers in the control of salmonella outbreaks.

  8. Genomic and Phenotypic Analyses Reveal the Emergence of an Atypical Salmonella enterica Serovar Senftenberg Variant in China

    KAUST Repository

    Abd El Ghany, Moataz

    2016-05-25

    Human infections with Salmonella enterica subspecies enterica serovar Senftenberg are often associated with exposure to poultry flocks, farm environments, or contaminated food. The recent emergence of multidrug-resistant isolates has raised public health concerns. In this study, comparative genomics and phenotypic analysis were used to characterize 14 Salmonella Senftenberg clinical isolates recovered from multiple outbreaks in Shenzhen and Shanghai, China, between 2002 and 2011. Single-nucleotide polymorphism analyses identified two phylogenetically distinct clades of S. Senftenberg, designated SC1 and SC2, harboring variations in Salmonella pathogenicity island 1 (SPI-1) and SPI-2 and exhibiting distinct biochemical and phenotypic signatures. Although the two variants shared the same serotype, the SC2 isolates of sequence type 14 (ST14) harbored intact SPI-1 and -2 and hence were characterized by possessing efficient invasion capabilities. In contrast, the SC1 isolates had structural deletion patterns in both SPI-1 and -2 that correlated with an impaired capacity to invade cultured human cells and also the year of their isolation. These atypical SC1 isolates also lacked the capacity to produce hydrogen sulfide. These findings highlight the emergence of atypical Salmonella Senftenberg variants in China and provide genetic validation that variants lacking SPI-1 and regions of SPI-2, which leads to impaired invasion capacity, can still cause clinical disease. These data have identified an emerging public health concern and highlight the need to strengthen surveillance to detect the prevalence and transmission of nontyphoidal Salmonella species.

  9. A third mode of surface‐associated growth: immobilization of Salmonella enterica serovar Typhimurium modulates the RpoS‐directed transcriptional programme

    DEFF Research Database (Denmark)

    Knudsen, Gitte Maegaard; Nielsen, Maj‐Britt; Grassby, Terri;

    2012-01-01

    of Salmonella enterica serovar Typhimurium ST4/74 were compared during planktonic and immobilized growth, and a number of immobilization‐specific characteristics were identified. Immobilized S. Typhimurium did not express motility and chemotaxis genes, and electron microscopy revealed the absence of flagella...... that is distinct from the biofilm and swarming lifestyles of Salmonella....

  10. Identification of novel Salmonella enterica Serovar Thyphimurium DT104-specific prophage and nonprophage chromosomal sequences among serovar Thyphimurium isolates by genomic subtractive hybridization

    NARCIS (Netherlands)

    Hermans, A.P.H.M.; Abee, T.; Zwietering, M.H.; Aarts, H.J.M.

    2005-01-01

    Genomic subtractive hybridization was performed between Salmonella enterica serovar Typhimurium LT2 and DT104 to search for novel Salmonella serovar Typhimurium DT104-specific sequences. The subtraction resulted mainly in the isolation of DNA fragments with sequence similarity to phages. Two fragmen

  11. Complete Genome Sequence of Salmonella enterica Serovar Typhimurium Strain SO3 (Sequence Type 302) Isolated from a Baby with Meningitis in Mexico

    Science.gov (United States)

    Puente, José L.; Calva, Edmundo; Zaidi, Mussaret B.

    2016-01-01

    The complete genome of Salmonella enterica serovar Typhimurium strain SO3 (sequence type 302), isolated from a fatal meningitis infection in Mexico, was determined using PacBio technology. The chromosome hosts six complete prophages and is predicted to harbor 51 genomic islands, including 13 pathogenicity islands (SPIs). It carries the Salmonella virulence plasmid (pSTV). PMID:27103717

  12. Antimicrobial resistance genes in multidrug-resistant Salmonella enterica isolated from animals, retail meats, and humans in the United States and Canada

    Science.gov (United States)

    Salmonella enterica is a prevalent foodborne pathogen which can carry multi-drug resistance (MDR) and pose a threat to human health. Identifying the genetic elements associated with MDR in Salmonella isolated from animals, foods, and humans can help determine the sources of MDR in food animals and t...

  13. Comparison of API 20E and invA PCR for identification of Salmonella enterica isolates from swine production units.

    Science.gov (United States)

    Nucera, Daniele M; Maddox, Carol W; Hoien-Dalen, Patricia; Weigel, Ronald M

    2006-09-01

    API 20E and invA PCR were evaluated for the identification of Salmonella enterica isolates from swine farms. API 20E had the highest agreement with other tests at the 99.9% likelihood level. Both tests had 100% sensitivity and 96% specificity compared to 16S rRNA sequencing. Compared to serotyping, both tests had 96% sensitivity; specificity was 86% for API 20E and 79% for invA PCR.

  14. ramR mutations affecting fluoroquinolone susceptibility in epidemic multidrug-resistant Salmonella enterica serovar Kentucky ST198

    OpenAIRE

    2013-01-01

    A screening for non-target mutations affecting fluoroquinolone susceptibility was conducted in epidemic multidrug-resistant Salmonella enterica serovar Kentucky ST198. Among a panel of representative isolates (n = 27), covering the epidemic, only three showed distinct mutations in ramR resulting in enhanced expression of genes encoding the AcrAB-TolC efflux system and low increase in ciprofloxacin MIC. No mutations were detected in other regulatory regions of this efflux system. Ciprofloxacin...

  15. ramR mutations affecting fluoroquinolone susceptibility in epidemic multidrug-resistant Salmonella enterica serovar Kentucky ST198

    OpenAIRE

    Baucheron, Sylvie; Le Hello, Simon; Doublet, Benoît; Giraud, Etienne; Weill, François-Xavier; Cloeckaert, Axel

    2013-01-01

    International audience A screening for non-target mutations affecting fluoroquinolone susceptibility was conducted in epidemic multidrug-resistant Salmonella enterica serovar Kentucky ST198. Among a panel of representative isolates (n = 27), covering the epidemic, only three showed distinct mutations in ramR resulting in enhanced expression of genes encoding the AcrAB-TolC efflux system and low increase in ciprofloxacin MIC. No mutations were detected in other regulatory regions of this ef...

  16. ramR Mutations Affecting Fluoroquinolone Susceptibility in Epidemic Multidrug-Resistant Salmonella enterica Serovar Kentucky ST198

    OpenAIRE

    Axel eCloeckaert

    2013-01-01

    A screening for non-target mutations affecting fluoroquinolone susceptibility was conducted in epidemic multidrug-resistant Salmonella enterica serovar Kentucky ST198. Among a panel of representative isolates (n=30), covering the epidemic, only three showed distinct mutations in ramR resulting in enhanced expression of genes encoding the AcrAB-TolC efflux system and low increase in ciprofloxacin MIC. No mutations were detected in other regulatory regions of this efflux system. Ciprofloxacin r...

  17. Antimicrobial Drug Resistance of Salmonella enterica Serovar Typhi in Asia and Molecular Mechanism of Reduced Susceptibility to the Fluoroquinolones▿

    OpenAIRE

    Chau, Tran Thuy; Campbell, James Ian; Galindo, Claudia M; Van Minh Hoang, Nguyen; Diep, To Song; Nga, Tran Thu Thi; Vinh Chau, Nguyen Van; Tuan, Phung Quoc; Page, Anne Laure; Ochiai, R Leon; Schultsz, Constance; Wain, John; Bhutta, Zulfiqar A.; Parry, Christopher M.; Sujit K Bhattacharya

    2007-01-01

    This study describes the pattern and extent of drug resistance in 1,774 strains of Salmonella enterica serovar Typhi isolated across Asia between 1993 and 2005 and characterizes the molecular mechanisms underlying the reduced susceptibilities to fluoroquinolones of these strains. For 1,393 serovar Typhi strains collected in southern Vietnam, the proportion of multidrug resistance has remained high since 1993 (50% in 2004) and there was a dramatic increase in nalidixic acid resistance between ...

  18. Molecular Characterization of Extended-Spectrum Cephalosporinase-Producing Salmonella enterica Serovar Choleraesuis Isolates from Patients in Thailand and Denmark

    DEFF Research Database (Denmark)

    Sirichote, P.; Hasman, Henrik; Pulsrikarn, C.;

    2010-01-01

    The objective of this study was to characterize extended-spectrum cephalosporinase (ESC)-producing isolates of Salmonella enterica serovar Choleraesuis recovered from patients in Thailand and Denmark. Twenty-four blood culture isolates from 22 patients were included in the study, of which 23 isol...... was confirmed. Overall, a high degree of clonal diversity between isolates resistant and susceptible to cephalosporins was observed. The findings represent a serious threat to public health for the Thai people and tourists....

  19. A novel extended-spectrum TEM-type beta-lactamase, TEM-138, from Salmonella enterica serovar Infantis.

    OpenAIRE

    chouchani, chedli; Berlemont, Renaud; A. Masmoudi; Galleni, Moreno; Frère, Jean-Marie; O Belhadj; Ben-Mahrez, K.

    2006-01-01

    A novel natural TEM beta-lactamase with extended-spectrum activity, TEM-138, was identified in a ceftazidime-resistant clinical isolate of Salmonella enterica serovar Infantis. Compared to TEM-1, TEM-138 contains the following mutations: E104K, N175I, and G238S. The bla(TEM-138) gene was located on a 50-kb transferable plasmid. Expression studies with Escherichia coli revealed efficient ceftazidimase and cefotaximase activities for TEM-138.

  20. A Novel Extended-Spectrum TEM-Type β-Lactamase, TEM-138, from Salmonella enterica Serovar Infantis

    OpenAIRE

    CHOUCHANI, CHEDLY; Berlemont, Renaud; Masmoudi, Afef; Galleni, Moreno; Frere, Jean-Marie; Belhadj, Omrane; Ben-Mahrez, Kamel

    2006-01-01

    A novel natural TEM β-lactamase with extended-spectrum activity, TEM-138, was identified in a ceftazidime-resistant clinical isolate of Salmonella enterica serovar Infantis. Compared to TEM-1, TEM-138 contains the following mutations: E104K, N175I, and G238S. The blaTEM-138 gene was located on a 50-kb transferable plasmid. Expression studies with Escherichia coli revealed efficient ceftazidimase and cefotaximase activities for TEM-138.

  1. Respiratory hydrogen use by Salmonella enterica serovar Typhimurium is essential for virulence.

    Science.gov (United States)

    Maier, R J; Olczak, A; Maier, S; Soni, S; Gunn, J

    2004-11-01

    Based on available annotated gene sequence information, the enteric pathogen salmonella, like other enteric bacteria, contains three putative membrane-associated H2-using hydrogenase enzymes. These enzymes split molecular H2, releasing low-potential electrons that are used to reduce quinone or heme-containing components of the respiratory chain. Here we show that each of the three distinct membrane-associated hydrogenases of Salmonella enterica serovar Typhimurium is coupled to a respiratory pathway that uses oxygen as the terminal electron acceptor. Cells grown in a blood-based medium expressed four times the amount of hydrogenase (H2 oxidation) activity that cells grown on Luria Bertani medium did. Cells suspended in phosphate-buffered saline consumed 2 mol of H2 per mol of O2 used in the H2-O2 respiratory pathway, and the activity was inhibited by the respiration inhibitor cyanide. Molecular hydrogen levels averaging over 40 microM were measured in organs (i.e., livers and spleens) of live mice, and levels within the intestinal tract (the presumed origin of the gas) were four times greater than this. The half-saturation affinity of S. enterica serovar Typhimurium for H2 is only 2.1 microM, so it is expected that H2-utilizing hydrogenase enzymes are saturated with the reducing substrate in vivo. All three hydrogenase enzymes contribute to the virulence of the bacterium in a typhoid fever-mouse model, based on results from strains with mutations in each of the three hydrogenase genes. The introduced mutations are nonpolar, and growth of the mutant strains was like that of the parent strain. The combined removal of all three hydrogenases resulted in a strain that is avirulent and (in contrast to the parent strain) one that is unable to invade liver or spleen tissue. The introduction of one of the hydrogenase genes into the triple mutant strain on a low-copy-number plasmid resulted in a strain that was able to both oxidize H2 and cause morbidity in mice within 11

  2. Virulence and metabolic characteristics of Salmonella enterica serovar enteritidis strains with different sefD variants in hens.

    Science.gov (United States)

    Morales, Cesar A; Guard, Jean; Sanchez-Ingunza, Roxana; Shah, Devendra H; Harrison, Mark

    2012-09-01

    Salmonella enterica serovar Enteritidis is one of a few Salmonella enterica serotypes that has SEF14 fimbriae encoded by the sef operon, which consists of 4 cotranscribed genes, sefABCD, regulated by sefR. A parental strain was used to construct a sefD mutant and its complement, and all 3 strains were compared for gene expression, metabolic properties, and virulence characteristics in hens. Transcription of sefD by wild type was suppressed at 42°C and absent for the mutant under conditions where the complemented mutant had 10(3) times higher transcription. Growth of the complemented mutant was restricted in comparison to that of the mutant and wild type. Hens infected with the wild type and mutant showed decreased blood calcium and egg production, but infection with the complemented mutant did not. Thus, the absence of sefD correlated with increased metabolic capacity and enhanced virulence of the pathogen. These results suggest that any contribution that sefD makes to egg contamination is either unknown or would be limited to early transmission from the environment to the host. Absence of sefD, either through mutation or by suppression of transcription at the body temperature of the host, may contribute to the virulence of Salmonella enterica by facilitating growth on a wide range of metabolites.

  3. Survival and virulence of Salmonella enterica serovar enteritidis filaments induced by reduced water activity.

    Science.gov (United States)

    Stackhouse, Robert R; Faith, Nancy G; Kaspar, Charles W; Czuprynski, Charles J; Wong, Amy C Lee

    2012-04-01

    Salmonella enterica serovar Enteritidis strain E40 filaments were developed under conditions of a reduced water activity (a(w)) of 0.95 in tryptic soy broth (TSB) or tryptic soy agar (TSA) supplemented with 8% or 7% NaCl, respectively. Filament formation was accompanied by an increase of biomass without an increase in CFU and was affected by incubation temperature and the physical milieu. The greatest amount of filaments was recovered from TSA with 7% NaCl and incubation at 30°C. Within 2 h of transfer to fresh TSB, filaments started to septate into normal-sized cells, resulting in a rapid increase in CFU. S. Enteritidis E40 filaments were not more tolerant of low- or high-temperature stresses than nonfilamented control cells. However, there was greater survival of filaments in 10% bile salts after 24 to 48 h of incubation, during pH 2.0 acid challenge for 10 min, and under desiccation on stainless steel surfaces at 25°C and 75.5% relative humidity for 7 days. S. Enteritidis E40 filaments invaded and multiplied within Caco-2 human intestinal epithelial cells to a similar degree as control cells when a comparable CFU of filaments and control cells was used. S. Enteritidis E40 filaments established a successful infection in mice via intragastric inoculation. The filaments colonized the gastrointestinal tract and disseminated to the spleen and liver at levels comparable to those attained by control cells, even when animals were inoculated with 10- to 100-fold fewer CFU. To our knowledge this is the first demonstration of virulence of stress-induced Salmonella filaments in vitro and in vivo. Formation of filaments by Salmonella in food products and food processing environments is significant to food safety, because detection and quantitation of the pathogen may be compromised. The finding that these filaments are virulent further enhances their potential public health impact.

  4. Characterization of antimicrobial resistance in Salmonella enterica strains isolated from Brazilian poultry production.

    Science.gov (United States)

    Mattiello, Samara P; Drescher, Guilherme; Barth, Valdir C; Ferreira, Carlos A S; Oliveira, Sílvia D

    2015-11-01

    Antimicrobial resistance profiles and presence of resistance determinants and integrons were evaluated in Salmonella enterica strains from Brazilian poultry. The analysis of 203 isolates showed that those from the poultry environment (88 isolates) were significantly more resistant to antimicrobials than isolates from other sources, particularly those isolated from poultry by-product meal (106 isolates). Thirty-seven isolates were resistant to at least three antimicrobial classes. Class 1 integrons were detected in 26 isolates, and the analysis of the variable region between the 5' conserved segment (CS) and 3' CS of each class 1 integron-positive isolate showed that 13 contained a typical 3' CS and 14 contained an atypical 3' CS. One Salmonella Senftenberg isolate harbored two class 1 integrons, showing both typical and atypical 3' CSs. The highest percentage of resistance was found to sulfonamides, and sul genes were detected in the majority of the resistant isolates. Aminoglycoside resistance was detected in 50 isolates, and aadA and aadB were present in 28 and 32 isolates, respectively. In addition, strA and strB were detected in 78.1 and 65.6% isolates resistant to streptomycin, respectively. Twenty-one isolates presented reduced susceptibility to β-lactams and harbored bla(TEM), bla(CMY), and/or bla(CTX-M). Forty isolates showed reduced susceptibility to tetracycline, and most presented tet genes. These results highlight the importance of the environment as a reservoir of resistant Salmonella, which may enable the persistence of resistance determinants in the poultry production chain, contributing, therefore, to the debate regarding the impacts that antimicrobial use in animal production may exert in human health.

  5. Acute diarrhea associated with Salmonella enterica in Belo Horizonte-MG: prevalence and characterization of isolates Diarreia aguda associada a Salmonella enterica em Belo Horizonte-MG: prevalência e caracterização das amostras isoladas

    Directory of Open Access Journals (Sweden)

    Mireille Ângela Bernardes Sousa

    2013-02-01

    Full Text Available Introduction: Acute infectious diarrhea is still regarded as a public health problem associated with a wide range of etiologic agents, from which Salmonella enterica is particularly worth mentioning inasmuch as it is a major cause of inflammatory diarrhea in both developed and developing countries. Objective: To assess the distribution of S. enterica among children with acute diarrhea in Belo Horizonte and to characterize bacterium isolates. Material and methods: The study group comprised a total of 157 children from low socioeconomic background. Stool samples were collected for leukocyte analysis and Salmonella bacterial culture. The isolates were serotyped and evaluated as to antimicrobial susceptibility profile, extended-spectrum β-lactamases (ESBL production, and presence of virulence markers (invA, iroB, and spvC. RESULTS: A total of 5/3.2% children were infected by S. enterica, 3/60% by S. enterica Typhimurium, 1/20% by S. enterica Enteritidis and 1/20% S. enterica subsp. enterica serotype 8.20:z4,z23:-. Fecal leucocytes were detected in two out of five fecal specimens positive for S. enterica. Isolates from three children were resistant to nalidixic acid, nalidixic acid + chloramphenicol, and nalidixic acid + chloramphenicol + ampicillin. ESBL production was not detected. All samples presented invA and iroB genes. spvC marker was observed in isolates from two children infected by S. Typhimurium and S. Enteritidis. Conclusion: The results demonstrate that S. enterica infection is uncommon among children from our region. Furthermore, they indicate the need for periodic monitoring of bacterial antimicrobial susceptibility profile in order to establish suitable antimicrobial therapy when required.INTRODUÇÃO: A diarreia infecciosa aguda é considerada um problema de saúde pública associado a uma ampla gama de agentes etiológicos, entre os quais destaca-se Salmonella enterica, causa importante de diarreia inflamatória em pa

  6. Murein lipoprotein is a critical outer membrane component involved in Salmonella enterica serovar typhimurium systemic infection.

    Science.gov (United States)

    Fadl, A A; Sha, J; Klimpel, G R; Olano, J P; Niesel, D W; Chopra, A K

    2005-02-01

    Lipopolysaccharide (LPS) and Braun (murein) lipoprotein (Lpp) are major components of the outer membrane of gram-negative enteric bacteria that function as potent stimulators of inflammatory and immune responses. In a previous paper, we provided evidence that two functional copies of the lipoprotein gene (lppA and lppB) located on the chromosome of Salmonella enterica serovar Typhimurium contributed to bacterial virulence. In this study, we characterized lppA and lppB single-knockout (SKO) mutants and compared them with an lpp double-knockout (DKO) mutant using in vitro and in vivo models. Compared to the lpp DKO mutant, which was nonmotile, the motility of the lpp SKO mutants was significantly increased (73 to 77%), although the level of motility did not reach the level of wild-type (WT) S. enterica serovar Typhimurium. Likewise, the cytotoxicity was also significantly increased when T84 human intestinal epithelial cells and RAW264.7 murine macrophages were infected with the lpp SKO mutants compared to the cytotoxicity when cells were infected with the lpp DKO mutant. The level of interleukin-8 (IL-8) in polarized T84 cells infected with the lppB SKO mutant was significantly higher (two- to threefold higher), reaching the level in cells infected with WT S. enterica serovar Typhimurium, than the level in host cells infected with the lppA SKO mutant. The lpp DKO mutant induced minimal levels of IL-8. Similarly, sera from mice infected with the lppB SKO mutant contained 4.5- to 10-fold-higher levels of tumor necrosis factor-alpha and IL-6; the levels of these cytokines were 1.7- to 3.0-fold greater in the lppA SKO mutant-infected mice than in animals challenged with the lpp DKO mutant. The increased cytokine levels observed with the lppB SKO mutant in mice correlated with greater tissue damage in the livers and spleens of these mice than in the organs of animals infected with the lppA SKO and lpp DKO mutants. Moreover, the lppB SKO mutant-infected mice had increased

  7. A Constitutively Mannose-Sensitive Agglutinating Salmonella enterica subsp. enterica Serovar Typhimurium Strain, Carrying a Transposon in the Fimbrial Usher Gene stbC, Exhibits Multidrug Resistance and Flagellated Phenotypes

    Directory of Open Access Journals (Sweden)

    Kuan-Hsun Wu

    2012-01-01

    Full Text Available Static broth culture favors Salmonella enterica subsp. enterica serovar Typhimurium to produce type 1 fimbriae, while solid agar inhibits its expression. A transposon inserted in stbC, which would encode an usher for Stb fimbriae of a non-flagellar Salmonella enterica subsp. enterica serovar Typhimurium LB5010 strain, conferred it to agglutinate yeast cells on both cultures. RT-PCR revealed that the expression of the fimbrial subunit gene fimA, and fimZ, a regulatory gene of fimA, were both increased in the stbC mutant when grown on LB agar; fimW, a repressor gene of fimA, exhibited lower expression. Flagella were observed in the stbC mutant and this phenotype was correlated with the motile phenotype. Microarray data and RT-PCR indicated that the expression of three genes, motA, motB, and cheM, was enhanced in the stbC mutant. The stbC mutant was resistant to several antibiotics, consistent with the finding that expression of yhcQ and ramA was enhanced. A complementation test revealed that transforming a recombinant plasmid possessing the stbC restored the mannose-sensitive agglutination phenotype to the stbC mutant much as that in the parental Salmonella enterica subsp. enterica serovar Typhimurium LB5010 strain, indicating the possibility of an interplay of different fimbrial systems in coordinating their expression.

  8. A murine model to study the antibacterial effect of copper on infectivity of Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Sharan, Riti; Chhibber, Sanjay; Reed, Robert H

    2011-01-01

    This study investigated the effect of copper as an antibacterial agent on the infectivity of Salmonella enterica serovar Typhimurium. Mice were infected orally with a standardized dose of unstressed Salmonella Typhimurium and copper-stressed cells of Salmonella Typhimurium. Bacterial counts in ileum, blood, liver and spleen were observed up to 168 h under normal aerobic conditions. Serum sensitivity, phagocytosis, malondialdehyde levels and histopathology were studied for both set of animals. A decreased bacterial count in the organs with mild symptoms of infection and a complete recovery by 48 h was observed in mice infected with copper-stressed bacteria. Histopathological examination of ileum tissue demonstrated regeneration of damaged tissue post-infection with copper-stressed bacteria and no malondialdehyde levels were detected after 24 h in ileum, spleen and liver. Exposure to copper sensitized Salmonella Typhimurium to the lytic action of serum and intracellular killing by peritoneal macrophages. It can be concluded that copper stress confers a decrease in the infectivity of healthy Salmonella Typhimurium in normal mice. This study highlights the significance of use of copper as an antibacterial agent against Salmonella Typhimurium in reducing the risk of incidence of Salmonella infections from contaminated water.

  9. A Murine Model to Study the Antibacterial Effect of Copper on Infectivity of Salmonella Enterica Serovar Typhimurium

    Directory of Open Access Journals (Sweden)

    Robert H. Reed

    2010-12-01

    Full Text Available This study investigated the effect of copper as an antibacterial agent on the infectivity of Salmonella enterica serovar Typhimurium. Mice were infected orally with a standardized dose of unstressed Salmonella Typhimurium and copper-stressed cells of Salmonella Typhimurium. Bacterial counts in ileum, blood, liver and spleen were observed up to 168 h under normal aerobic conditions. Serum sensitivity, phagocytosis, malondialdehyde levels and histopathology were studied for both set of animals. A decreased bacterial count in the organs with mild symptoms of infection and a complete recovery by 48 h was observed in mice infected with copper-stressed bacteria. Histopathological examination of ileum tissue demonstrated regeneration of damaged tissue post-infection with copper-stressed bacteria and no malondialdehyde levels were detected after 24 h in ileum, spleen and liver. Exposure to copper sensitized Salmonella Typhimurium to the lytic action of serum and intracellular killing by peritoneal macrophages. It can be concluded that copper stress confers a decrease in the infectivity of healthy Salmonella Typhimurium in normal mice. This study highlights the significance of use of copper as an antibacterial agent against Salmonella Typhimurium in reducing the risk of incidence of Salmonella infections from contaminated water.

  10. A comparison of transmission characteristics of Salmonella enterica serovar Enteritidis between pair-housed and group-housed laying hens

    Directory of Open Access Journals (Sweden)

    Thomas Ekelijn

    2011-02-01

    Full Text Available Abstract Human cases of bacterial gastro-enteritis are often caused by the consumption of eggs contaminated with Salmonella species, mainly Salmonella enterica serovar Enteriditis (Salmonella Enteritidis. To reduce human exposure, in several countries worldwide surveillance programmes are implemented to detect colonized layer flocks. The sampling schemes are based on the within-flock prevalence, and, as this changes over time, knowledge of the within-flock dynamics of Salmonella Enteritidis is required. Transmission of Salmonella Enteritidis has been quantified in pairs of layers, but the question is whether the dynamics in pairs is comparable to transmission in large groups, which are more representative for commercial layer flocks. The aim of this study was to compare results of transmission experiments between pairs and groups of laying hens. Experimental groups of either 2 or 200 hens were housed at similar densities, and 1 or 4 hens were inoculated with Salmonella Enteritidis, respectively. Excretion was monitored by regularly testing of fecal samples for the presence of Salmonella Enteritidis. Using mathematical modeling, the group experiments were simulated with transmission parameter estimates from the pairwise experiments. Transmission of the bacteria did not differ significantly between pairs or groups. This finding suggests that the transmission parameter estimates from small-scale experiments might be extrapolated to the field situation.

  11. A comparison of transmission characteristics of Salmonella enterica serovar Enteritidis between pair-housed and group-housed laying hens.

    Science.gov (United States)

    Thomas, Ekelijn; Bouma, Annemarie; Klinkenberg, Don

    2011-02-23

    Human cases of bacterial gastro-enteritis are often caused by the consumption of eggs contaminated with Salmonella species, mainly Salmonella enterica serovar Enteriditis (Salmonella Enteritidis). To reduce human exposure, in several countries worldwide surveillance programmes are implemented to detect colonized layer flocks. The sampling schemes are based on the within-flock prevalence, and, as this changes over time, knowledge of the within-flock dynamics of Salmonella Enteritidis is required. Transmission of Salmonella Enteritidis has been quantified in pairs of layers, but the question is whether the dynamics in pairs is comparable to transmission in large groups, which are more representative for commercial layer flocks. The aim of this study was to compare results of transmission experiments between pairs and groups of laying hens. Experimental groups of either 2 or 200 hens were housed at similar densities, and 1 or 4 hens were inoculated with Salmonella Enteritidis, respectively. Excretion was monitored by regularly testing of fecal samples for the presence of Salmonella Enteritidis. Using mathematical modeling, the group experiments were simulated with transmission parameter estimates from the pairwise experiments. Transmission of the bacteria did not differ significantly between pairs or groups. This finding suggests that the transmission parameter estimates from small-scale experiments might be extrapolated to the field situation.

  12. Reduction of Salmonella enterica serovar enteritidis on the surface of raw shelled almonds by exposure to steam.

    Science.gov (United States)

    Lee, Sun-Young; Oh, Se-wook; Chung, Hyun-Jung; Reyes-De-Corcuera, Jose I; Powers, Joseph R; Kang, Dong-Hyun

    2006-03-01

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

  13. Antibiotic resistances in Listeria monocytogenes and Salmonella enterica isolated from foods with animal origin Resistencias a antibióticos en Listeria monocytogenes y Salmonella enterica aislados de alimentos de origen animal

    Directory of Open Access Journals (Sweden)

    Baltasar Balsalobre Hernández

    2004-12-01

    Full Text Available Extensive use of antibiotics in both human and animal health and in cattle production has generated resistant microorganisms to common antibiotics. Resistances spread caused by human and animal therapeutic is well known, but we know poorly frecuency of resistant bacteria in foods with animal origin and destinated to human consumers. In this paper, sensitivity to nineteen antibiotics was investigated in Listeria monocytogenes and Salmonella enterica strains isolated from foods with animal origin, including fresh meat, hamburgers, fresh sausages, boiled ham and new-laid chicken eggs. The plate diffusion method of Bauer-Kirby was used.Listeria monocytogenes strains showed a very high sensitivity to all antibiotics checked, with the exception of one strain tetracycline resistant. In contrast, Salmonella enterica showed a high frecuency of resistances, in special to tetracycline, streptomycin, nalidixic acid, ticarcillin, ampicillin and chloramphenicol. Moreover, multi-resistance was a common phenomenon. Twenty percent of S. enterica strains were resistant to four or more antibiotics. Frecuency of resistances was higher in 4,5,12:i:-, Hadar, Typhimurium and Virchow serotypes.In conclusion, Salmonella enterica strains isolated from foods with animal origin and destinated to human consumers are usually resistant to several antibiotics. The significance of this observation and its potential health risk must be investigated.El uso extensivo de antibióticos para la salud humana y animal así como para mejorar la producción ganadera ha generado un gran número de cepas microbianas resistentes a antibióticos de uso común. Es bien conocida la difusión de resistencias a través de la terapéutica humana y animal, pero desconocemos en qué medida los alimentos de origen animal destinados a consumo humano son portadores de resistencias.En este trabajo, se investigó la sensibilidad a diecinueve antibióticos de cepas de Listeria monocytogenes y Salmonella

  14. Salmonella enterica serovar Typhimurium and Escherichia coli contamination of root and leaf vegetables grown in soils with incorporated bovine manure.

    Science.gov (United States)

    Natvig, Erin E; Ingham, Steven C; Ingham, Barbara H; Cooperband, Leslie R; Roper, Teryl R

    2002-06-01

    Bovine manure, with or without added Salmonella enterica serovar Typhimurium (three strains), was incorporated into silty clay loam (SCL) and loamy sand (LS) soil beds (53- by 114-cm surface area, 17.5 cm deep) and maintained in two controlled-environment chambers. The S. enterica serovar Typhimurium inoculum was 4 to 5 log CFU/g in manure-fertilized soil. The conditions in the two environmental chambers, each containing inoculated and uninoculated beds of manure-fertilized soil, simulated daily average Madison, Wis., weather conditions (hourly temperatures, rainfall, daylight, and humidity) for a 1 March or a 1 June manure application and subsequent vegetable growing seasons ending 9 August or 28 September, respectively. Core soil samples were taken biweekly from both inoculated and uninoculated soil beds in each chamber. Radishes, arugula, and carrots were planted in soil beds, thinned, and harvested. Soils, thinned vegetables, and harvested vegetables were analyzed for S. enterica serovar Typhimurium and Escherichia coli (indigenous in manure). After the 1 March manure application, S. enterica serovar Typhimurium was detected at low levels in both soils on 31 May, but not on vegetables planted 1 May and harvested 12 July from either soil. After the 1 June manure application, S. enterica serovar Typhimurium was detected in SCL soil on 7 September and on radishes and arugula planted in SCL soil on 15 August and harvested on 27 September. In LS soil, S. enterica serovar Typhimurium died at a similar rate (P >or= 0.05) after the 1 June manure application and was less often detected on arugula and radishes harvested from this soil compared to the SCL soil. Pathogen levels on vegetables were decreased by washing. Manure application in cool (daily average maximum temperature of 20 degrees C) summer conditions is not recommended when vegetable planting is done between the time of manure application and late summer. A late fall manure application will not increase the

  15. Detection of Salmonella enterica in Magellanic penguins (Spheniscus magellanicus) of Chilean Patagonia: evidences of inter-species transmission.

    Science.gov (United States)

    Dougnac, C; Pardo, C; Meza, K; Arredondo, C; Blank, O; Abalos, P; Vidal, R; Fernandez, A; Fredes, F; Retamal, P

    2015-04-01

    Patagonia in southern South America is among the few world regions where direct human impact is still limited but progressively increasing, mainly represented by tourism, farming, fishing and mining activities. The sanitary condition of Patagonian wildlife is unknown, in spite of being critical for the assessment of anthropogenic effects there. The aim of this study was the characterization of Salmonella enterica strains isolated from wild colonies of Magellanic penguins (Spheniscus magellanicus) located in Magdalena Island and Otway Sound, in Chilean Patagonia. Eight isolates of Salmonella were found, belonging to Agona and Enteritidis serotypes, with an infection rate of 0·38%. Resistance to ampicillin, cefotaxime, ceftiofur and tetracycline antimicrobials were detected, and some of these strains showed genotypic similarity with Salmonella strains isolated from humans and gulls, suggesting inter-species transmission cycles and strengthening the role of penguins as sanitary sentinels in the Patagonian ecosystem.

  16. Detection of Salmonella enterica in Magellanic penguins (Spheniscus magellanicus) of Chilean Patagonia: evidences of inter-species transmission.

    Science.gov (United States)

    Dougnac, C; Pardo, C; Meza, K; Arredondo, C; Blank, O; Abalos, P; Vidal, R; Fernandez, A; Fredes, F; Retamal, P

    2015-04-01

    Patagonia in southern South America is among the few world regions where direct human impact is still limited but progressively increasing, mainly represented by tourism, farming, fishing and mining activities. The sanitary condition of Patagonian wildlife is unknown, in spite of being critical for the assessment of anthropogenic effects there. The aim of this study was the characterization of Salmonella enterica strains isolated from wild colonies of Magellanic penguins (Spheniscus magellanicus) located in Magdalena Island and Otway Sound, in Chilean Patagonia. Eight isolates of Salmonella were found, belonging to Agona and Enteritidis serotypes, with an infection rate of 0·38%. Resistance to ampicillin, cefotaxime, ceftiofur and tetracycline antimicrobials were detected, and some of these strains showed genotypic similarity with Salmonella strains isolated from humans and gulls, suggesting inter-species transmission cycles and strengthening the role of penguins as sanitary sentinels in the Patagonian ecosystem. PMID:25148565

  17. Reduction of Salmonella enterica on the surface of eggshells by sequential treatment with aqueous chlorine dioxide and drying.

    Science.gov (United States)

    Choi, Seonyeong; Park, Sunhyung; Kim, Yoonsook; Kim, Byeong-sam; Beuchat, Larry R; Hoikyung, Kim; Ryu, Jee-Hoon

    2015-10-01

    The synergistic effects of sequential treatments with chlorine dioxide (ClO2) and drying in killing Salmonella enterica on the surface of chicken eggshells were investigated. Initial experiments were focused on comparing lethalities of sodium hypochlorite (NaOCl) and ClO2. Eggs surface-inoculated with S. enterica in chicken feces as a carrier were immersed in water, NaOCl (50 or 200 μg/mL), or ClO2 (50 or 200 μg/mL) for 1 or 5 min. For 1-min treatments, lethal activities of sanitizers were not significantly different (P>0.05). However, after treatment with ClO2 for 5 min, reductions of S. enterica were significantly greater (P≤0.05) than reductions after treatment with water or NaOCl. The effect of treatment of eggs with ClO2 or NaOCl, followed by drying at 43% relative humidity and 25 °C for 24 and 48 h, were determined. Populations of S. enterica decreased during drying, regardless of the type of sanitizer treatment. ClO2 treatment, compared to water or NaOCl treatments, resulted in additional reductions of ca. >1.3 log CFU/egg during drying. This indicates that sequential treatments with ClO2 and drying induced synergistic lethal effects against S. enterica on the surface of eggshells. These observations will be useful when selecting a sanitizer to control S. enterica on the surface of eggshells and designing an effective egg sanitization system exploiting the synergistic lethal effects of sanitizer and drying. PMID:26114591

  18. Antibacterial activity of honey against clinical isolates of Escherichia coli, Pseudomonas aeruginosa and Salmonella enterica serovar Typhi

    Institute of Scientific and Technical Information of China (English)

    Shyamapada Mandal; Manisha DebMandal; Nishith Kumar Pal; Krishnendu Saha

    2010-01-01

    Objective: To ascertain the potential antibacterial activity of honey against clinical isolates ofEscherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa) and Salmonella enterica serovar Typhi (S. enterica serovar Typhi) by in vitro methods. Methods: The partial inhibitory concentration (PIC), minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of the autoclaved honey (extracted from Apis indica hive by indigenous method) were determined for S. enterica serovar Typhi (n=8; from blood cultute),E. coli (n=5; from urine culture) and P. aeruginosa (n=5; from pus culture) isolates by in vitro methods. Results: The PICs of the honey tested for the isolates ranged 0.50%-1.25 % (v/v) for S. enterica serovar Typhi, 0.75%-1.50% (v/v) for E. coli and 1.00%-1.25 % (v/v) for P. aeruginosa, while the MICs ranged 1.75%-3.00% (v/v), 3.00%-3.50% (v/v) and 3.50% (v/v), respectively. The P. aeruginosa and E. coli isolates had MBC value of 4.00% (v/v); the S. enterica serovar Typhi showed MBCs in between 3.00% and 3.50% (v/v). The bactericidal activity of honey was achieved at concentration 3.00% (v/v) for S. enterica serovar Typhi and E. coli, and at 3.50% (v/v) for P. aeruginosa. Conclusions: The excellent antibacterial activity of honey against clinical bacterial isolates indicates the usefulness of honey in clinical practice against bacterial infection.

  19. High-throughput molecular determination of salmonella enterica serovars by use of multiplex PCR and capillary electrophoresis analysis.

    Science.gov (United States)

    Leader, Brandon T; Frye, Jonathan G; Hu, Jinxin; Fedorka-Cray, Paula J; Boyle, David S

    2009-05-01

    Salmonella enterica is a leading cause of food-borne illness worldwide and is also a major cause of morbidity and mortality in domestic and wild animals. In the current study, a high-throughput molecular assay was developed to determine the most common clinical and nonhuman serovars of S. enterica in the United States. Sixteen genomic targets were identified based on their differential distribution among common serovars. Primers were designed to amplify regions of each of these targets in a single multiplex PCR while incorporating a 6-carboxyfluorescein-labeled universal primer to fluorescently label all amplicons. The fluorescently labeled PCR products were separated using capillary electrophoresis, and a Salmonella multiplex assay for rapid typing (SMART) code was generated for each isolate, based upon the presence or absence of PCR products generated from each target gene. Seven hundred fifty-one blind clinical isolates of Salmonella from Washington State, collected in 2007 and previously serotyped via antisera, were screened with the assay. A total of 89.6% of the isolates were correctly identified based on comparison to a panel of representative SMART codes previously determined for the top 50 most common serovars in the United States. Of the remaining isolates, 6.2% represented isolates that produced a new SMART code for a previously determined serotype, while the final 8.8% were from serotypes not screened in the original panel used to score isolates in the blinded study. This high-throughput multiplex PCR assay allowed simple and accurate typing of the most prevalent clinical serovars of Salmonella enterica at a level comparable to that of conventional serotyping, but at a fraction of both the cost and time required per test. PMID:19261787

  20. Role in virulence and protective efficacy in pigs of Salmonella enterica serovar Typhimurium secreted components identified by signature-tagged mutagenesis.

    Science.gov (United States)

    Carnell, Sonya C; Bowen, Alison; Morgan, Eirwen; Maskell, Duncan J; Wallis, Timothy S; Stevens, Mark P

    2007-06-01

    Salmonella enterica serovar Typhimurium (S. Typhimurium) is a zoonotic enteric pathogen of worldwide importance and pigs are a significant reservoir of human infection. Signature-tagged transposon mutagenesis (STM) was used to identify genes required by S. Typhimurium to colonize porcine intestines. A library of 1045 signature-tagged mutants of S. Typhimurium ST4/74 Nal(R) was screened following oral inoculation of pigs in duplicate. A total of 119 attenuating mutations were identified in 95 different genes, many of which encode known or putative secreted or surface-anchored molecules. A large number of attenuating mutations were located within Salmonella pathogenicity islands (SPI)-1 and -2, confirming important roles for type III secretion systems (T3SS)-1 and -2 in intestinal colonization of pigs. Roles for genes encoded in other pathogenicity islands and islets, including the SPI-6-encoded Saf atypical fimbriae, were also identified. Given the role of secreted factors and the protection conferred against other pathogens by vaccination with extracellular and type III secreted proteins, the efficacy of a secreted protein vaccine from wild-type S. Typhimurium following intramuscular vaccination of pigs was evaluated. Serum IgG responses against type III secreted proteins were induced following vaccination and a significant reduction in faecal excretion of S. Typhimurium was observed in the acute phase of infection compared to mock-vaccinated animals. Vaccination with secreted proteins from an isogenic S. Typhimurium prgH mutant produced comparable levels of protection to vaccination with the preparation from the parent strain, indicating that protection was not reliant on T3SS-1 secreted proteins. The data provide valuable information for the control of Salmonella in pigs.

  1. Iranian clonal population of Salmonella enterica serovar Enteritidis, char- acterized by multi-locus sequence typing (MLST method

    Directory of Open Access Journals (Sweden)

    Rainak Ghaderi

    2015-12-01

    Full Text Available Background and Objectives: Some 2 million tons of chicken meat is produced by Iran per annum, positioning Iran among the top producers in the region. This study aimed to evaluate the molecular epidemiology and genetic characteristics of Sal- monella enterica Enteritidis in Iran.Materials and Methods: A representative selection of isolates (n=76, initially genotyped by a 7-locus MLVA typing sys- tem, was examined by the standard MLST genotyping.Results and Conclusion: All the MLVA typed isolates, classified into six types, were gathered under a single ST11 MLST type. This is an intriguing observation as much more genome heterogeneity was expected considering the extent of diversity in the host and geography origin of the examined isolates. ST11, on the other hand is not exclusively found in Iran as it is reported also from Brazil, Denmark, Japan and the United States. In explanation of these observations, ST11 might stand for a single probably ancestral clone of Salmonella enterica Enteritidis successfully scattered in all these geographically diverse countries. Further global investigation covering more isolates and methods like whole genome sequencing would be advisable. Keywords: Salmonella enterica Enteritidis, MLST, MLVA, Genetic diversity

  2. Differences in the early response of hatchlings of different chicken breeding lines to Salmonella enterica serovar Enteritidis infection.

    Science.gov (United States)

    Schokker, D; Peters, T H F; Hoekman, A J W; Rebel, J M J; Smits, M A

    2012-02-01

    Poultry products are the major source of food-borne Salmonella infection in humans. Broiler lines selected to be more resistant to Salmonella could reduce the transfer of Salmonella to humans. To investigate differences in the susceptibility of newly hatched chicks to oral infection with Salmonella enterica serovar Enteritidis, 3 commercial broiler lines (A, B, and C) were infected immediately after hatch and compared to healthy controls at 0.33, 1, and 2 d postinfection. Weight, bacteriological examination, and the jejunal influx of CD4, CD8, TCRαβ, TCRγδ, and KUL01 (macrophages and dendritic cells) cells that are positive was investigated. In addition, the jejunal transcriptional response was analyzed using whole-genome chicken cDNA arrays. Salmonella colony-forming unit counts from cecal content and liver revealed that Salmonella enterica entered the body at 0.33 d postinfection. Broiler line A appeared most susceptible to intestinal colonization and the systemic spread of Salmonella. In addition, the Salmonella-induced jejunal influx of macrophages in this line showed a clear increase in time, which is in contrast to lines B and C. On the other hand, all lines showed a peak of CD4(+) cells at 1 d postinfection when infected chicks were compared to control chicks. The transcriptional response of line A clearly differed from the responses in lines B and C. Functional analysis indicated that the majority of the differentially expressed genes at 0.33 d postinfection in line A were involved in cell-cycle functions, whereas at 2 d postinfection the majority of the differentially expressed genes could be assigned to inflammatory disorder, differentiation and proliferation of (T) lymphocytes. These data indicate that hatchlings of different broiler lines differ in their systemic spread of Salmonella and suggest that intestinal barrier functions, as well as immunological responses, may be the underlying factors. We hypothesize that the differences between genetic

  3. Isolation of Salmonella enterica and serologic reactivity to Leptospira interrogans in opossums (Didelphis virginiana from Yucatán, México Aislamiento de Salmonella enterica y reactividad serológica a Leptospira interrogans en tlacuaches (Didelphis virginiana de Yucatán, México

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    Hugo Antonio RUIZ-PIÑA

    2002-07-01

    Full Text Available The presence of Salmonella enterica and serologic evidence of infection by Leptospira interrogans, were detected in the opossum Didelphis virginiana in a semi-urban locality of the Yucatán State, México. Ninety-one opossums were captured during the period April 1996 and May 1998. From a total of 17 feces samples, four Salmonella enterica subsp. enterica serotypes (Sandiego, Newport, Anatum, and Minnesota, and one Salmonella enterica subsp. arizonae serovar O44:Z4,Z23:- were isolated. Some opossums presented mixed infections. From 81 sera samples, four (4.9% were positive to antibodies to Leptospira serovars pomona and wolfii. Both animals infected with Salmonella enterica and those serologically positive to Leptospira interrogans were captured in peridomestic habitat. Opossums infected with Salmonella enterica, were captured in dry season, and those seropositive to Leptospira interrogans during the rainy season. The implications of infection and reactivity of these zoonotic pathogens in D. virginiana in the Yucatan state are briefly discussed.La presencia de Salmonella enterica y evidencia serológica de infección por Leptospira interrogans fueron detectadas en tlacuaches de la especie Didelphis virginiana capturados en una localidad semi-urbana del estado de Yucatán, México. Se capturaron 91 marsupiales durante el período de abril de 1996 a mayo de 1998. De un total de 17 muestras de heces, se aislaron cuatro serotipos de Salmonella enterica subsp. enterica (Sandiego, Newport, Anatum y Minnesota y una Salmonella enterica subsp. arizonae serovar O44:Z4,Z23:-. En algunos tlacuaches se registraron infecciones mixtas. De 81 muestras de suero, cuatro (4,9% presentaron reacciones positivas con los serovares pomona y wolffi, ambos pertenecientes al género Leptospira. Los tlacuaches con serología positiva fueron capturados en el hábitat peridomiciliar. Los animales infectados con Salmonella enterica fueron capturados en los períodos de seca y

  4. Salmonella enterica serovar Enteritidis enterocolitis during late stages of gestation induces an adverse pregnancy outcome in the murine model.

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    Mariángeles Noto Llana

    Full Text Available Foodborne diseases caused by Salmonella enterica serovar Enteritidis (S. Enteritidis are a significant health problem. Pregnancy, state of immunological tolerance, is a predisposing condition for the development of infections with intracellular pathogens. Salmonella species can cause pregnancy complications such as chorioamnionitis, transplacental fetal infection, pre term labor, abortions, neonatal and maternal septicemia. However, the specific mechanisms by which Salmonella infections trigger these alterations are not clear. In the present work, using a self-limiting enterocolitis murine model, we show that the ingestion of a low dose of S. Enteritidis at late stages of pregnancy (day 15 of gestation is sufficient to induce massive maternal infection. We found that Salmonella infection leads to 40% of pre term delivery, 33% of abortion and fetal growth restriction. Placental dysfunction during S. Enteritidis enterocolitis was confirmed through cellular infiltration and hypoxia markers (MPO activity and COX-1 and COX-2 expression, respectively. Apoptosis in placental tissue due to Salmonella infection was also evident at day 18 of gestation when investigated by morphometric procedure, DNA fragmentation and Fas/FasL expression. Also, the expression of IFN-γ, TNF-α, IL-17 and IL-10 was up regulated in response to Salmonella not only in placenta, but also in amniotic fluid and maternal serum. Altogether, our results demonstrate that S. Enteritidis enterocolitis during late stages of gestation causes detrimental effect on pregnancy outcome.

  5. Molecular characteristics of Salmonella enterica Paratyphi A in Yunnan Province, southwest China.

    Science.gov (United States)

    Gu, Wenpeng; Yang, Zushun; Chen, Yujuan; Yin, Jianwen; Yang, Jianbin; Li, Chaoqun; Zhou, Yongming; Yin, Jie; Xu, Wen; Zhao, Shiwen; Liang, Junrong; Wang, Xin; Jing, Huaiqi; Fu, Xiaoqing

    2015-03-01

    Previously, the prevalence of Salmonella enterica Paratyphi A in Yunnan was high; and recently Yunnan was the predominant endemic province in China. To identify the molecular epidemiology, antibiotic resistance profile and genotypic diversity of the S. Paratyphi A isolates from 1995 to 2013 in Yunnan, we performed the study. Antibiotic susceptibility tests, pulse-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to identify the characteristics of the bacterial isolates. The results showed from 1995 to 2013, 366 S. Paratyphi A were isolated: 295 isolates (80.6%) from Yuxi and 68 isolates (18.58%) from Honghe. All of the strains were resistant to nalidixic acid, and some were resistant to ampicillin and trimethoprim/sulfamethoxazole in different years. All the isolates were sensitive to cefotaxime and ciprofloxacin. Identical PFGE with two enzyme digestion patterns were found for 339 isolates. Some environmental isolates in different years were homologous with the strains isolated from food and patients. MLST showed 349 strains were ST85, only 17 isolates were ST129. S. Paratyphi A isolates from Yunnan showed a high similarity, and we found the pathogen isolated from patients, the environment and food had the close epidemiological relationship, forming a transmission circulation. These findings have important implications for paratyphoid-control strategies.

  6. Lack of efflux mediated quinolone resistance in Salmonella enterica serovars Typhi and Paratyphi A

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    Sylvie eBaucheron

    2014-01-01

    Full Text Available Salmonella enterica serovars Typhi and Paratyphi A isolates from human patients in France displaying different levels of resistance to quinolones or fluoroquinolones were studied for resistance mechanisms to these antimicrobial agents. All resistant isolates carried either single or multiple target gene mutations (i.e. in gyrA, gyrB, or parC correlating with the resistance levels observed. Active efflux, through upregulation of multipartite efflux systems, has also been previously reported as contributing mechanism for other serovars. Therefore, we investigated also the occurrence of non-target gene mutations in regulatory regions affecting efflux pump expression. However, no mutation was detected in these regions in both Typhi and Paratyphi isolates of this study. Besides, no overexpression of the major efflux systems was observed for these isolates. Nevertheless, a large deletion of 2334 bp was identified in the acrS-acrE region of all S. Typhi strains but which did not affect the resistance phenotype. As being specific to S. Typhi, this deletion could be used for specific molecular detection purposes. In conclusion, the different levels of quinolone or FQ resistance in both S. Typhi and S. Paratyphi A seem to rely only on target modifications.

  7. Influence of Temperature, Source, and Serotype on Biofilm Formation of Salmonella enterica Isolates from Pig Slaughterhouses.

    Science.gov (United States)

    Piras, Francesca; Fois, Federica; Consolati, Simonetta Gianna; Mazza, Roberta; Mazzette, Rina

    2015-10-01

    Quantitative assessment of in vitro biofilm formation by 40 Salmonella enterica isolates isolated in pig abattoirs from animal and environmental sources (surfaces in contact and not in contact with meat) and classified in eight seroytpes was carried out by using a microtiter plate assay with spectrophotometric reading (optical density at 620 nm). Biofilm-forming ability was statistically correlated with the temperature of incubation (22 and 35°C), the source of the isolates, and the antimicrobial resistance profile. After incubation at 35°C, 9 isolates (22.5%) were classified as weak biofilm producers. After incubation at 22°C, 25 isolates (62.5%) were classified as weak producers and 3 (7.5%) as moderate producers. The quantity of biofilm formed after incubation at 22°C was significantly higher (P < 0.01) than at 35°C. This result is notable because 22°C is a common temperature in meat processing facilities and in slaughterhouses. At 35°C, isolates detected from surfaces in contact with meat showed significantly higher (P < 0.1) optical density values compared to isolates from other samples, highlighting the risk of cross-contamination for carcasses and offal. No correlation was detected between quantity of biofilm and serotype or between biofilm formation and resistance to antimicrobials.

  8. Genomic variants of bacteriophages against Salmonella enterica serovar Enteritidis with potential application in the poultry industry

    Directory of Open Access Journals (Sweden)

    J Robeson

    2008-09-01

    Full Text Available Salmonella enterica serovar Enteritidis (SE is a prevalent gastrointestinal pathogen worldwide, threatening both animal and human health. In the latter, disease is associated to the consumption of SE-contaminated products from the poultry industry. The control of SE infection is largely based on the use of antibiotics and vaccines, but the use of lytic bacteriophages is re-emerging as an additional strategy for SE control. In fact, a number of recent reports point to the adequacy of bacteriophage as an efficient prophylactic or therapeutic countermeasure to SE infections. However, less attention has been focused on the basic biology of these bacteriophages. Here we report on three bacteriophages (f18, IF1 and EST2 that share a common viral particle morphology but are genomic variants as judged by their EcoRI DNA restriction patterns. Furthermore, they differ in their lytic capability towards SE, being EST2 the most efficient. They show a very narrow host range, efficiently infecting only SE strains. In terms of stability in various suspension media, including distilled water, all three bacteriophages remained viable, without noticeable decay in titer for at least 15 days at 25ºC. These results suggest the suitability of the tested bacteriophages as SE-controlling agents in the poultry industry.

  9. Inactivation of Salmonella enterica serovar Typhimurium on fresh produce by cold atmospheric gas plasma technology.

    Science.gov (United States)

    Fernández, A; Noriega, E; Thompson, A

    2013-02-01

    Cold atmospheric gas plasma treatment (CAP) is an alternative approach for the decontamination of fresh and minimally processed food. In this study, the effects of growth phase, growth temperature and chemical treatment regime on the inactivation of Salmonella enterica serovar Typhimurium (S. Typhimurium) by Nitrogen CAP were examined. Furthermore, the efficacy of CAP treatment for decontaminating lettuce and strawberry surfaces and potato tissue inoculated with S. Typhimurium was evaluated. It was found that the rate of inactivation of S. Typhimurium was independent of the growth phase, growth temperature and chemical treatment regime. Under optimal conditions, a 2 min treatment resulted in a 2.71 log-reduction of S. Typhimurium viability on membrane filters whereas a 15 min treatment was necessary to achieve 2.72, 1.76 and 0.94 log-reductions of viability on lettuce, strawberry and potato, respectively. We suggest that the differing efficiency of CAP treatment on the inactivation of S. Typhimurium on these different types of fresh foods is a consequence of their surface features. Scanning electron microscopy of the surface structures of contaminated samples of lettuce, strawberry and potato revealed topographical features whereby S. Typhimurium cells could be protected from the active species generated by plasma.

  10. Solvent accessibility and purifying selection within proteins of Escherichia coli and Salmonella enterica.

    Science.gov (United States)

    Bustamante, C D; Townsend, J P; Hartl, D L

    2000-02-01

    The neutral theory of molecular evolution predicts that variation within species is inversely related to the strength of purifying selection, but the strength of purifying selection itself must be related to physical constraints imposed by protein folding and function. In this paper, we analyzed five enzymes for which polymorphic sequence variation within Escherichia coli and/or Salmonella enterica was available, along with a protein structure. Single and multivariate logistic regression models are presented that evaluate amino acid size, physicochemical properties, solvent accessibility, and secondary structure as predictors of polymorphism. A model that contains a positive coefficient of association between polymorphism and solvent accessibility and separate intercepts for each secondary-structure element is sufficient to explain the observed variation in polymorphism between sites. The model predicts an increase in the probability of amino acid polymorphism with increasing solvent accessibility for each protein regardless of physicochemical properties, secondary-structure element, or size of the amino acid. This result, when compared with the distribution of synonymous polymorphism, which shows no association with solvent accessibility, suggests a strong decrease in purifying selection with increasing solvent accessibility. PMID:10677853

  11. Identification of in vivo induced protein antigens of Salmonella enterica serovar Typhi during human infection

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    During infectious disease episodes, pathogens express distinct subsets of virulence factors which allow them to adapt to different environments. Hence, genes that are expressed or upregulated in vivo are implicated in pathogenesis. We used in vivo induced antigen technology (IVIAT) to identify antigens which are expressed during infection with Salmonella enterica serovar Typhi. We identified 7 in vivo induced (IVI) antigens, which included BcfD (a fimbrial structural subunit), GrxC (a glutaredoxin 3), SapB (an ABC-type transport system), T3663 (an ABC-type uncharacterized transport system), T3816 (a putative rhodanese-related sulfurtransferase), T1497 (a probable TonB-dependent receptor) and T3689 (unknown function). Of the 7 identified antigens, 5 antigens had no cross-immunoreactivity in adsorbed control sera from healthy subjects. These 5 included BcfD, GrxC, SapB, T3663 and T3689. Antigens identified in this study are potential targets for drug and vaccine development and may be utilized as diagnostic agents.

  12. Biofilm formation ability of Salmonella enterica serovar Typhimurium acrAB mutants.

    Science.gov (United States)

    Schlisselberg, Dov B; Kler, Edna; Kisluk, Guy; Shachar, Dina; Yaron, Sima

    2015-10-01

    Recent studies offer contradictory findings about the role of multidrug efflux pumps in bacterial biofilm development. Thus, the aim of this study was to investigate the involvement of the AcrAB efflux pump in biofilm formation by investigating the ability of AcrB and AcrAB null mutants of Salmonella enterica serovar Typhimurium to produce biofilms. Three models were used to compare the ability of S. Typhimurium wild-type and its mutants to form biofilms: formation of biofilm on polystyrene surfaces; production of biofilm (mat model) on the air/liquid interface; and expression of curli and cellulose on Congo red-supplemented agar plates. All three investigated genotypes formed biofilms with similar characteristics. However, upon exposure to chloramphenicol, formation of biofilms on solid surfaces as well as the production of curli were either reduced or were delayed more significantly in both mutants, whilst there was no visible effect on pellicle formation. It can be concluded that when no selective pressure is applied, S. Typhimurium is able to produce biofilms even when the AcrAB efflux pumps are inactivated, implying that the use of efflux pump inhibitors to prevent biofilm formation is not a general solution and that combined treatments might be more efficient. Other factors that affect the ability to produce biofilms depending on efflux pump activity are yet to be identified.

  13. Regulation and production of Tcf, a cable-like fimbriae from Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Leclerc, Jean-Mathieu; Quevillon, Eve-Lyne; Houde, Yoan; Paranjape, Kiran; Dozois, Charles M; Daigle, France

    2016-05-01

    tcf (Typhi colonization factor) is one of the 12 putative chaperone/usher fimbrial clusters present in the Salmonella enterica serovar Typhi genome. We investigated the production, expression and regulation of tcf as well as its role during interaction with human cells. The tcf gene cluster was cloned and induced in Escherichia coli and S. Typhi, and the production of intertwined fibres similar to the Cbl (cable) pili of Burkholderia cepacia was observed on the bacterial surface by electron microscopy. In S. Typhi, tcf was expressed more after growth in M63 minimal medium than in standard Luria-Bertani medium. Analysis of the promoter region identified putative binding sites for the global regulators RcsB, ArgR and Fur. The expression of tcf was measured in isogenic strains lacking these global regulators. Under the conditions tested, the results showed that tcf expression was higher in the fur mutant and was regulated by iron concentration. Fur may regulate these fimbriae indirectly via the small RNAs RyhB1 and RyhB2. An isogenic mutant harbouring a deletion of the tcf cluster did not demonstrate any defect in adhesion or invasion of human epithelial cells, or in phagocytosis or survival in macrophages, when compared to the WT serovar Typhi strain. However, the tcf cluster contributed to adherence to human epithelial cells when introduced into E. coli. Thus, tcf genes encode functional fimbriae that can act as an adhesin and may contribute to colonization during typhoid fever. PMID:26944792

  14. SOS System Induction Inhibits the Assembly of Chemoreceptor Signaling Clusters in Salmonella enterica.

    Science.gov (United States)

    Irazoki, Oihane; Mayola, Albert; Campoy, Susana; Barbé, Jordi

    2016-01-01

    Swarming, a flagellar-driven multicellular form of motility, is associated with bacterial virulence and increased antibiotic resistance. In this work we demonstrate that activation of the SOS response reversibly inhibits swarming motility by preventing the assembly of chemoreceptor-signaling polar arrays. We also show that an increase in the concentration of the RecA protein, generated by SOS system activation, rather than another function of this genetic network impairs chemoreceptor polar cluster formation. Our data provide evidence that the molecular balance between RecA and CheW proteins is crucial to allow polar cluster formation in Salmonella enterica cells. Thus, activation of the SOS response by the presence of a DNA-injuring compound increases the RecA concentration, thereby disturbing the equilibrium between RecA and CheW and resulting in the cessation of swarming. Nevertheless, when the DNA-damage decreases and the SOS response is no longer activated, basal RecA levels and thus polar cluster assembly are reestablished. These results clearly show that bacterial populations moving over surfaces make use of specific mechanisms to avoid contact with DNA-damaging compounds.

  15. Antagonistic activity of Lactobacillus acidophilus LA10 against Salmonella enterica serovar Enteritidis SE86 in mice

    Directory of Open Access Journals (Sweden)

    Diane Scapin

    2013-01-01

    Full Text Available Salmonella enterica serovar Enteritidis is one of the main pathogens responsible for foodborne illness in Brazil. Probiotic bacteria can play a role in defense and recovery from enteropathogenic -infections. In this study, the ability of Lactobacillus acidophilus LA10 to colonise and exert anta-gonistic effects in the gastrointestinal tract was tested before and during experimental infection in conventional mice contaminated with S. Enteritidis (SE86. A dose of 0.1 mL containing 10(8 viable cells of SE86 and L. acidophilus LA10 was orally administered by gavage to mice. The experiment was divided into groups. As a negative control, Group 1 was administered only sterile saline solution. As a positive control, Group 2 was administered only SE86. Group 3 was first administered SE86, and after 10 days, treated with L. acidophilus LA10. Group 4 was first administered L. acidophilus LA10,and after 10 days, challenged with SE86.The results demonstrated that a significant number of SE86 cells were able to colonize the gastrointestinal tract of mice, specifically in the colon and ileum. L. acidophilus LA10 demonstrated an antagonistic effect against SE86, with better results observed for Group 3 over Group 4. Thus, L. acidophilus LA10 shows potential antagonistic effects against S. Enteritidis SE86, especially if administered after infection.

  16. Differential induction of total IgE by two Salmonella enterica serotypes

    Directory of Open Access Journals (Sweden)

    Zhanna eKtsoyan

    2015-06-01

    Full Text Available The main goal of this study was to establish how the inflammation caused by infection with two different Salmonella enterica serotypes, S. Typhimurium and S. Enteritidis, may lead to the predisposition to allergy as measured by total IgE level in the blood. Infection by S. Typhimurium did not affect the systemic IgE concentration while in S. Enteritidis-infected patients there was a significant 3.5-fold increase. This effect was especially profound in patients >4 years old, with up to the eight-fold increase above the norm. The degree of dysbiosis in these two infections measured with the comparative counts of cultivated bacteria showed an inverse relationship with the IgE concentration. Earlier we reported the elevated level of IL-17 in patients infected by S. Enteritidis. In the current study a significant correlation was found between the concentrations of IL-17 and IgE suggesting a possible role played by this cytokine in triggering the production of IgE in response to S. Enteritidis infection.

  17. Immunological evaluation of Vi capsular polysaccharide of Salmonella enterica subsp. Typhi vaccine by serum bactericidal assay.

    Science.gov (United States)

    Ahmadi, H; Tabaraie, B; Maleknia, S; Shapouri, R; Nejati, M; Pour Mirza Gholi, F; Hedayati, M; Sadati, M; Zahednia, S; Sharifat Salmani, A

    2013-02-01

    Salmonella enterica subsp. Typhi (S. Typhi) Vi antigen capsular polysaccharide (Vi-CPS) is a licensed vaccine against typhoid fever. As there is no animal model for S. Typhi fever to evaluate the protective efficacy of the Vi-CPS vaccine, a serum bactericidal assay (SBA) is the recommended 'gold standard' to evaluate its potency. Vi-CPS was extracted from S. Typhi Ty6S (CSBPI-B191) using a modified Gotschlich method. Purified Vi-CPS (50 µg) was injected intramuscularly into three groups of five rabbits; group 2 received an additional booster dose of 50 µg Vi-CPS on day 15 and group 3 received two additional boosters on days 15 and 30. The sera obtained from each group were tested by SBA on days 0, 15, 30 and 45. The anti-Vi-CPS titres for groups 1, 2 and 3 on days 15, 30 and 45 were 4, 16 and 16; 4, 32 and 32; and 16, 64 and 64, respectively. Thus, Vi-CPS was shown to be a potent immunogen, as even one dose could induce an efficient bactericidal effect against S. Typhi. Although Vi-CPS is a reliable vaccine, sometimes depolymerization during purification can affect its potency, which can be resolved through a potency test. As the passive haemagglutination test recommended by the World Health Organization does not indicate vaccine potency, we recommend using an SBA to evaluate the bactericidal ability of Vi-CPS.

  18. Regulation and production of Tcf, a cable-like fimbriae from Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Leclerc, Jean-Mathieu; Quevillon, Eve-Lyne; Houde, Yoan; Paranjape, Kiran; Dozois, Charles M; Daigle, France

    2016-05-01

    tcf (Typhi colonization factor) is one of the 12 putative chaperone/usher fimbrial clusters present in the Salmonella enterica serovar Typhi genome. We investigated the production, expression and regulation of tcf as well as its role during interaction with human cells. The tcf gene cluster was cloned and induced in Escherichia coli and S. Typhi, and the production of intertwined fibres similar to the Cbl (cable) pili of Burkholderia cepacia was observed on the bacterial surface by electron microscopy. In S. Typhi, tcf was expressed more after growth in M63 minimal medium than in standard Luria-Bertani medium. Analysis of the promoter region identified putative binding sites for the global regulators RcsB, ArgR and Fur. The expression of tcf was measured in isogenic strains lacking these global regulators. Under the conditions tested, the results showed that tcf expression was higher in the fur mutant and was regulated by iron concentration. Fur may regulate these fimbriae indirectly via the small RNAs RyhB1 and RyhB2. An isogenic mutant harbouring a deletion of the tcf cluster did not demonstrate any defect in adhesion or invasion of human epithelial cells, or in phagocytosis or survival in macrophages, when compared to the WT serovar Typhi strain. However, the tcf cluster contributed to adherence to human epithelial cells when introduced into E. coli. Thus, tcf genes encode functional fimbriae that can act as an adhesin and may contribute to colonization during typhoid fever.

  19. Interaction of graphene family materials with Listeria monocytogenes and Salmonella enterica

    Science.gov (United States)

    Kurantowicz, Natalia; Sawosz, Ewa; Jaworski, Sławomir; Kutwin, Marta; Strojny, Barbara; Wierzbicki, Mateusz; Szeliga, Jacek; Hotowy, Anna; Lipińska, Ludwika; Koziński, Rafał; Jagiełło, Joanna; Chwalibog, André

    2015-01-01

    Graphene family materials have unique properties, which make them valuable for a range of applications. The antibacterial properties of graphene have been reported; however, findings have been contradictory. This study reports on the antimicrobial proprieties of three different graphene materials (pristine graphene (pG), graphene oxide (GO), and reduced graphene oxide (rGO)) against the food-borne bacterial pathogens Listeria monocytogenes and Salmonella enterica. A high concentration (250 μg/mL) of all the analyzed graphenes completely inhibited the growth of both pathogens, despite their difference in bacterial cell wall structure. At a lower concentration (25 μg/mL), similar effects were only observed with GO, as growth inhibition decreased with pG and rGO at the lower concentration. Interaction of the nanoparticles with the pathogenic bacteria was found to differ depending on the form of graphene. Microscopic imaging demonstrated that bacteria were arranged at the edges of pG and rGO, while with GO, they adhered to the nanoparticle surface. GO was found to have the highest antibacterial activity.

  20. Identification of in vivo induced protein antigens of Salmonella enterica serovar Typhi during human infection

    Institute of Scientific and Technical Information of China (English)

    HU Yong; CONG YanGuang; LI Shu; RAO XianCai; WANG Gang; HU FuQuan

    2009-01-01

    During infectious disease episodes, pathogens express distinct subsets of virulence factors which allow them to adapt to different environments. Hence, genes that are expressed or upregulated in vivo are implicated in pathogenesis. We used in vivo induced antigen technology (IVIAT) to identify antigens which are expressed during infection with Salmonella enterica serovar Typhi. We identified 7 in vivo induced (IVI) antigens, which included BcfD (a fimbrial structural subunit), GrxC (a giutaredoxin 3),SapB (an ABC-type transport system), T3663 (an ABC-type uncharacterized transport system), T3816 (a putative rhodanese-related sulfurtransferase), T1497 (a probable TonB-dependent receptor) and T3689 (unknown function). Of the 7 identified antigens, 5 antigens had no cross-immunoreactivity in adsorbed control sera from healthy subjects. These 5 included BcfD, GrxC, SapB, T3663 and T3689. Antigens identified in this study are potential targets for drug and vaccine development and may be utilized as diagnostic agents.

  1. BIOCHEMICAL AND MOLECULAR CHARACTERIZATION OF SALMONELLA-ENTERICA SEROVAR BERTA, AND COMPARISON OF METHODS FOR TYPING

    DEFF Research Database (Denmark)

    Olsen, J. E.; Brown, D. J.; Baggesen, Dorte Lau;

    1992-01-01

    Strains of Salmonella enterica serovar berta (S. berta) from Denmark and seven other countries have been characterized with the aim of developing a rational typing strategy in connection with outbreak investigations, Biotyping divided the strains into H2S-positive (90 %) and H2S-negative (10 %) b...... with restriction enzyme analysis of plasmids seemed to be the most rational typing strategy for S. berta. The results indicated that S. berta strains regardless of geographical source or host are possibly clonal in nature....... %) biovars. Six percent of the strains were resistant to one or more antimicrobial agents. Eighty-eight percent of the strains carried plasmids and 52 different plasmid profiles were recognized. Six of the common plasmid sizes in these profiles were shown by restriction enzyme analyses to contain more than...... one plasmid species. More than 90 % of the strains had the same ribotype with the restriction enzymes Sma I and EcoR I and the same whole cell protein profile. Outer membrane protein profiles and isoenzyme profiles were identical in all S. berta analysed. Plasmid profiling in combination...

  2. Antioxidant oils and Salmonella enterica Typhimurium reduce tumor in an experimental model of hepatic metastasis

    Directory of Open Access Journals (Sweden)

    Sorenson BS

    2011-05-01

    Full Text Available Brent S Sorenson, Kaysie L Banton, Lance B Augustin, Arnold S Leonard, Daniel A SaltzmanDepartment of Surgery, University of Minnesota Medical School, Minneapolis, MN, USAAbstract: Fruit seeds high in antioxidants have been shown to have anticancer properties and enhance host protection against microbial infection. Recently we showed that a single oral dose of Salmonella enterica serovar Typhimurium expressing a truncated human interleukin-2 gene (SalpIL2 is avirulent, immunogenic, and reduces hepatic metastases through increased natural killer cell populations in mice. To determine whether antioxidant compounds enhance the antitumor effect seen in SalpIL2-treated animals, we assayed black cumin (BC, black raspberry (BR, and milk thistle (MT seed oils for the ability to reduce experimental hepatic metastases in mice. In animals without tumor, BC and BR oil diets altered the kinetics of the splenic lymphocyte response to SalpIL2. Consistent with previous reports, BR and BC seed oils demonstrated independent antitumor properties and moderate adjuvant potential with SalpIL2. MT oil, however, inhibited the efficacy of SalpIL2 in our model. Based on these data, we conclude that a diet high in antioxidant oils promoted a more robust immune response to SalpIL2, thus enhancing its antitumor efficacy.Keywords: antioxidants, colorectal cancer, tumor models, metastasis

  3. Effect of the Surfactant Tween 80 on the Detachment and Dispersal of Salmonella enterica Serovar Thompson Single Cells and Aggregates from Cilantro Leaves as Revealed by Image Analysis

    OpenAIRE

    Maria T Brandl; Huynh, Steven

    2014-01-01

    Salmonella enterica has the ability to form biofilms and large aggregates on produce surfaces, including on cilantro leaves. Aggregates of S. enterica serovar Thompson that remained attached to cilantro leaves after rigorous washing and that were present free or bound to dislodged leaf tissue in the wash suspension were observed by confocal microscopy. Measurement of S. Thompson population sizes in the leaf washes by plate counts failed to show an effect of 0.05% Tween 80 on the removal of th...

  4. Comparison of Methods of Extracting Salmonella enterica Serovar Enteritidis DNA from Environmental Substrates and Quantification of Organisms by Using a General Internal Procedural Control

    OpenAIRE

    Klerks, M. M.; Bruggen, van, C.; Zijlstra, C.; Donnikov, M.; Vos

    2006-01-01

    This paper compares five commercially available DNA extraction methods with respect to DNA extraction efficiency of Salmonella enterica serovar Enteritidis from soil, manure, and compost and uses an Escherichia coli strain harboring a plasmid expressing green fluorescent protein as a general internal procedural control. Inclusion of this general internal procedural control permitted more accurate quantification of extraction and amplification of S. enterica serovar Enteritidis in these sample...

  5. Emergence of Extended-Spectrum-β-Lactamase (CTX-M-9)-Producing Multiresistant Strains of Salmonella enterica Serotype Virchow in Poultry and Humans in France

    OpenAIRE

    Weill, François-Xavier; Lailler, Renaud; Praud, Karine; Kérouanton, Annaëlle; Fabre, Laëtitia; Brisabois, Anne; Grimont, Patrick A. D.; Cloeckaert, Axel

    2004-01-01

    During 2002 to 2003, eight Salmonella enterica serotype Virchow poultry and poultry product isolates from various sources (chicken farms, poultry slaughterhouse, or retail store) and one S. enterica rough strain isolated from human feces were found to produce extended-spectrum β-lactamase CTX-M-9. Poultry and poultry product isolates were recovered from different locations in the southwest of France. The human rough isolate had sequences of flagellin genes (fliC and fljB) typical of serotype ...

  6. Genomic Comparison of Non-Typhoidal Salmonella enterica Serovars Typhimurium, Enteritidis, Heidelberg, Hadar and Kentucky Isolates from Broiler Chickens.

    Directory of Open Access Journals (Sweden)

    Akhilesh S Dhanani

    Full Text Available Non-typhoidal Salmonella enterica serovars, associated with different foods including poultry products, are important causes of bacterial gastroenteritis worldwide. The colonization of the chicken gut by S. enterica could result in the contamination of the environment and food chain. The aim of this study was to compare the genomes of 25 S. enterica serovars isolated from broiler chicken farms to assess their intra- and inter-genetic variability, with a focus on virulence and antibiotic resistance characteristics.The genomes of 25 S. enterica isolates covering five serovars (ten Typhimurium including three monophasic 4,[5],12:i:, four Enteritidis, three Hadar, four Heidelberg and four Kentucky were sequenced. Most serovars were clustered in strongly supported phylogenetic clades, except for isolates of serovar Enteritidis that were scattered throughout the tree. Plasmids of varying sizes were detected in several isolates independently of serovars. Genes associated with the IncF plasmid and the IncI1 plasmid were identified in twelve and four isolates, respectively, while genes associated with the IncQ plasmid were found in one isolate. The presence of numerous genes associated with Salmonella pathogenicity islands (SPIs was also confirmed. Components of the type III and IV secretion systems (T3SS and T4SS varied in different isolates, which could explain in part, differences of their pathogenicity in humans and/or persistence in broilers. Conserved clusters of genes in the T3SS were detected that could be used in designing effective strategies (diagnostic, vaccination or treatments to combat Salmonella. Antibiotic resistance genes (CMY, aadA, ampC, florR, sul1, sulI, tetAB, and srtA and class I integrons were detected in resistant isolates while all isolates carried multidrug efflux pump systems regardless of their antibiotic susceptibility profile.This study showed that the predominant Salmonella serovars in broiler chickens harbor genes

  7. Orquiepididimite em carneiro por Salmonella enterica sub-diarizonae: primeiro caso na América do Sul Orchiepididymitis in ram by Salmonella enterica sub diarizonae: first case in South America

    Directory of Open Access Journals (Sweden)

    E.C.C. Celeghini

    2013-02-01

    Full Text Available O objetivo deste artigo é relatar um caso de orquiepididimite associado com infecção por Salmonella enterica subespécie diarizonae em carneiro da raça Santa Inês de quatro anos de idade, vasectomizado. Ao exame clínico reprodutivo, o animal mostrou aumento severo do conteúdo escrotal, sendo o testículo direito maior do que o esquerdo e a cauda do epidídimo direita maior do que a esquerda. A consistência testicular, avaliada em escala de 1 a 5, foi 5 para o testículo direito e 2,5 para o esquerdo; o órgão apresentava-se muito sensível ao toque. Na ultrassonografia foram observadas estruturas anecoicas/hipoecoicas circulares na cauda do epidídimo, sugestivas de abscessos; alguns pontos hiperecogênicos no parênquima testicular, sugerindo lesões de calcificação; e todo o testículo direito rodeado por imagem hipoecoica, indicativa de edema. Uma das estruturas da cauda do epidídimo direita foi puncionada, encontrando-se exsudato purulento, o qual foi enviado para exame microbiológico, sendo isolada e identificada Salmonella enterica subespécie diarizonae. O carneiro foi submetido a orquiepididectomia, e o órgão foi caracterizado macroscopicamente por adesões fibrosas entre as camadas escrotais, coexistência de abscessos epididimários e degeneração testicular. A Salmonella enterica subespécie diarizonae deve ser considerada no diagnóstico diferencial de infecção genital em ovinos.The objective of this manuscript was to report a case of orchiepididymitis associated with Salmonella enterica subespécie diarizonae infection in a vasectomized 4-year-old Santa Inês ram. In the clinical-reproductive examination, the animal showed a severe enlargement of the scrotal contents, being the right testicle larger than left, and the right epididymal cauda was higher than the left. The testicular consistency, evaluated in a scale from 1 to 5, was 5 to the right and 2.5 to the left, and the organ was very sensitive to the touch. In

  8. The two murein lipoproteins of Salmonella enterica serovar Typhimurium contribute to the virulence of the organism.

    Science.gov (United States)

    Sha, J; Fadl, A A; Klimpel, G R; Niesel, D W; Popov, V L; Chopra, A K

    2004-07-01

    Septic shock due to Salmonella and other gram-negative enteric pathogens is a leading cause of death worldwide. The role of lipopolysaccharide in sepsis is well studied; however, the contribution of other bacterial outer membrane components, such as Braun (murein) lipoprotein (Lpp), is not well defined. The genome of Salmonella enterica serovar Typhimurium harbors two copies of the lipoprotein (lpp) gene. We constructed a serovar Typhimurium strain with deletions in both copies of the lpp gene (lpp1 and lpp2) by marker exchange mutagenesis. The integrity of the cell membrane and the secretion of the effector proteins through the type III secretion system were not affected in the lpp double-knockout mutant. Subsequently, the virulence potential of this mutant was examined in a cell culture system using T84 intestinal epithelial and RAW264.7 macrophage cell lines and a mouse model of salmonellosis. The lpp double-knockout mutant was defective in invading and inducing cytotoxic effects in T84 and RAW264.7 cells, although binding of the mutant to the host cell was not affected when compared to the wild-type (WT) serovar Typhimurium. The motility of the mutant was impaired, despite the finding that the number of flagella was similar in the lpp double knockout mutant and the WT serovar Typhimurium. Deletion in the lpp genes did not affect the intracellular survival and replication of Salmonella in macrophages and T84 cells. Induction of the proinflammatory cytokines tumor necrosis factor alpha and interleukin-8 (IL-8) was significantly reduced in macrophages and T84 cells infected with the lpp double-knockout mutant. The levels of IL-8 remained unaffected in T84 cells when infected with either live or heat-killed WT and lpp mutant, indicating that invasion was not required for IL-8 production and that Toll-like receptor 2 signaling might be affected in the Lpp double-knockout mutant. These effects of the Lpp protein could be restored by complementation of the isogenic

  9. Genetic Transplantation: Salmonella enterica Serovar Typhimurium as a Host To Study Sigma Factor and Anti-Sigma Factor Interactions in Genetically Intractable Systems

    OpenAIRE

    Karlinsey, Joyce E.; Hughes, Kelly T.

    2006-01-01

    In Salmonella enterica serovar Typhimurium, σ28 and anti-sigma factor FlgM are regulatory proteins crucial for flagellar biogenesis and motility. In this study, we used S. enterica serovar Typhimurium as an in vivo heterologous system to study σ28 and anti-σ28 interactions in organisms where genetic manipulation poses a significant challenge due to special growth requirements. The chromosomal copy of the S. enterica serovar Typhimurium σ28 structural gene fliA was exchanged with homologs of A...

  10. Efficacy of several vaccination programmes in commercial layer and broiler breeder hens against experimental challenge with Salmonella enterica serovar Enteritidis.

    Science.gov (United States)

    Penha Filho, Rafael Antonio Casarin; de Paiva, Jacqueline Boldrin; Arguello, Yuli Melisa Sierra; da Silva, Mariana Dias; Gardin, Yannick; Resende, Fernando; Berchieri Junior, Angelo Berchieri; Sesti, Luiz

    2009-10-01

    Two experiments were performed to evaluate the protective effect of various vaccination combinations given at 5 and 9 weeks of age against experimental challenge with Salmonella enterica serovar Enteritidis (SE) phage type 4 at 12 weeks of age. In Experiment 1, groups of commercial layers were vaccinated by one of the following programmes: Group 1, two doses of a SE bacterin (Layermune SE); Group 2, one dose of a live Salmonella enterica serovar Gallinarum vaccine (Cevac SG9R) followed by one dose of the SE bacterin; Group 3, one dose of each of two different multivalent inactivated vaccines containing SE cells (Corymune 4K and Corymune 7K; and Group 4, unvaccinated, challenged controls. In Experiment 2, groups of broiler breeders were vaccinated by the same programmes as Groups 1 and 2 above while Group 3 was an unvaccinated, challenged control group. All vaccination programmes and the challenge induced significant (P liver and caeca) colonization by the challenge strain as well as reducing faecal excretion for at least 3 weeks. Vaccinated layers in Groups 1 and 2 and broiler breeders in Group 2 showed the greatest reduction in organ colonization and the least faecal excretion. In Experiment 1, layers vaccinated with multivalent inactivated vaccines containing a SE component (Group 3) were only moderately protected, indicating that such a vaccination programme may be useful in farms with good husbandry and housing conditions and low environmental infectious pressure by Salmonella.

  11. Excision of an unstable pathogenicity island in Salmonella enterica serovar Enteritidis is induced during infection of phagocytic cells.

    Directory of Open Access Journals (Sweden)

    Tania S Quiroz

    Full Text Available The availability of the complete genome sequence of several Salmonella enterica serovars has revealed the presence of unstable genetic elements in these bacteria, such as pathogenicity islands and prophages. This is the case of Salmonella enterica serovar Enteritidis (S. Enteritidis, a bacterium that causes gastroenteritis in humans and systemic infection in mice. The whole genome sequence analysis for S. Enteritidis unveiled the presence of several genetic regions that are absent in other Salmonella serovars. These regions have been denominated "regions of difference" (ROD. In this study we show that ROD21, one of such regions, behaves as an unstable pathogenicity island. We observed that ROD21 undergoes spontaneous excision by two independent recombination events, either under laboratory growth conditions or during infection of murine cells. Importantly, we also found that one type of excision occurred at higher rates when S. Enteritidis was residing inside murine phagocytic cells. These data suggest that ROD21 is an unstable pathogenicity island, whose frequency of excision depends on the environmental conditions found inside phagocytic cells.

  12. Excision of an unstable pathogenicity island in Salmonella enterica serovar Enteritidis is induced during infection of phagocytic cells.

    Science.gov (United States)

    Quiroz, Tania S; Nieto, Pamela A; Tobar, Hugo E; Salazar-Echegarai, Francisco J; Lizana, Rodrigo J; Quezada, Carolina P; Santiviago, Carlos A; Araya, Daniela V; Riedel, Claudia A; Kalergis, Alexis M; Bueno, Susan M

    2011-01-01

    The availability of the complete genome sequence of several Salmonella enterica serovars has revealed the presence of unstable genetic elements in these bacteria, such as pathogenicity islands and prophages. This is the case of Salmonella enterica serovar Enteritidis (S. Enteritidis), a bacterium that causes gastroenteritis in humans and systemic infection in mice. The whole genome sequence analysis for S. Enteritidis unveiled the presence of several genetic regions that are absent in other Salmonella serovars. These regions have been denominated "regions of difference" (ROD). In this study we show that ROD21, one of such regions, behaves as an unstable pathogenicity island. We observed that ROD21 undergoes spontaneous excision by two independent recombination events, either under laboratory growth conditions or during infection of murine cells. Importantly, we also found that one type of excision occurred at higher rates when S. Enteritidis was residing inside murine phagocytic cells. These data suggest that ROD21 is an unstable pathogenicity island, whose frequency of excision depends on the environmental conditions found inside phagocytic cells.

  13. [Phenotypic and molecular characteristics of Salmonella enterica serotype 1,4, [5] ,12:i:--a review].

    Science.gov (United States)

    Yang, Xiaojuan; Wu, Qingping; Zhang, Jumei; Guo, Weipeng

    2014-11-01

    Salmonella enterica serotype 1, 4, [5 ], 12: i:- ( Salmonella 1, 4, [5], 12: i:-), an emerging serotype antigenically related to Salmonella Typhimurium (1,4, [5], 12 : i:1,2) but lacking the second phase flagellar antigen, has been frequently detected in many countries over the last 10 years. Nowadays it seems to be one of the major serotypes responsible for human salmonellosis cases worldwide. In addition, multidrug resistance is quite common in Salmonella 1, 4, [5],12:i:-, the two major clones (labelled as Spanish and European clones) show multidrug resistance to four or more unrelated classes of antimicrobials mediated by plasmids or chromosome. Some resistance determinants including bla(TEM), bla(CTX-M(-1), aac(3)-IV, aadA2, cmlA1, sul1, sul2, dfrA12, strA-strB, tet (A) and tet (B) have been found in these multidrug resistance strains. The genomic characterization of 1,4, [5] ,12:i:- isolates suggests that this serovar is likely to gather several clones or strains that have independently emerged from S. Typhimurium, and have changed through multiple independent events involving different clonal groups. In later study, emphasis should be paid on development of rapid and precise detection methods and study of pathogenic and resistance mechanisms of Salmonella 1,4, [5] ,12:i:-.

  14. The combination of CRISPR-MVLST and PFGE provides increased discriminatory power for differentiating human clinical isolates of Salmonella enterica subsp. enterica serovar Enteritidis.

    Science.gov (United States)

    Shariat, Nikki; DiMarzio, Michael J; Yin, Shuang; Dettinger, Lisa; Sandt, Carol H; Lute, James R; Barrangou, Rodolphe; Dudley, Edward G

    2013-05-01

    Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) is a major cause of foodborne salmonellosis. Rapid, efficient and accurate methods for identification are required to track specific strains of S. Enteritidis during outbreaks of human salmonellosis. By exploiting the hypervariable nature of virulence genes and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs), we previously developed a powerful sequence-based subtyping approach, designated CRISPR-MVLST. To substantiate the applicability of CRISPR-MVLST, we analyzed a broad set of S. Enteritidis isolates collected over a six-year period. Among 141 isolates we defined 22 Enteritidis Sequence Types (ESTs), the majority of which were novel. Notably, strains exhibiting the common PFGE pattern, JEGX01.0004 (characteristic of ∼40% of S. Enteritidis isolates in the United States), were separated into twelve distinct sequence types. Conversely, isolates of EST4, the most predominant EST we observed, comprised eight different PFGE patterns. Importantly, we showed that some genotypes that were previously associated with the food supply chain at the farm level have now been identified in clinical samples. CRISPR sequence data shows subtle but distinct differences among different alleles of S. Enteritidis, suggesting that evolution of these loci occurs vertically, as opposed to previously reported evolution by spacer acquisition in other bacteria.

  15. Aneurysm of the cranial mesenteric artery as a site of carriage of Salmonella enterica subsp. enterica serovar Abortusequi in the horse.

    Science.gov (United States)

    Niwa, Hidekazu; Hobo, Seiji; Kinoshita, Yuta; Muranaka, Masanori; Ochi, Akihiro; Ueno, Takanori; Oku, Kazuomi; Hariu, Kazuhisa; Katayama, Yoshinari

    2016-07-01

    Salmonella enterica subsp. enterica serovar Abortusequi is a pathogen restricted to horses. Our investigation targeted 4 draft horses (9-10 months old) kept on a Japanese farm that had suffered an outbreak of S. Abortusequi abortion. The 4 horses were suspected to be carriers of the bacterium owing to their high agglutination titers (≥1:2,560) in tube agglutination testing. The owners' on-farm observations confirmed that the horses had no apparent abnormalities, and S. Abortusequi was not isolated from their blood, rectal swabs, or sternal bone marrow fluid at antemortem investigation. However, at autopsy, all horses displayed the following: suppurative aneurysm of the cranial mesenteric artery with heavy infection with Strongylus vulgaris larvae; heavy intestinal parasitic infection with Gasterophilus intestinalis, Parascaris equorum, Anoplocephala perfoliata, and S. vulgaris; and enlargement of the systemic lymph nodes. In each case, large numbers of S. Abortusequi were isolated from the anterior mesenteric artery thrombus. The thrombus isolates harbored a single virulence plasmid, and the pulsed-field gel electrophoresis profiles of the isolates were identical not only to each other but also to those of Japanese enzootic strains of S. Abortusequi. These results reveal that parasitic aneurysms of the cranial mesenteric artery should be considered an important possible site of carriage of S. Abortusequi in horses. The results also suggest high clonality of the isolated serovar in the horse population in Japan. PMID:27271985

  16. A Lactobacillus rhamnosus strain induces protection in different sites after Salmonella enterica subsp. enterica serovar Typhimurium challenge in gnotobiotic and conventional mice

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    A.H. Mendonça

    2014-04-01

    Full Text Available The ability of a Lactobacillus rhamnosus strain isolated from a healthy breast-fed human newborn to reduce the pathological consequences for the host due to an experimental oral infection with Salmonella enterica subsp. enterica serov. Typhimurium in vivo was determined using gnotobiotic and conventional mice. Conventional mice received 0.1mL probiotic milk (8.0 log colony-forming unit daily for 10 days before the oral pathogenic challenge (5.0 log colony-forming unit. Then probiotic treatment was continued until the end of the experiment. Probiotic treatment in germ-free mice consisted of a single dose of the probiotic milk at the beginning of the experiment and a challenge with S. Typhimurium 10 days later (3.0 log colony-forming unit. A protective effect was observed in both gnotobiotic and conventional animals in terms of histopathologic and morphometric data, but in different anatomical sites. This protection was observed in liver and intestines, respectively, for gnotobiotic and conventional mice. However, S. Typhimurium populations were similar in the feces of both treated and control gnotobiotic mice. We conclude that a protective effect of L. rhamnosus against experimental S. Typhimurium was observed. This protection was not due to the reduction of the population of pathogenic bacteria in the intestine.

  17. Protection by Lactobacillus acidophilus UFV-H2B20 against experimental oral infection with Salmonella enterica subsp. enterica Ser. Typhimurium in gnotobiotic and conventional mice

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    Moura Lilian Nobre

    2001-01-01

    Full Text Available The ability of Lactobacillus acidophilus UFV-H2B20 to antagonize Salmonella enterica subsp. enterica ser. Typhimurium and to reduce the pathological consequences for the host was determining using conventional and gnotobiotic animals. Conventional NIH mice received daily by gavage a 0.1 ml suspension containing about 10(8 cfu L. acidophilus UFV-H2B20 and germfree animals received a single 0.1 ml dose. The gnotobiotic and conventional groups were infected orally with 10² and 10(5 cfu of S. Typhimurium, respectively, 7 days after the beginning of treatment. Control groups were treated with sterile saline instead of Lactobacillus. Survival data showed a protective effect against the pathogenic bacteria in both conventional and gnotobiotic Lactobacillus-treated mice. L. acidophilus UFV-H2B20 colonized the digestive tract of gnotobiotic mice and the number of viable cells ranged from 10(9 to 10(10 cfu/g of faeces. In both experimental and control gnotobiotic animals, S. Typhimurium became rapidly established at a level ranging from 10(8 to 10(10 cfu/g of faeces and remained at high levels until the animals died or were sacrificed. In conclusion, the previous treatment of mice with L. acidophilus UFV-H2B20 protects the animals against the experimental infection with S. Typhimurium but this protection was not due to the reduction of the pathogenic populations in the intestines.

  18. Complex Function for SicA, a Salmonella enterica Serovar Typhimurium Type III Secretion-Associated Chaperone

    OpenAIRE

    Tucker, Stephanie C.; Galán, Jorge E.

    2000-01-01

    Salmonella enterica encodes a type III secretion system within a pathogenicity island located at centisome 63 that is essential for virulence. All type III secretion systems require the function of a family of low-molecular-weight proteins that aid the secretion process by acting as partitioning factors and/or secretion pilots. One such protein is SicA, which is encoded immediately upstream of the type III secreted proteins SipB and SipC. We found that the absence of SicA results in the degra...

  19. Molecular and epidemiologic analysis of a county-wide outbreak caused by Salmonella enterica subsp. enterica serovar Enteritidis traced to a bakery

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    Lu Po-Liang

    2004-11-01

    Full Text Available Abstract Background An increase in the number of attendees due to acute gastroenteritis and fever was noted at one hospital emergency room in Taiwan over a seven-day period from July to August, 2001. Molecular and epidemiological surveys were performed to trace the possible source of infection. Methods An epidemiological investigation was undertaken to determine the cause of the outbreak. Stool and blood samples were collected according to standard protocols per Center for Disease Control, Taiwan. Typing of the Salmonella isolates from stool, blood, and food samples was performed with serotyping, antibiotypes, and pulsed field gel electrophoresis (PFGE following XbaI restriction enzyme digestion. Results Comparison of the number of patients with and without acute gastroenteritis (506 and 4467, respectively during the six weeks before the outbreak week revealed a significant increase in the number of patients during the outbreak week (162 and 942, respectively (relative risk (RR: 1.44, 95% confidence interval (CI: 1.22–1.70, P value Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis was isolated from the stool samples of 28 of 32 individuals and from a recalled bread sample. All S. Enteritidis isolates were of the same antibiogram. PFGE typing revealed that all except two of the clinical isolates and the bread isolates were of the same DNA macrorestriction pattern. Conclusions The egg-covered bread contaminated with S. Enteritidis was confirmed as the vehicle of infection. Alertness in the emergency room, surveillance by the microbiology laboratory, prompt and thorough investigation to trace the source of outbreaks, and institution of appropriate control measures provide effective control of community outbreaks.

  20. 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 cells (P cells at 14 dpi (P cells (P cells (P Salmonella Typhimurium in comparison with Salmonella Typhimurium challenge alone. β-1,3/1,6-Glucan addition had significant inhibitory effects (P Salmonella colonization levels and liver Salmonella invasion of the Salmonella Typhimurium-infected birds compared with the PC group. Intestinal tight junction proteins claudin-1, claudin-4, and occludin mRNA expression in the jejunum at 14 dpi was significantly decreased by Salmonella Typhimurium challenge alone (P Salmonella Typhimurium-infected birds in comparison with the PC group. Our results indicate that dietary β-1,3/1,6-glucan can alleviate intestinal mucosal barrier impairment in broiler chickens challenged with Salmonella Typhimurium.

  1. Specific responses of Salmonella enterica to tomato varieties and fruit ripeness identified by in vivo expression technology.

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    Jason T Noel

    Full Text Available BACKGROUND: Recent outbreaks of vegetable-associated gastroenteritis suggest that enteric pathogens colonize, multiply and persist in plants for extended periods of time, eventually infecting people. Genetic and physiological pathways, by which enterics colonize plants, are still poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: To better understand interactions between Salmonella enterica sv. Typhimurium and tomatoes, a gfp-tagged Salmonella promoter library was screened inside red ripe fruits. Fifty-one unique constructs that were potentially differentially regulated in tomato relative to in vitro growth were identified. The expression of a subset of these promoters was tested in planta using recombinase-based in vivo expression technology (RIVET and fitness of the corresponding mutants was tested. Gene expression in Salmonella was affected by fruit maturity and tomato cultivar. A putative fadH promoter was upregulated most strongly in immature tomatoes. Expression of the fadH construct depended on the presence of linoleic acid, which is consistent with the reduced accumulation of this compound in mature tomato fruits. The cysB construct was activated in the fruit of cv. Hawaii 7997 (resistant to a race of Ralstonia solanacearum more strongly than in the universally susceptible tomato cv. Bonny Best. Known Salmonella motility and animal virulence genes (hilA, flhDC, fliF and those encoded on the pSLT virulence plasmid did not contribute significantly to fitness of the bacteria inside tomatoes, even though deletions of sirA and motA modestly increased fitness of Salmonella inside tomatoes. CONCLUSIONS/SIGNIFICANCE: This study reveals the genetic basis of the interactions of Salmonella with plant hosts. Salmonella relies on a distinct set of metabolic and regulatory genes, which are differentially regulated in planta in response to host genotype and fruit maturity. This enteric pathogen colonizes tissues of tomatoes differently than plant

  2. Studies on the effects of phosphine on Salmonella enterica serotype Enteritidis in culture medium and in black pepper (Piper nigrum).

    Science.gov (United States)

    Castro, M F P M; Rezende, A C B; Benato, E A; Valentini, S R T; Furlani, R P Z; Tfouni, S A V

    2011-04-01

    The effect of phosphine on Salmonella enterica serotype Enteritidis inoculated in culture medium and in black pepper grains (Piper nigrum), as well as on the reduction of the microbial load of the dried and moisturized product, was verified. The postfumigation effect was verified in inoculated samples with 0.92 and 0.97 water activity (a(w)) exposed to 6 g/m(3) phosphine for 72 h, dried to 0.67 a(w), and stored for 24, 48, and 72 h. No decreases were observed in Salmonella Enteritidis populations in culture medium when fumigant concentrations up to 6 g/m(3) were applied for 48 h at 35°C. However, the colonies showed reductions in size and atypical coloration as the phosphine concentration increased. No reduction in Salmonella counts occurred on the inoculated dried samples after fumigation. On the other hand, when phosphine at concentrations of 6 g/m(3) was applied on moisturized black pepper for 72 h, decreases in Salmonella counts of around 80% were observed. The counts of total aerobic mesophilic bacterium populations of the dried and moisturized black pepper were not affected by the fumigant treatment. The results of the postfumigation studies indicated that Salmonella Enteritidis was absent in the fumigated grains after drying and storage for 72 h, indicating a promising application for this technique. It was concluded that for Salmonella Enteritidis control, phosphine fumigation could be applied to black pepper grains before drying and the producers should rigidly follow good agricultural practices, mainly during the drying process, in order to avoid product recontamination. Additional work is needed to confirm the findings with more Salmonella serotypes and strains.

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

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    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

  4. Characterization and differential gene expression between two phenotypic phase variants in Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Patterson, Sheila K; Borewicz, Klaudyna; Johnson, Timothy; Xu, Wayne; Isaacson, Richard E

    2012-01-01

    Salmonella enterica serovar Typhimurium strain 798 has previously been shown to undergo phenotypic phase variation. One of the phenotypes expresses virulence traits such as adhesion, while the other phenotype does not. Phenotypic phase variation appears to correlate with the ability of this strain to cause persistent, asymptomatic infections of swine. A new method to detect cells in either phenotypic phase was developed using Evans Blue-Uranine agar plates. Using this new assay, rates of phenotypic phase variation were obtained. The rate of phase variation from non-adhesive to adhesive phenotype was approximately 10(-4) per cell per generation while phase variation from the adhesive to the non-adhesive phenotype was approximately 10(-6) per cell per generation. Two highly virulent S. Typhimurium strains, SL1344 and ATCC 14028, were also shown to undergo phase variation. However, while the rate from adhesive to non-adhesive phenotype was approximately the same as for strain 798, the non-adhesive to adhesive phenotype shift was 37-fold higher. Differential gene expression was measured using RNA-Seq. Eighty-three genes were more highly expressed by 798 cells in the adhesive phenotype compared to the non-adhesive cells. Most of the up-regulated genes were in virulence genes and in particular all genes in the Salmonella pathogenicity island 1 were up-regulated. When compared to the virulent strain SL1344, expression of the virulence genes was approximately equal to those up-regulated in the adhesive phenotype of strain 798. A comparison of invasive ability demonstrated that strain SL1344 was the most invasive followed by the adhesive phenotype of strain 798, then the non-adhesive phenotype of strain 798. The least invasive strain was ATCC 14028. The genome of strain 798 was sequenced and compared to SL1344. Both strains had very similar genome sequences and gene deletions could not readily explain differences in the rates of phase variation from non-adhesive to the

  5. Characterization and differential gene expression between two phenotypic phase variants in Salmonella enterica serovar Typhimurium.

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    Sheila K Patterson

    Full Text Available Salmonella enterica serovar Typhimurium strain 798 has previously been shown to undergo phenotypic phase variation. One of the phenotypes expresses virulence traits such as adhesion, while the other phenotype does not. Phenotypic phase variation appears to correlate with the ability of this strain to cause persistent, asymptomatic infections of swine. A new method to detect cells in either phenotypic phase was developed using Evans Blue-Uranine agar plates. Using this new assay, rates of phenotypic phase variation were obtained. The rate of phase variation from non-adhesive to adhesive phenotype was approximately 10(-4 per cell per generation while phase variation from the adhesive to the non-adhesive phenotype was approximately 10(-6 per cell per generation. Two highly virulent S. Typhimurium strains, SL1344 and ATCC 14028, were also shown to undergo phase variation. However, while the rate from adhesive to non-adhesive phenotype was approximately the same as for strain 798, the non-adhesive to adhesive phenotype shift was 37-fold higher. Differential gene expression was measured using RNA-Seq. Eighty-three genes were more highly expressed by 798 cells in the adhesive phenotype compared to the non-adhesive cells. Most of the up-regulated genes were in virulence genes and in particular all genes in the Salmonella pathogenicity island 1 were up-regulated. When compared to the virulent strain SL1344, expression of the virulence genes was approximately equal to those up-regulated in the adhesive phenotype of strain 798. A comparison of invasive ability demonstrated that strain SL1344 was the most invasive followed by the adhesive phenotype of strain 798, then the non-adhesive phenotype of strain 798. The least invasive strain was ATCC 14028. The genome of strain 798 was sequenced and compared to SL1344. Both strains had very similar genome sequences and gene deletions could not readily explain differences in the rates of phase variation from non

  6. Growth and virulence properties of biofilm-forming Salmonella enterica serovar typhimurium under different acidic conditions.

    Science.gov (United States)

    Xu, Hua; Lee, Hyeon-Yong; Ahn, Juhee

    2010-12-01

    This study was designed to characterize the viability and potential virulence of bofilm-forming Salmonella enterica serovar Typhimurium under different pH levels, ranging from 5 to 7. The plate count method and real-time reverse transcription-PCR (RT-PCR) were used to evaluate the survival of S. Typhimurium grown in Trypticase soy broth (TSB) adjusted to pH 5, 6, and 7 (TSB-5, TSB-6, and TSB-7, respectively) at 37°C for 10 days. In TSB-5 and TSB-6, the numbers of viable cells estimated by using the real-time RT-PCR were greater than the culturable counts enumerated by the plate count method. Reflectance micro-Fourier transform infrared (micro-FTIR) spectroscopy was used to evaluate the biochemical changes in biofilm cells. Considerable changes in chemical components were observed in the biofilm cells grown in TSB-5 and TSB-6 when compared to the cells grown in TSB-7. The enterotoxin production and invasive ability of planktonic and biofilm S. Typhimurium cells were inferred by the relative levels of expression of stn and invA. The levels of expression of stn and invA were significantly increased in biofilm S. Typhimurium cells grown in TSB-5 (1.9-fold and 3.2-fold) and TSB-6 (2.1-fold and 22.3-fold) after 10 days of incubation. These results suggest that the biofilm-forming S. Typhimurium under different pH levels might change the virulence production and stress response mechanisms.

  7. Characterization of Novel Factors Involved in Swimming and Swarming Motility in Salmonella enterica Serovar Typhimurium.

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    Julia Andrea Deditius

    Full Text Available Salmonella enterica utilizes flagellar motility to swim through liquid environments and on surfaces. The biosynthesis of the flagellum is regulated on various levels, including transcriptional and posttranscriptional mechanisms. Here, we investigated the motility phenotype of 24 selected single gene deletions that were previously described to display swimming and swarming motility effects. Mutations in flgE, fliH, ydiV, rfaG, yjcC, STM1267 and STM3363 showed an altered motility phenotype. Deletions of flgE and fliH displayed a non-motile phenotype in both swimming and swarming motility assays as expected. The deletions of STM1267, STM3363, ydiV, rfaG and yjcC were further analyzed in detail for flagellar and fimbrial gene expression and filament formation. A ΔydiV mutant showed increased swimming motility, but a decrease in swarming motility, which coincided with derepression of curli fimbriae. A deletion of yjcC, encoding for an EAL domain-containing protein, increased swimming motility independent on flagellar gene expression. A ΔSTM1267 mutant displayed a hypermotile phenotype on swarm agar plates and was found to have increased numbers of flagella. In contrast, a knockout of STM3363 did also display an increase in swarming motility, but did not alter flagella numbers. Finally, a deletion of the LPS biosynthesis-related protein RfaG reduced swimming and swarming motility, associated with a decrease in transcription from flagellar class II and class III promoters and a lack of flagellar filaments.

  8. Isolation of Salmonella enterica serotype Worthington from a splenic abscess in a patient with chronic myeloid leukemia

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    Ghadage D.P.

    2002-01-01

    Full Text Available Splenic abscesses are caused by Staphylococcus aureus, Streptococcus and bacteria belonging to the family Enterobacteriaceae. We report a case of splenic abscess caused by an unusual serotype of Salmonella. A 55 year old man was admitted with complaints of fever and abdominal pain. On the basis of clinical findings and laboratory reports, a diagnosis of chronic myeloid leukemia was made. Ultrasonography of the abdomen revealed a single large cystic lesion in the spleen. Percutaneous drainage of the abscess was carried out. Salmonella enterica serotype Worthington was isolated from a pus sample taken from the abscess. The isolate was resistant to ampicillin, gentamicin, cefotaxime, chloramphenicol and tetracycline, and sensitive to amikacin and norfloxacin. Serotype Worthington is an emerging pathogen. This is the first report of isolation of this serotype from a splenic abscess. In seriously ill patients, such infections should be treated with a combination of antibiotics to circumvent problems with multidrug resistance.

  9. Emergence and clonal dissemination of Salmonella enterica serovar Enteritidis causing salmonellosis in Mauritius

    DEFF Research Database (Denmark)

    Issack, Mohammad I.; Hendriksen, Rene S.; Hyytiae-Trees, Eija;

    2014-01-01

    Introduction: For decades, Salmonella enterica serovar Enteritidis has been among the most prevalent serovars reported worldwide. However, it was rarely encountered in Mauritius until 2007; since then the number of non-typhoidal Salmonella serogroup O:9 (including serovar Enteritidis) increased. A...... food specimens were characterized by antimicrobial susceptibility testing and Multiple-Locus Variable-number tandem repeat Analysis (MLVA). Results: With the exception of a single isolate which demonstrated intermediate susceptibility to streptomycin, all isolates were pansusceptible to the 14...... closely related to blood and exudate isolates, indicating that, besides gastroenteritis, the same strain caused invasive infections. Control of S. Enteritidis by poultry breeders would lower the financial burden associated with morbidity in humans caused by this organism in Mauritius....

  10. Complete Genome Sequence of Salmonella enterica Serovar Typhimurium Strain SO2 (Sequence Type 302) Isolated from an Asymptomatic Child in Mexico

    Science.gov (United States)

    Calva, Edmundo; Puente, José L.; Zaidi, Mussaret B.

    2016-01-01

    The complete genome sequence of Salmonella enterica serovar Typhimurium strain SO2, isolated from an asymptomatic child in Mexico, was determined using PacBio single-molecule real-time technology. Strain SO2 has six complete chromosomal prophages, namely, ST104, Gifsy-2, ST64B, Gifsy-1, ELPhiS, and FSL SP-004, and carries a Salmonella virulence plasmid. PMID:27081133

  11. Pediatric Epidemic of Salmonella enterica Serovar Typhimurium in the Area of L’Aquila, Italy, Four Years after a Catastrophic Earthquake

    OpenAIRE

    Giovanni Nigro; Gabriella Bottone; Daniela Maiorani; Fabiana Trombatore; Silvana Falasca; Gianfranco Bruno

    2016-01-01

    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, inc...

  12. Salmonella enterica serovar Enteritidis brain abscess mimicking meningitis after surgery for glioblastoma multiforme: a case report and review of the literature

    OpenAIRE

    Luciani, Léa; Dubourg, Grégory; Graillon, Thomas; Honnorat, Estelle; Lepidi, Hubert; Drancourt, Michel; Seng, Piseth; Stein, Andreas

    2016-01-01

    Background Salmonella brain abscess associated with brain tumor is rare. Only 11 cases have been reported to date. Here we report a case of brain abscess caused by Salmonella enterica serovar Enteritidis mimicking post-surgical meningitis in a patient with glioblastoma multiforme. Case presentation A 60-year-old Algerian woman was admitted through an emergency department for a 4-day history of headache, nausea and vomiting, and behavioral disorders. Surgery for cerebral tumor excision was per...

  13. Genetic Fine Structure of a Salmonella enterica Serovar Typhi Strain Associated with the 2005 Outbreak of Typhoid Fever in Kelantan, Malaysia

    OpenAIRE

    Baddam, Ramani; Kumar, Narender; Thong, Kwai-Lin; Ngoi, Soo-Tein; Teh, Cindy Shuan Ju; Yap, Kien-Pong; Chai, Lay-Ching; Avasthi, Tiruvayipati Suma; Ahmed, Niyaz

    2012-01-01

    Among enteric pathogens, Salmonella enterica serovar Typhi is responsible for the largest number of food-borne outbreaks and fatalities. The ability of the pathogen to cause systemic infection for extended durations leads to a high cost of disease control. Chronic carriers play important roles in the evolution of Salmonella Typhi; therefore, identification and in-depth characterization of isolates from clinical cases and carriers, especially those from zones of endemicity where the pathogen h...

  14. Broilers with low serum Mannose-binding Lectin show increased fecal shedding of Salmonella enterica serovar Montevideo.

    Science.gov (United States)

    Ulrich-Lynge, Sofie L; Juul-Madsen, Helle R; Kjærup, Rikke B; Okimoto, Ron; Abrahamsen, Mitchell S; Maurischat, Sven; Sørensen, Poul; Dalgaard, Tina S

    2016-08-01

    Mannose-binding lectin (MBL) is a key molecule in innate immunity. MBL binds to carbohydrates on the surface of pathogens, initiating the complement system via the lectin-dependent pathway or facilitates opsonophagocytosis. In vivo studies using inbred chicken lines differing in MBL serum concentration indicate that chicken MBL affects Salmonella resistance; further studies are imperative in conventional broiler chickens. In this study 104 conventional day-old chickens (offspring from a cross between Cobb 500 male and female parent breeders) were orally infected with Salmonella enterica subsp. enterica serovar Montevideo. The chickens were divided into two groups based on polymorphisms in their MBL promoter region, designated L/L for low serum concentrations of MBL and L/H for medium serum concentrations of MBL. A semi-quantitative real-time PCR method for detection of Salmonella in cloacal swabs was used, the log10 CFU quantification was based on a standard curve from artificially spiked cloacal swab samples pre-incubated for 8 h with known concentrations of Salmonella ranging from 10(1) to 10(6) CFU/swabs, with an obtained amplification efficiency of 102% and a linear relationship between the log10 CFU and the threshold cycle Ct values of (R(2) = 0.99). The L/L chickens had significantly higher Log10 CFU/swab at week 5 post infection (pi) than the L/H chickens. A repetition of the study with 86 L/L and 18 L/H chickens, also gave significantly higher log10 CFU ± SEM in cloacal swabs, using the semi-quantitative real-time PCR method from L/L chickens than from the L/H chickens at week 5 pi. These results indicate that genetically determined basic levels of MBL may influence S. Montevideo susceptibility. PMID:26994208

  15. Tolerance response of multidrug-resistant Salmonella enterica strains to habituation to Origanum vulgare L essential oil

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    Daniel eMonte

    2014-12-01

    Full Text Available Multidrug-resistant Salmonella enterica isolates from human outbreaks or from poultry origin were investigated for their ability to develop direct-tolerance or cross-tolerance to sodium chloride (NaCl, potassium chloride (KCl, lactic acid (LA, acetic acid (AA and ciprofloxacin (CIP after habituation in subinhibitory amounts (½ of the minimum inhibitory concentration - ½ MIC and ¼ of the minimum inhibitory concentration - ¼ MIC of Origanum vulgare L. essential oil (OVEO at different time intervals. The habituation of S. enterica to OVEO did not induce direct-tolerance or cross-tolerance in the tested strains, as assessed by the modulation of minimum inhibitory concentration (MIC values. However, cells habituated to OVEO maintained or increased susceptibility to the tested antimicrobials agents, with up to four-fold double dilution decrease from previously determined MIC values. This study reports for the first time the non-inductive effect of OVEO on the acquisition of direct-tolerance or cross-tolerance in multidrug-resistant S. enterica strains to antimicrobial agents that are largely used in food preservation, as well as to CIP, the therapeutic drug of salmonellosis.

  16. A defective mutant of Salmonella enterica Serovar Gallinarum in cobalamin biosynthesis is avirulent in chickens Mutante de Salmonella enterica serovar Gallinarum duplo defectivo na biossíntese de cobalamina é avirulento para aves

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    Jacqueline Boldrin de Paiva

    2009-09-01

    Full Text Available Salmonella enterica serovar Gallinarum (SG is a fowl typhoid agent in chickens and is a severe disease with worldwide economic impact as its mortality may reach up to 80%. It is one of a small group of serovars that typically produces typhoid-like infections in a narrow range of host species and which therefore represents a good model for human typhoid. The survival mechanisms are not considered to be virulent mechanisms but are essential for the life of the bacterium. Mutants of Salmonella Gallinarum containing defective genes, related to cobalamin biosynthesis and which Salmonella spp. has to be produced to survive when it is in an anaerobic environment, were produced in this study. Salmonella Gallinarum is an intracellular parasite. Therefore, this study could provide information about whether vitamin B12 biosynthesis might be essential to its survival in the host. The results showed that the singular deletion in cbiA or cobS genes did not interfere in the life of Salmonella Gallinarum in the host, perhaps because single deletion is not enough to impede vitamin B12 biosynthesis. It was noticed that diluted SG mutants with single deletion produced higher mortality than the wild strain of SG. When double mutation was carried out, the Salmonella Gallinarum mutant was unable to provoke mortality in susceptible chickens. This work showed that B12 biosynthesis is a very important step in the metabolism of Salmonella Gallinarum during the infection of the chickens. Further research on bacterium physiology should be carried out to elucidate the events described in this research and to assess the mutant as a vaccine strain.Salmonella enterica serovar Gallinarum (SG é o agente do tifo aviário, doença severa que provoca mortalidade em até 80% do plantel de aves. SG encontra-se entre os poucos sorotipos de Salmonella que são agentes etiológicos de enfermidade específica, à semelhança de Salmonella Typhi em seres humanos podendo, portanto, servir

  17. Immuno-fluorescence based Vi capsular polysaccharide detection for specific recognition of Salmonella enterica serovar Typhi in clinical samples.

    Science.gov (United States)

    Pandey, Satish K; Vinayaka, Aaydha C; Rishi, Dharam B; Rishi, Praveen; Suri, C Raman

    2014-09-01

    Typhoid fever is a life threatening bacterial infection that remains a major global health concern. This continued high burden associated with significant morbidity and mortality rate demands specific and rapid detection technique. This work reports a new sandwich type fluorescence immunoassay format using polymyxin B, a cationic receptor molecule, as a binder agent while anti-Vi antibody served as the capturing agent for specifically detecting Salmonella enterica serovar Typhi. Anti-Vi IgG antibody raised against Vi-BSA conjugate revealed affinity of 7.779nM(-1) signifying immunodominancy of O-acetyls groups in Vi polysaccharide. The detection limit of the developed assay was around 10(1) cellsmL(-1) of Vi expressing Salmonella enterica serovar Typhi with a correlation coefficient (R(2)) equal to 0.97. Positive response obtained for all the tested serovar Typhi clinical isolates as well as the pathogen spiked blood samples recommended specificity and accuracy of Vi antigen as a biomarker during typhoid fever. The intra- and inter-assay precision with Vi spiked samples were satisfactory revealing coefficient of variance (CV%) with a mean of 4.05% and 5.97% respectively. This may be the novel attempt and constructive report on the fluorescence based detection of Vi antigen of serovar Typhi in the epidemic as well as pandemic outbreaks.

  18. Effect of X-ray treatments on Escherichia coli 0157:H7, Listeria monocytogenes, Shigella flexneri, Salmonella enterica and inherent microbiota on whole mangoes

    Science.gov (United States)

    The aims of this investigation were to; (i) study the effect of X-ray treatments in reducing Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica and Shigella flexneri on whole mangoes, and (ii) study the effect of Xray treatments on microflora counts (mesophilic counts, psychrotrop...

  19. Effect of the surfactant Tween 80 on the detachment and dispersal of Salmonella enterica Thompson single cells and aggregates from cilantro leaves as revealed by image analysis

    Science.gov (United States)

    Biofilms formed by human enteric pathogens on plants are a great concern to the produce industry. Salmonella enterica has the ability to form biofilms and large aggregates on leaf surfaces, including on cilantro leaves. Aggregates that remained attached after rigorous washing of cilantro leaves and ...

  20. Comparison of Methods of Extracting Salmonella enterica Serovar Enteritidis DNA from Environmental Substrates and Quantification of Organisms by Using a General Internal Procedural Control

    NARCIS (Netherlands)

    Klerks, M.M.; Bruggen, van A.H.C.; Zijlstra, C.; Donnikov, M.; Vos, de R.

    2006-01-01

    This paper compares five commercially available DNA extraction methods with respect to DNA extraction efficiency of Salmonella enterica serovar Enteritidis from soil, manure, and compost and uses an Escherichia coli strain harboring a plasmid expressing green fluorescent protein as a general interna

  1. The role of QseC quorum-sensing sensor kinase in colonization and norepinephrine-enhanced motility of Salmonella enterica serovar Typhirmurium

    Science.gov (United States)

    Transcriptional analysis of Salmonella enterica serovar Typhimurium in the presence of the mammalian hormone Norepinephrine (NE) revealed up-regulation of chemotaxis and motility genes. Motility assays confirmed enhanced motility of wild-type S. Typhimurium in the presence of NE that could be block...

  2. Chicken-specific kinome array reveals that Salmonella enterica serovar Enteritidis modulates host immune signaling pathways in the cecum to establish a persistence infection

    Science.gov (United States)

    Non-typhoidal Salmonella enterica induce an early, short-lived, pro-inflammatory response in chickens that is asymptomatic of clinical disease and results in a persistent colonization of the gastrointestinal (GI) tract that transmits infections to naïve hosts via fecal shedding of bacteria. The und...

  3. Inactivation of Bacillus cereus and Salmonella enterica serovar Typhimurium by aqueous ozone (O3): Modeling and Uv-Vis spectroscopic analysis

    Science.gov (United States)

    Ozone (O3) is a natural antimicrobial agent with potential applications in food industry. In this study, inactivation of Bacillus cereus and Salmonella enterica Typhimurium by aqueous ozone was evaluated. Ozone gas was generated using a domestic ozone generator with an output of 200 mg/hr (approx. 0...

  4. Effect of X-ray treatments on salmonella enterica and spoilage bacteria on skin-on chicken breast fillets and shell eggs

    Science.gov (United States)

    The objectives of this study were to determine the efficacy of X-ray irradiation on the inactivation of a 3- strain mixture of Salmonella enterica (S. Enteritidis E190-88, S. Typhimurium ATCC 14028, and S. Montevideo ATCC 8387) using an RS 2400 X-ray system on chicken breast fillets and shell eggs a...

  5. Characterization of extended-spectrum cephalosporin-resistant Salmonella enterica serovar Heidelberg isolated from food animals, retail meat, and humans in the United States 2009

    Science.gov (United States)

    Salmonella enterica is one of the most common causes of foodborne illness in the United States. Although salmonellosis is usually self-limiting, severe infections typically require antimicrobial treatment and ceftriaxone, an extended-spectrum cephalosporin, is commonly used in both adults and child...

  6. Development of a High-Throughput Multiplex PCR and Capillary Electrophoresis Technique for Serotype Determination of Salmonella Enterica Food Animal Isolates

    Science.gov (United States)

    Background: Previously, a multiplex PCR technique was developed to identify the top 30 human clinical serotypes of Salmonella enterica. To improve the speed, ease of use, utility and discriminatory ability of the technique, additional primers were added and the PCR product discrimination and analysi...

  7. Antimicrobial activities of red wine-based formulations containing plant extracts against Escherichia coli O157:H7 and Salmonella enterica serovar Hadar

    Science.gov (United States)

    We evaluated the bactericidal activities of red wine and red wine solutions containing a commercial olive extract called Hydrox-12, oregano oil, or a mixture of both in red wine against the following foodborne pathogens: Escherichia coli O157:H7, Salmonella enterica, Listeria monocytogenes, and Stap...

  8. Characterization of epidemic IncI1-Iγ plasmids harboring ambler class A and C genes in Escherichia coli and Salmonella enterica from animals and humans

    NARCIS (Netherlands)

    Smith, Hilde; Bossers, Alex; Harders, Frank; Wu, Guanghui; Woodford, Neil; Schwarz, Stefan; Guerra, Beatriz; Rodríguez, Irene; van Essen-Zandbergen, Alieda; Brouwer, Michael; Mevius, Dik

    2015-01-01

    The aim of the study was to identify the plasmid-encoded factors contributing to the emergence and spread of epidemic IncI1-Iγ plasmids obtained from Escherichia coli and Salmonella enterica isolates from animal and human reservoirs. For this, 251 IncI1-Iγ plasmids carrying various extended-spectrum

  9. Characterization of epidemic IncI1-Iγ plasmids harboring ambler class A and C genes in Escherichia coli and Salmonella enterica from animals and humans

    NARCIS (Netherlands)

    Smith, Hilde; Bossers, Alex; Harders, Frank; Wu, Guanghui; Woodford, Neil; Schwarz, Stefan; Guerra, Beatriz; Rodríguez, Irene; Essen-Zandbergen, Van Alieda; Brouwer, Michael; Mevius, Dik

    2015-01-01

    The aim of the study was to identify the plasmid-encoded factors contributing to the emergence and spread of epidemic IncI1-Iγ plasmids obtained from Escherichia coli and Salmonella enterica isolates from animal and human reservoirs. For this, 251 IncI1-Iγ plasmids carrying various extended-spect

  10. Detection of a blaSHV Extended-Spectrum β-Lactamase in Salmonella enterica Serovar Newport MDR-AmpC

    OpenAIRE

    Rankin, Shelley C.; Whichard, Jean M.; Joyce, Kevin; Stephens, Lynn; O'Shea, Kathleen; Aceto, Helen; Munro, Donald S.; Benson, Charles E.

    2005-01-01

    Salmonella enterica serovar Newport MDR-AmpC expressing TEM-1b and extended-spectrum β-lactamase SHV-12 was isolated from affected animals during an outbreak of salmonellosis that led to a 3-month closure of one of the largest equine hospitals in the United States.

  11. Highly expressed amino acid biosynthesis genes revealed by global gene expression analysis of Salmonella enterica serovar Enteritidis during growth in whole egg are not essential for this growth

    DEFF Research Database (Denmark)

    Jakočiūnė, Dzuiga; Herrero-Fresno, Ana; Jelsbak, Lotte;

    2016-01-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is the most common cause of egg borne salmonellosis in many parts of the world. This study analyzed gene expression of this bacterium during growth in whole egg, and whether highly expressed genes were essential for the growth. High quality...

  12. The evaluation of a PCR-based method for identification of Salmonella enterica serotypes from environmental samples and various food matrices.

    Science.gov (United States)

    Jean-Gilles Beaubrun, Junia; Cheng, Chorng-Ming; Chen, Kai-Shun; Ewing, Laura; Wang, Hua; Agpaoa, Maria C; Huang, Mei-Chiung J; Dickey, Erin; Du, Jamie M; Williams-Hill, Donna M; Hamilton, Brittany; Micallef, Shirley A; Rosenberg Goldstein, Rachel E; George, Ashish; Joseph, Sam W; Sapkota, Amy R; Jacobson, Andrew P; Tall, Ben D; Kothary, Mahendra H; Dudley, Kim; Hanes, Darcy E

    2012-09-01

    The most commonly used method for serotyping Salmonella spp. is based on the Kaufmann-White scheme, and is composed of serological reactions using antibodies to LPS agglutinins. The multiplex PCR used in this investigation was established by Kim et al. to serotype the 30 most common clinical Salmonella serotypes, as determined by CDC. The PCR assay consists of two five-plex reactions and a single two-plex PCR reaction, based on six genetic loci from Salmonella enterica serotype Typhimurium and four loci from S. enterica serotype Typhi. In this investigation, we further evaluated the method for serotyping Salmonella spp. using a reference collection, environmental samples collected from a Mid-Atlantic region tomato farm study, four food matrices spiked with different Salmonella serotypes and a proficiency test. The PCR assay was first evaluated using DNA isolated from pure cultures of isolates obtained from various clinical and environmental samples, and then DNA isolated from broth cultures of food matrices of "Red round" and Roma tomatoes, Romaine lettuce, green onions and Serrano peppers spiked with serotypes Newport, Typhimurium, Javiana and Saintpaul, respectively. The results showed that the PCR assay correctly serotyped Salmonella spp. from the clinical, environmental, spiked food matrices, and proficiency test samples. These findings are significant because the PCR assay was successful in the identification of Salmonella in the spiked samples in a broth culture containing other non-salmonella organism. This method may be a useful resource for the food safety community. PMID:22608224

  13. Influence of 5 major Salmonella pathogenicity islands on NK cell depletion in mice infected with Salmonella enterica serovar Enteritidis

    Directory of Open Access Journals (Sweden)

    Ondrackova Petra

    2010-03-01

    Full Text Available Abstract Background In this study we were interested in the colonisation and early immune response of Balb/C mice to infection with Salmonella Enteritidis and isogenic pathogenicity island free mutants. Results The virulence of S. Enteritidis for Balb/C mice was exclusively dependent on intact SPI-2. Infections with any of the mutants harbouring SPI-2 (including the mutant in which we left only SPI-2 but removed SPI-1, SPI-3, SPI-4 and SPI-5 resulted in fatalities, liver injures and NK cell depletion from the spleen. The infection was of minimal influence on counts of splenic CD4 CD8 T lymphocytes and γδ T-lymphocytes although a reduced ability of splenic lymphocytes to respond to non-specific mitogens indicated general immunosuppression in mice infected with SPI-2 positive S. Enteritidis mutants. Further investigations showed that NK cells were depleted also in blood but not in the caecal lamina propria. However, NK cell depletion was not directly associated with the presence of SPI-2 and was rather an indicator of virulence or avirulence of a particular mutant because the depletion was not observed in mice infected with other attenuated mutants such as lon and rfaL. Conclusions The virulence of S. Enteritidis for Balb/C mice is exclusively dependent on the presence of SPI-2 in its genome, and a major hallmark of the infection in terms of early changes in lymphocyte populations is the depletion of NK cells in spleen and blood. The decrease of NK cells in circulation can be used as a marker of attenuation of S. Enteritidis mutants for Balb/C mice.

  14. Microarray-based detection of Salmonella enterica serovar Typhimurium transposon mutants that cannot survive in macrophages and mice.

    Science.gov (United States)

    Chan, Kaman; Kim, Charles C; Falkow, Stanley

    2005-09-01

    DNA microarrays provide an opportunity to combine the principles of signature-tagged mutagenesis (STM) with microarray technology to identify potentially important bacterial virulence genes. The scope of DNA microarrays allows for less laborious screening on a much larger scale than possible by STM alone. We have adapted a microarray-based transposon tracking strategy for use with a Salmonella enterica serovar Typhimurium cDNA microarray in order to identify genes important for survival and replication in RAW 264.7 mouse macrophage-like cells or in the spleens of BALB/cJ mice. A 50,000-CFU transposon library of S. enterica serovar Typhimurium strain SL1344 was serially passaged in cultured macrophages or intraperitoneally inoculated into BALB/cJ mice. The bacterial genomic DNA was isolated and processed for analysis on the microarray. The novel application of this approach to identify mutants unable to survive in cultured cells resulted in the identification of components of Salmonella pathogenicity island 2 (SPI2), which is known to be critical for intracellular survival and replication. In addition, array results indicated that a number of SPI1-associated genes, currently not associated with intracellular survival, are negatively selected. However, of the SPI1-associated mutants individually tested for intracellular survival, only a sirA mutant exhibited reduced numbers relative to those of wild-type bacteria. Of the mutants unable to survive in mice, significant proportions are either components of the SPI2 pathogenicity island or involved in lipopolysaccharide synthesis. This observation is in agreement with results obtained in the original S. enterica serovar Typhimurium STM screen, illustrating the utility of this approach for the high-throughput identification of virulence factors important for survival in the host.

  15. Phage types of Salmonella enterica ssp. enterica serovar Typhimurium isolated from production animals and humans in Denmark

    DEFF Research Database (Denmark)

    Baggesen, Dorte Lau; Wegener, Henrik Caspar

    1994-01-01

    S. Typhimurium is one of the 2 most common salmonella serotypes causing human salmonellosis in Denmark. In order to illustrate the significance of different production animals as a source of infection, 1461 isolates were characterized by phage typing. The isolates originated from human patients a...

  16. Comparison of fecal culture and Danish Mix-ELISA for determination of Salmonella enterica subsp. enterica prevalence in growing swine.

    Science.gov (United States)

    Funk, J A; Harris, I T; Davies, P R

    2005-04-25

    In the USA, control of food-borne salmonellosis associated with meat consumption has been predominantly focused at slaughter and processing. It is expected that standards at slaughter and processing will become more stringent, creating pressure to reduce prevalence of Salmonella-positive food animals through on-farm interventions. The aim of this study was to compare traditional fecal culture and the Danish Mix-ELISA (DME) for determination of Salmonella prevalence pre-harvest in swine. In Trial 1, five cohorts of individually identified pigs were longitudinally sampled during the growing period to compare the kinetics of prevalence as estimated by fecal culture and the DME. In Trial 2, the correlation between fecal prevalence and seroprevalence was estimated pre-marketing in 49 groups of pigs. In Trial 1, fecal prevalence and seroprevalence showed similar kinetics, with a tendency of a higher OD% cut-off to more closely approximate fecal prevalence. In Trial 2, correlations between fecal culture and the DME were 0.40, 0.36, 0.43, and 0.43 (p or =10, 20, 30, and 40, respectively. Based on these results, a higher OD% cut-off would be recommended if more approximate estimation of fecal prevalence is desired and longitudinal sampling would be suggested for evaluating the impact of on-farm interventions for Salmonella reduction whether utilizing fecal culture or the DME. Further evaluation of the impact of Salmonella serovar present on farms on seroprevalence and the relationship of on-farm seroprevalence with food safety risk are needed prior to utilizing the DME for pre-harvest Salmonella diagnostics in the US swine herd. PMID:15795083

  17. Infection of Murine Macrophages by Salmonella enterica Serovar Heidelberg Blocks Murine Norovirus Infectivity and Virus-induced Apoptosis.

    Directory of Open Access Journals (Sweden)

    Sudhakar S Agnihothram

    Full Text Available Gastroenteritis caused by bacterial and viral pathogens constitutes a major public health threat in the United States accounting for 35% of hospitalizations. In particular, Salmonella enterica and noroviruses cause the majority of gastroenteritis infections, with emergence of sporadic outbreaks and incidence of increased infections. Although mechanisms underlying infections by these pathogens have been individually studied, little is known about the mechanisms regulating co-infection by these pathogens. In this study, we utilized RAW 264.7 murine macrophage cells to investigate the mechanisms governing co-infection with S. enterica serovar Heidelberg and murine norovirus (MNV. We demonstrate that infection of RAW 264.7 cells with S. enterica reduces the replication of MNV, in part by blocking virus entry early in the virus life cycle, and inducing antiviral cytokines later in the infection cycle. In particular, bacterial infection prior to, or during MNV infection affected virus entry, whereas MNV entry remained unaltered when the virus infection preceded bacterial invasion. This block in virus entry resulted in reduced virus replication, with the highest impact on replication observed during conditions of co-infection. In contrast, bacterial replication showed a threefold increase in MNV-infected cells, despite the presence of antibiotic in the medium. Most importantly, we present evidence that the infection of MNV-infected macrophages by S. enterica blocked MNV-induced apoptosis, despite allowing efficient virus replication. This apoptosis blockade was evidenced by reduction in DNA fragmentation and absence of poly-ADP ribose polymerase (PARP, caspase 3 and caspase 9 cleavage events. Our study suggests a novel mechanism of pathogenesis whereby initial co-infection with these pathogens could result in prolonged infection by either of these pathogens or both together.

  18. Mass spectrometry-based quantitative proteomic analysis of Salmonella enterica serovar Enteritidis protein expression upon exposure to hydrogen peroxide

    Directory of Open Access Journals (Sweden)

    Su Jing

    2010-06-01

    Full Text Available Abstract Background Salmonella enterica, a common food-borne bacterial pathogen, is believed to change its protein expression profile in the presence of different environmental stress such as that caused by the exposure to hydrogen peroxide (H2O2, which can be generated by phagocytes during infection and represents an important antibacterial mechanism of host cells. Among Salmonella proteins, the effectors of Salmonella pathogenicity island 1 and 2 (SPI-1 and SPI-2 are of particular interest since they are expressed during host infection in vivo and are important for invasion of epithelial cells and for replication in organs during systemic infection, respectively. However, the expression profiles of these proteins upon exposure to H2O2 or to host cells in vivo during the established phase of systemic infection have not been extensively studied. Results Using stable isotope labeling coupled with mass spectrometry, we performed quantitative proteomic analysis of Salmonella enterica serovar Enteritidis and identified 76 proteins whose expression is modulated upon exposure to H2O2. SPI-1 effector SipC was expressed about 3-fold higher and SopB was expressed approximately 2-fold lower in the presence of H2O2, while no significant change in the expression of another SPI-1 protein SipA was observed. The relative abundance of SipA, SipC, and SopB was confirmed by Western analyses, validating the accuracy and reproducibility of our approach for quantitative analysis of protein expression. Furthermore, immuno-detection showed substantial expression of SipA and SipC but not SopB in the late phase of infection in macrophages and in the spleen of infected mice. Conclusions We have identified Salmonella proteins whose expression is modulated in the presence of H2O2. Our results also provide the first direct evidence that SipC is highly expressed in the spleen at late stage of salmonellosis in vivo. These results suggest a possible role of SipC and other

  19. 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

  20. Horizontal gene transfer of a ColV plasmid has resulted in a dominant avian clonal type of Salmonella enterica serovar Kentucky.

    Directory of Open Access Journals (Sweden)

    Timothy J Johnson

    Full Text Available Salmonella enterica continues to be a significant cause of foodborne gastrointestinal illness in humans. A wide variety of Salmonella serovars have been isolated from production birds and from retail poultry meat. Recently, though, S. enterica subsp. enterica serovar Kentucky has emerged as one of the prominent Salmonella serovars isolated from broiler chickens. Recent work suggests that its emergence apparently coincides with its acquisition of a ColV virulence plasmid. In the present study, we examined 902 Salmonella isolates belonging to 59 different serovars for the presence of this plasmid. Of the serovars examined, the ColV plasmid was found only among isolates belonging to the serovars Kentucky (72.9%, Typhimurium (15.0% and Heidelberg (1.7%. We demonstrated that a single PFGE clonal type of S. Kentucky harbors this plasmid, and acquisition of this plasmid by S. Kentucky significantly increased its ability to colonize the chicken cecum and cause extraintestinal disease. Comparison of the completed sequences of three ColV plasmids from S. Kentucky isolated from different geographical locales, timepoints and sources revealed a nearly identical genetic structure with few single nucleotide changes or insertions/deletions. Overall, it appears that the ColV plasmid was recently acquired by a single clonal type S. Kentucky and confers to its host enhanced colonization and fitness capabilities. Thus, the potential for horizontal gene transfer of virulence and fitness factors to Salmonella from other enteric bacteria exists in poultry, representing a potential human health hazard.

  1. A multiplex single nucleotide polymorphism typing assay for detecting mutations that result in decreased fluoroquinolone susceptibility in Salmonella enterica serovars Typhi and Paratyphi A.

    LENUS (Irish Health Repository)

    Song, Yajun

    2010-08-01

    OBJECTIVES: Decreased susceptibility to fluoroquinolones has become a major problem for the successful therapy of human infections caused by Salmonella enterica, especially the life-threatening typhoid and paratyphoid fevers. METHODS: By using Luminex xTAG beads, we developed a rapid, reliable and cost-effective multiplexed genotyping assay for simultaneously detecting 11 mutations in gyrA, gyrB and parE of S. enterica serovars Typhi and Paratyphi A that result in nalidixic acid resistance (Nal(R)) and\\/or decreased susceptibility to fluoroquinolones. RESULTS: This assay yielded unambiguous single nucleotide polymorphism calls on extracted DNA from 292 isolates of Salmonella Typhi (Nal(R) = 223 and Nal(S) = 69) and 106 isolates of Salmonella Paratyphi A (Nal(R) = 24 and Nal(S) = 82). All of the 247 Nal(R) Salmonella Typhi and Salmonella Paratyphi A isolates were found to harbour at least one of the target mutations, with GyrA Phe-83 as the most common one (143\\/223 for Salmonella Typhi and 18\\/24 for Salmonella Paratyphi A). We also identified three GyrB mutations in eight Nal(S) Salmonella Typhi isolates (six for GyrB Phe-464, one for GyrB Leu-465 and one for GyrB Asp-466), and mutations GyrB Phe-464 and GyrB Asp-466 seem to be related to the decreased ciprofloxacin susceptibility phenotype in Salmonella Typhi. This assay can also be used directly on boiled single colonies. CONCLUSIONS: The assay presented here would be useful for clinical and reference laboratories to rapidly screen quinolone-resistant isolates of Salmonella Typhi and Salmonella Paratyphi A, and decipher the underlying genetic changes for epidemiological purposes.

  2. Effect of chemical sanitizers on Salmonella enterica serovar Poona on the surface of cantaloupe and pathogen contamination of internal tissues as a function of cutting procedure.

    Science.gov (United States)

    Vadlamudi, Sasi; Taylor, T Matthew; Blankenburg, Charles; Castillo, Alejandro

    2012-10-01

    Survival of Salmonella enterica subsp. enterica serovar Poona on surface and stem scar portions of inoculated cantaloupe following sanitizer application, transfer of pathogen from the rind to the flesh during cutting, and growth of Salmonella Poona on cantaloupe cubes over 15 days of refrigerated storage were investigated. Cantaloupes inoculated with a rifampin-resistant strain of Salmonella Poona (10(7) CFU/ml) for 3 min and dried for 12 h were washed with chlorine (200 mg free chlorine per liter, 3 min), lactic acid (2%, 2 min), or ozone (30 mg/liter, 5 min). Fresh-cut cantaloupe cubes were prepared by (i) cutting the cantaloupe and then removing the rind or by (ii) peeling the rind and then cutting the flesh into pieces. The numbers of Salmonella bacteria recovered were higher in the stem scar portion (6.3 ± 0.3 log CFU/cm(2)) than the surface (4.8 ± 0.2 log CFU/cm(2)). Surface treatment with tap water or chlorine did not reduce Salmonella numbers, while treatment with lactic acid or ozone reduced Salmonella by 2.5 or 2.3 log CFU/cm(2), respectively. The use of lactic acid to sanitize the cantaloupes resulted in less Salmonella transfer to flesh during cutting; Salmonella numbers decreased to below detectable levels over 9 days of refrigerated (4°C) storage. Cutting cantaloupes after peeling the rind was more effective at reducing transfer of Salmonella to the internal tissue than cutting of cantaloupes prior to rind removal. These data suggest that treatment of cantaloupe rinds with lactic acid or ozone may be effective at reducing Salmonella numbers, while lactic acid application resulted in reduction of Salmonella transfer to cantaloupe flesh. PMID:23043824

  3. Antimicrobial susceptibility and virulence characteristics of Salmonella enterica Typhimurium isolates from healthy and diseased pigs in Korea.

    Science.gov (United States)

    Tamang, Migma Dorji; Gurung, Mamata; Nam, Hyang-Mi; Moon, Dong Chan; Jang, Geum-Chan; Jung, Suk-Chan; Lim, Suk-Kyung

    2014-09-01

    This study compared the antimicrobial susceptibility and prevalence of virulence genes in Salmonella enterica Typhimurium isolated from healthy and diseased pigs in Korea. A total of 456 Salmonella Typhimurium isolated from healthy (n = 238) and diseased (n = 218) pigs between 1998 and 2011 were investigated. In total, 93.4% of the Salmonella Typhimurium isolates were resistant to at least one antimicrobial agent tested. The isolates were most often resistant to tetracycline (85.7%), followed by streptomycin (83.6%), nalidixic acid (67.3%), ampicillin (49.3%), chloramphenicol (42.8%), and gentamicin (37.1%). Moreover, multidrug resistance phenotype and resistance to ampicillin, florfenicol, gentamicin, nalidixic acid, neomycin, streptomycin, and tetracycline were significantly higher (P invA, spiA, msgA, sipB, prgH, spaN, tolC, lpfC, sifA, sitC, and sopB virulence genes. The prevalence of orgA, pagC, and iroN were 50.2, 74.1, and 91.0%, respectively, whereas isolates carrying cdtB (1.5%), pefA (7.0%), and spvB (14.9%) were identified much less frequently. Furthermore, the prevalence of invA, lpfC, orgA, pagC, and iroN was significantly higher (P < 0.01) among the isolates from the diseased pigs than in isolates from the healthy pigs. Our results demonstrated that, among diseased pigs, there was significantly higher resistance to some antimicrobials and greater prevalence of some virulence genes than in healthy pigs, indicating the role these factors play in pathogenesis. Multidrug-resistant Salmonella isolates that carry virulence-associated genes are potentially more dangerous and constitute a public health concern. Thus, continuous surveillance of antimicrobial resistance and virulence characteristics in Salmonella is essential.

  4. 15-Deoxy-Δ12,14-prostaglandin J2 inhibits macrophage colonization by Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Buckner, Michelle M C; Antunes, L Caetano M; Gill, Navkiran; Russell, Shannon L; Shames, Stephanie R; Finlay, B Brett

    2013-01-01

    15-deoxy-Δ(12,14)-prostaglandin J2 (15d-PGJ2) is an anti-inflammatory downstream product of the cyclooxygenase enzymes. It has been implicated to play a protective role in a variety of inflammatory mediated diseases, including rheumatoid arthritis, neural damage, and myocardial infarctions. Here we show that 15d-PGJ2 also plays a role in Salmonella infection. Salmonella enterica Typhimurium is a Gram-negative facultative intracellular pathogen that is able to survive and replicate inside phagocytic immune cells, allowing for bacterial dissemination to systemic sites. Salmonella species cause a wide range of morbidity and mortality due to gastroenteritis and typhoid fever. Previously we have shown that in mouse models of typhoid fever, Salmonella infection causes a major perturbation in the prostaglandin pathway. Specifically, we saw that 15d-PGJ2 production was significantly increased in both liver and feces. In this work we show that 15d-PGJ2 production is also significantly increased in macrophages infected with Salmonella. Furthermore, we show that the addition of 15d-PGJ2 to Salmonella infected RAW264.7, J774, and bone marrow derived macrophages is sufficient to significantly reduce bacterial colonization. We also show evidence that 15d-PGJ2 is reducing bacterial uptake by macrophages. 15d-PGJ2 reduces the inflammatory response of these infected macrophages, as evidenced by a reduction in the production of cytokines and reactive nitrogen species. The inflammatory response of the macrophage is important for full Salmonella virulence, as it can give the bacteria cues for virulence. The reduction in bacterial colonization is independent of the expression of Salmonella virulence genes SPI1 and SPI2, and is independent of the 15d-PGJ2 ligand PPAR-γ. 15d-PGJ2 also causes an increase in ERK1/2 phosphorylation in infected macrophages. In conclusion, we show here that 15d-PGJ2 mediates the outcome of bacterial infection, a previously unidentified role for this

  5. 15-Deoxy-Δ12,14-prostaglandin J2 inhibits macrophage colonization by Salmonella enterica serovar Typhimurium.

    Directory of Open Access Journals (Sweden)

    Michelle M C Buckner

    Full Text Available 15-deoxy-Δ(12,14-prostaglandin J2 (15d-PGJ2 is an anti-inflammatory downstream product of the cyclooxygenase enzymes. It has been implicated to play a protective role in a variety of inflammatory mediated diseases, including rheumatoid arthritis, neural damage, and myocardial infarctions. Here we show that 15d-PGJ2 also plays a role in Salmonella infection. Salmonella enterica Typhimurium is a Gram-negative facultative intracellular pathogen that is able to survive and replicate inside phagocytic immune cells, allowing for bacterial dissemination to systemic sites. Salmonella species cause a wide range of morbidity and mortality due to gastroenteritis and typhoid fever. Previously we have shown that in mouse models of typhoid fever, Salmonella infection causes a major perturbation in the prostaglandin pathway. Specifically, we saw that 15d-PGJ2 production was significantly increased in both liver and feces. In this work we show that 15d-PGJ2 production is also significantly increased in macrophages infected with Salmonella. Furthermore, we show that the addition of 15d-PGJ2 to Salmonella infected RAW264.7, J774, and bone marrow derived macrophages is sufficient to significantly reduce bacterial colonization. We also show evidence that 15d-PGJ2 is reducing bacterial uptake by macrophages. 15d-PGJ2 reduces the inflammatory response of these infected macrophages, as evidenced by a reduction in the production of cytokines and reactive nitrogen species. The inflammatory response of the macrophage is important for full Salmonella virulence, as it can give the bacteria cues for virulence. The reduction in bacterial colonization is independent of the expression of Salmonella virulence genes SPI1 and SPI2, and is independent of the 15d-PGJ2 ligand PPAR-γ. 15d-PGJ2 also causes an increase in ERK1/2 phosphorylation in infected macrophages. In conclusion, we show here that 15d-PGJ2 mediates the outcome of bacterial infection, a previously unidentified

  6. Attenuated Salmonella typhimurium SV4089 as a potential carrier of oral DNA vaccine in chickens.

    Science.gov (United States)

    Jazayeri, Seyed Davoud; Ideris, Aini; Zakaria, Zunita; Omar, Abdul Rahman

    2012-01-01

    Attenuated Salmonella has been used as a carrier for DNA vaccine. However, in vitro and in vivo studies on the bacteria following transfection of plasmid DNA were poorly studied. In this paper, eukaryotic expression plasmids encoding avian influenza virus (AIV) subtype H5N1 genes, pcDNA3.1/HA, NA, and NP, were transfected into an attenuated Salmonella enteric typhimurium SV4089. In vitro stability of the transfected plasmids into Salmonella were over 90% after 100 generations. The attenuated Salmonella were able to invade MCF-7 (1.2%) and MCF-10A (0.5%) human breast cancer cells. Newly hatched specific-pathogen-free (SPF) chicks were inoculated once by oral gavage with 10(9) colony-forming unit (CFU) of the attenuated Salmonella. No abnormal clinical signs or deaths were recorded after inoculation. Viable bacteria were detected 3 days after inoculation by plating from spleen, liver, and cecum. Fluorescent in situ hybridization (FISH) and polymerase chain reaction (PCR) were carried out for confirmation. Salmonella was not detected in blood cultures although serum antibody immune responses to Salmonella O antiserum group D1 factor 1, 9, and 12 antigens were observed in all the inoculated chickens after 7 days up to 35 days. Our results showed that live attenuated S. typhimurium SV4089 harboring pcDNA3.1/HA, NA, and NP may provide a unique alternative as a carrier for DNA oral vaccine in chickens.

  7. Highly expressed amino acid biosynthesis genes revealed by global gene expression analysis of Salmonella enterica serovar Enteritidis during growth in whole egg are not essential for this growth.

    Science.gov (United States)

    Jakočiūnė, Džiuginta; Herrero-Fresno, Ana; Jelsbak, Lotte; Olsen, John Elmerdahl

    2016-05-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is the most common cause of egg borne salmonellosis in many parts of the world. This study analyzed gene expression of this bacterium during growth in whole egg, and whether highly expressed genes were essential for the growth. High quality RNA was extracted from S. Enteritidis using a modified RNA-extraction protocol. Global gene expression during growth in whole egg was compared to growth in LB-medium using DNA array method. Twenty-six genes were significantly upregulated during growth in egg; these belonged to amino acid biosynthesis, di/oligopeptide transport system, biotin synthesis, ferrous iron transport system, and type III secretion system. Significant downregulation of 15 genes related to formate hydrogenlyase (FHL) and trehalose metabolism was observed. The results suggested that S. Enteritidis is starved for amino-acids, biotin and iron when growing in egg. However, site specific mutation of amino acid biosynthesis genes asnA (17.3 fold upregulated), asnB (18.6 fold upregulated), asnA/asnB and, serA (12.0 fold upregulated) and gdhA (3.7 fold upregulated), did not result in growth attenuation, suggesting that biosynthesis using the enzymes encoded from these genes may represent the first choice for S. Enteritidis when growing in egg, but when absent, the bacterium could use alternative ways to obtain the amino acids.

  8. Split Marketing as a Risk Factor for Salmonella Enterica Infection in Swine

    Science.gov (United States)

    On-farm reduction of Salmonella prevalence in pigs requires the identification of risk factors to direct interventions. This study was designed to determine if split marketing of finishing pigs constitutes a risk factor for Salmonella infections, by comparing Salmonella prevalence in the first group...

  9. Evaluation of Salmonella enterica serovar Enteritidis pathogenicity island-1 proteins as vaccine candidates against S. Enteritidis challenge in chickens.

    Science.gov (United States)

    Desin, Taseen S; Wisner, Amanda L S; Lam, Po-King S; Berberov, Emil; Mickael, Claudia S; Potter, Andrew A; Köster, Wolfgang

    2011-03-24

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major cause of gastrointestinal disease in humans worldwide, which mainly results from the consumption of contaminated poultry meat and eggs. Vaccination of chickens is an important strategy to lower the prevalence of Salmonella in poultry flocks. The S. Enteritidis type 3 secretion system (T3SS) encoded on Salmonella pathogenicity island-1 (SPI-1) is an important virulence factor that plays a role in invasion and systemic spread in chickens. In this manuscript, we evaluated the efficacy of SPI-1 proteins as vaccine candidates for protection against S. Enteritidis oral challenge. Our results demonstrate for the first time that SPI-1 T3SS proteins elicit antigen specific IgG antibody responses in chickens. In one study we show that vaccination with the aforementioned proteins reduces the levels of S. Enteritidis in the liver, but not in the spleen and cecal contents of chickens. However, a second study shows that vaccination of hens with SPI-1 proteins using a seeder model of infection does not affect the levels of S. Enteritidis in the cecal contents or internal organs of progeny obtained from these hens. Hence, the SPI-1 proteins, in conjunction with other proteins, may form important components of subunit vaccines used for protection against colonization by S. Enteritidis in poultry.

  10. Discovery of Novel Secreted Virulence Factors from Salmonella enterica Serovar Typhimurium by Proteomic Analysis of Culture Supernatants

    Energy Technology Data Exchange (ETDEWEB)

    Niemann, George; Brown, Roslyn N.; Gustin, Jean K.; Stufkens, Afke; Shaikh-Kidwai, Afshan S.; Li, Jie; McDermott, Jason E.; Brewer, Heather M.; Schepmoes, Athena A.; Smith, Richard D.; Adkins, Joshua N.; Heffron, Fred

    2011-01-01

    The intracellular pathogen Salmonella enterica serovar Typhimurium is a leading cause of acute gastroenteritis in the world. This pathogen has two type-III secretion systems (TTSS) necessary for virulence that are encoded in Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2) and are expressed during extracellular or intracellular infectious states, respectively, to deliver virulence factors (effectors) to the host cell cytoplasm. While many have been identified and at least partially characterized, the full repertoire of effectors has not been catalogued. In this mass spectrometry-based proteomics study, we identified effector proteins secreted under minimal acidic medium growth conditions that induced the SPI-2 TTSS and its effectors, and compared the secretome from the parent strain to the secretome from strains missing either essential (SsaK) or regulatory components (SsaL) of the SPI-2 secretion apparatus. We identified 75% of the known TTSS effector repertoire. Excluding translocon components, 95% of the known effectors were biased for identification in the ssaL mutant background, which demonstrated that SsaL regulates SPI-2 type III secretion. To confirm secretion to animal cells, we made translational fusions of several of the best candidates to the calmodulin-dependent adenylate cyclase of Bordetella pertussis and assayed cAMP levels of infected J774 macrophage-like cells. From these infected cells we identified six new TTSS effectors and two others that are secreted independent of TTSS. Our results substantiate reports of additional secretion systems encoded by Salmonella other than TTSS.

  11. Antimicrobial resistance in Salmonella enterica serovar typhi and paratyphi in South Asia-current status, issues and prospects.

    Science.gov (United States)

    Akhtar, Saeed; Sarker, Mahfuzur R; Jabeen, Kausar; Sattar, Ahsan; Qamar, Aftab; Fasih, Naima

    2015-01-01

    The human race owes a debt of gratitude to antimicrobial agents, penicillin and its successors that have saved people from tremendous pain and suffering in the last several decades. Unfortunately, this consideration is no more true, as millions of people are prone to the challenging threat of emergence of antimicrobial resistance worldwide and the menace is more distressing in developing countries. Comparable with other bacterial species, Salmonella enterica serovar Typhi (S. typhi) and Paratyphi (S. paratyphi) have been evolving multidrug resistance (MDR) against a wide array of antibiotics, including chloramphenicol, ampicillin and co-trimoxazole, and globally affecting 21 million people with 220,000 deaths each year. S. typhi and S. paratyphi infections are also endemic in South Asia and a series of antibiotics used to treat these infections, have been losing efficacy against enteric fever. Currently, quinolones are regarded as a choice to treat MDR Salmonella in these regions. Travel-related cases of enteric fever, especially from South Asian countries are the harbinger of the magnitude of MDR Salmonella in that region. Conclusively, the MDR will continue to grow and the available antimicrobial agents would become obsolete. Therefore, a radical and aggressive approach in terms of rational use of antibiotics during treating infections is essentially needed. PMID:24645636

  12. Atmospheric cold plasma inactivation of Escherichia coli, Salmonella enterica serovar Typhimurium and Listeria monocytogenes inoculated on fresh produce.

    Science.gov (United States)

    Ziuzina, D; Patil, S; Cullen, P J; Keener, K M; Bourke, P

    2014-09-01

    Atmospheric cold plasma (ACP) represents a potential alternative to traditional methods for non-thermal decontamination of foods. In this study, the antimicrobial efficacy of a novel dielectric barrier discharge ACP device against Escherichia coli, Salmonella enterica Typhimurium and Listeria monocytogenes inoculated on cherry tomatoes and strawberries, was examined. Bacteria were spot inoculated on the produce surface, air dried and sealed inside a rigid polypropylene container. Samples were indirectly exposed (i.e. placed outside plasma discharge) to a high voltage (70 kVRMS) air ACP and subsequently stored at room temperature for 24 h. ACP treatment for 10, 60 and 120 s resulted in reduction of Salmonella, E. coli and L. monocytogenes populations on tomato to undetectable levels from initial populations of 3.1, 6.3, and 6.7 log10 CFU/sample, respectively. However, an extended ACP treatment time was necessary to reduce bacterial populations attached on the more complex surface of strawberries. Treatment time for 300 s resulted in reduction of E. coli, Salmonella and L. monocytogenes populations by 3.5, 3.8 and 4.2 log10 CFU/sample, respectively, and also effectively reduced the background microflora of tomatoes.

  13. Evaluation of Salmonella enterica serovar Enteritidis pathogenicity island-1 proteins as vaccine candidates against S. Enteritidis challenge in chickens.

    Science.gov (United States)

    Desin, Taseen S; Wisner, Amanda L S; Lam, Po-King S; Berberov, Emil; Mickael, Claudia S; Potter, Andrew A; Köster, Wolfgang

    2011-03-24

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major cause of gastrointestinal disease in humans worldwide, which mainly results from the consumption of contaminated poultry meat and eggs. Vaccination of chickens is an important strategy to lower the prevalence of Salmonella in poultry flocks. The S. Enteritidis type 3 secretion system (T3SS) encoded on Salmonella pathogenicity island-1 (SPI-1) is an important virulence factor that plays a role in invasion and systemic spread in chickens. In this manuscript, we evaluated the efficacy of SPI-1 proteins as vaccine candidates for protection against S. Enteritidis oral challenge. Our results demonstrate for the first time that SPI-1 T3SS proteins elicit antigen specific IgG antibody responses in chickens. In one study we show that vaccination with the aforementioned proteins reduces the levels of S. Enteritidis in the liver, but not in the spleen and cecal contents of chickens. However, a second study shows that vaccination of hens with SPI-1 proteins using a seeder model of infection does not affect the levels of S. Enteritidis in the cecal contents or internal organs of progeny obtained from these hens. Hence, the SPI-1 proteins, in conjunction with other proteins, may form important components of subunit vaccines used for protection against colonization by S. Enteritidis in poultry. PMID:20888713

  14. Analysis of plasmid-mediated quinolone and oxyimino-cephalosporin resistance mechanisms in Uruguayan Salmonella enterica isolates from 2011-2013.

    Science.gov (United States)

    Cordeiro, Nicolás F; Nabón, Adriana; García-Fulgueiras, Virginia; Álvez, Marcelo; Sirok, Alfredo; Camou, Teresa; Vignoli, Rafael

    2016-09-01

    This study characterised the mechanisms of fluoroquinolone and oxyimino-cephalosporin resistance in human Salmonella enterica isolates in Uruguay. Salmonella enterica isolates were collected from 2011-2013 and were selected based on non-susceptibility to ciprofloxacin and/or oxyimino-cephalosporins. The disk diffusion assay was performed for various antibiotics, and the ciprofloxacin minimum inhibitory concentration (MIC) was determined following CLSI guidelines. Genetic relatedness was determined following PulseNet protocols. Extended-spectrum β-lactamases, ampC alleles and plasmid-mediated quinolone resistance were characterised by PCR and sequencing. Plasmid analyses were carried out by conjugation or transformation assays, and plasmid-encoded genes were identified by PCR. Mutations in the quinolone resistance-determining region of gyrases were sought by PCR and sequencing. Among 579 isolates, 105 (18.4%) ciprofloxacin-non-susceptible (CIP-NS) isolates, 9 (1.6%) oxyimino-cephalosporin-resistant isolates and 2 (0.3%) isolates resistant to both antibiotic families were detected. Thirteen isolates carried qnrB alleles (twelve qnrB19 and one qnrB2), four carried blaCTX-M-8, two blaCTX-M-14, two blaSHV-2 and three blaCMY-2-like genes. No correlation was found between mutations in gyrases and ciprofloxacin MICs. Several co-circulating clones of S. enterica ssp. enterica serovar Typhimurium were detected; conversely, S. enterica ssp. enterica serovar Enteritidis corresponded mainly to a single circulating clone. Nine (75%) of twelve of CIP-NS extraintestinal isolates shared the same pulsotype with intestinal isolates. During the study period, the frequency of CIP-NS isolates increased, albeit with ciprofloxacin MICs of 0.125-0.5mg/L. Detection of the same quinolone-resistant clones recovered both from intestinal and extraintestinal samples highlights the significance of epidemiological surveillance of antibiotic susceptibility for every human Salmonella isolate. PMID

  15. Optimization of rapid Salmonella enterica detection in liquid whole eggs by SYBR green I-based real-time reverse transcriptase-polymerase chain reaction.

    Science.gov (United States)

    Techathuvanan, Chayapa; D'Souza, Doris Helen

    2011-04-01

    Eggs and egg products have a high risk of Salmonella enterica serovar Enteritidis contamination leading to gastroenteritis outbreaks in humans. Thus, a rapid screening tool for viable Salmonella Enteritidis cells in the egg industry is needed. Our objective was to rapidly and sensitively detect viable Salmonella Enteritidis from spiked liquid whole eggs (LWEs) within 24 h using SYBR green I-based real-time reverse transcriptase-polymerase chain reaction (PCR) targeting the Salmonella specific invA gene along with an internal amplification control in a Bio-Rad iCycler. LWE was inoculated with Salmonella Enteritidis and mixed with tetrathionate broth, and 100 μL of serially diluted portions in phosphate-buffered saline was plated on Xylose Lysine Tergitol 4 agar or 5 mL were used for RNA extraction by the TRIzol method immediately or after enrichment of 6, 12, or 16 h at 37 °C. The real-time reverse transcriptase-PCR assay was carried out using previously described Salmonella invA gene primers. Melt temperature analysis of the PCR product was included to determine specific invA amplification. Without enrichment, the assay detection limit was 10(7) colony forming units (CFU)/25 mL LWE. After enrichment for 6 and 12 h, Salmonella Enteritidis could be detected from LWE up to 10(4) and 10(2) CFU/25 mL, respectively. Improved Salmonella Enteritidis detection up to 10(0) CFU/25 mL was obtained after 16-h enrichment. Even with 16-h enrichment, the results could be still be obtained within 24 h, which is much faster than by traditional cultural detection that takes several days. Therefore, this assay appears suitable for routine detection of Salmonella enterica contamination by the egg industry to help prevent the transmission of egg-associated Salmonella outbreaks and timely recall of contaminated products.

  16. Attenuation of 10 MeV electron beam energy to achieve low doses does not affect Salmonella spp. inactivation kinetics

    Science.gov (United States)

    Hieke, Anne-Sophie Charlotte; Pillai, Suresh D.

    2015-05-01

    The effect of attenuating the energy of a 10 MeV electron beam on Salmonella inactivation kinetics was investigated. No statistically significant differences were observed between the D10 values of either Salmonella 4,[5],12:i:- or a Salmonella cocktail (S. 4,[5],12:i:-, Salmonella Heidelberg, Salmonella Newport, Salmonella Typhimurium, Salmonella) when irradiated with either a non-attenuated 10 MeV eBeam or an attenuated 10 MeV eBeam (~2.9±0.22 MeV). The results show that attenuating the energy of a 10 MeV eBeam to achieve low doses does not affect the inactivation kinetics of Salmonella spp. when compared to direct 10 MeV eBeam irradiation.

  17. Strain-Specific Survival of Salmonella enterica in Peanut Oil, Peanut Shell, and Chia Seeds.

    Science.gov (United States)

    Fong, Karen; Wang, Siyun

    2016-03-01

    In North America, outbreaks of Salmonella have been linked to low-water activity (aw) foods, such as nuts and seeds. These outbreaks have implicated an assortment of Salmonella serotypes. Some Salmonella serotypes (e.g., Enteritidis and Typhimurium) cause high proportions of salmonellosis. Nevertheless, there has recently been an emergence of uncommon Salmonella serotypes and strains (e.g., Tennessee, Hartford, and Thompson) in low-aw foods. The aim of this study was to evaluate the survival characteristics of Salmonella serotypes Enteritidis, Typhimurium, Tennessee, Hartford, and Thompson in three low-aw food ingredients with varying aw: peanut oil (aw = 0.521 ± 0.003), peanut shell (aw = 0.321 ± 0.20), and chia seeds (aw = 0.585 ± 0.003). The survival of individual Salmonella strains on each food matrix was monitored for a maximum of 150 days by spreading the bacterial cells onto Luria-Bertani and/or xylose lysine deoxycholate agar. Overall, Salmonella survived for the longest periods of time in peanut oil (96 ± 8 days), followed by chia seeds (94 ± 46 days). The survival period was substantially reduced on the surface of peanut shell (42 ± 49 h), although PCR after 70 days of incubation revealed the presence of Salmonella cells. In addition, Salmonella exhibited a strain-specific response in the three low-aw foods tested. Salmonella Hartford was identified as highly persistent in all low-aw food matrices, whereas Salmonella Typhimurium was the least persistent. The current research emphasizes the adaptable nature of Salmonella to low-aw food ingredients. This may pose additional problems owing to the downstream production of various end products. Additionally, unique survival characteristics among Salmonella strains highlight the need for tailored mitigation strategies regarding high-risk Salmonella strains in the food industry. PMID:26939645

  18. Strain-Specific Survival of Salmonella enterica in Peanut Oil, Peanut Shell, and Chia Seeds.

    Science.gov (United States)

    Fong, Karen; Wang, Siyun

    2016-03-01

    In North America, outbreaks of Salmonella have been linked to low-water activity (aw) foods, such as nuts and seeds. These outbreaks have implicated an assortment of Salmonella serotypes. Some Salmonella serotypes (e.g., Enteritidis and Typhimurium) cause high proportions of salmonellosis. Nevertheless, there has recently been an emergence of uncommon Salmonella serotypes and strains (e.g., Tennessee, Hartford, and Thompson) in low-aw foods. The aim of this study was to evaluate the survival characteristics of Salmonella serotypes Enteritidis, Typhimurium, Tennessee, Hartford, and Thompson in three low-aw food ingredients with varying aw: peanut oil (aw = 0.521 ± 0.003), peanut shell (aw = 0.321 ± 0.20), and chia seeds (aw = 0.585 ± 0.003). The survival of individual Salmonella strains on each food matrix was monitored for a maximum of 150 days by spreading the bacterial cells onto Luria-Bertani and/or xylose lysine deoxycholate agar. Overall, Salmonella survived for the longest periods of time in peanut oil (96 ± 8 days), followed by chia seeds (94 ± 46 days). The survival period was substantially reduced on the surface of peanut shell (42 ± 49 h), although PCR after 70 days of incubation revealed the presence of Salmonella cells. In addition, Salmonella exhibited a strain-specific response in the three low-aw foods tested. Salmonella Hartford was identified as highly persistent in all low-aw food matrices, whereas Salmonella Typhimurium was the least persistent. The current research emphasizes the adaptable nature of Salmonella to low-aw food ingredients. This may pose additional problems owing to the downstream production of various end products. Additionally, unique survival characteristics among Salmonella strains highlight the need for tailored mitigation strategies regarding high-risk Salmonella strains in the food industry.

  19. Identificazione e caratterizzazione dei determinanti genetici di antibiotico-resistenza in ceppi di Salmonella enterica di origine animale

    Directory of Open Access Journals (Sweden)

    Cristina Pezzella

    2004-12-01

    Full Text Available Tetracyclines are broad-spectrum agents, exhibiting activity against a wide range of gram-positive and gram-negative bacteria and are currently used for therapy and prophylaxis of human infections and for the prevention and control of bacterial infections in veterinary medicine. Streptomycin has only limited current usage in clinical medicine, but this antibiotic remains important for therapeutic and growth promotion in animals and for the bacterial disease control in plants.The increasing incidence of resistance to streptomycin and tetracyclines has been reported worldwide in Salmonella spp. of human and animal origin. Fifty-eight multidrug-resistant Salmonella enterica strains of twenty different serotypes, were chosen among the collection of multidrug-resistant strains isolated from animals and food of animal origin at the Istituto Zooprofilattico Sperimentale delle Venezie and at the Istituto Zooprofilattico Sperimentale dell’Abruzzo e Molise, during their routine surveillance activity in the 2000 and 2001 period.All strains showed resistance to at least three different antimicrobials: tetracycline and streptomycin resistances represent the 98% and 95% of the strains, respectively. Salmonella isolates were analyzed for the presence of genetic determinants conferring streptomycin and tetracyclines resistance by PCR for the tet(A and strA-strB genes, respectively.The strA-strB genes were highly prevalent in Salmonella strains of our collection, being detected in the 83% of the streptomycin resistant strains; the 68% of the tetracycline resistant strains were tet(A gene positive, indicating that this gene is widely diffused in Salmonella strains circulating in animals in Italy. Two prevalent repN- and repI1-resistance plasmids were identified in Salmonella isolates of our collection. In many strains, the strA-strB genes were linked to a particular Tn5393-derivative transposon, characterized by the presence of the insertion sequence IS1133

  20. Salmonella enterica serovar typhimurium waaP mutants show increased susceptibility to polymyxin and loss of virulence In vivo.

    Science.gov (United States)

    Yethon, J A; Gunn, J S; Ernst, R K; Miller, S I; Laroche, L; Malo, D; Whitfield, C

    2000-08-01

    In Escherichia coli, the waaP (rfaP) gene product was recently shown to be responsible for phosphorylation of the first heptose residue of the lipopolysaccharide (LPS) inner core region. WaaP was also shown to be necessary for the formation of a stable outer membrane. These earlier studies were performed with an avirulent rough strain of E. coli (to facilitate the structural chemistry required to properly define waaP function); therefore, we undertook the creation of a waaP mutant of Salmonella enterica serovar Typhimurium to assess the contribution of WaaP and LPS core phosphorylation to the biology of an intracellular pathogen. The S. enterica waaP mutant described here is the first to be both genetically and structurally characterized, and its creation refutes an earlier claim that waaP mutations in S. enterica must be leaky to maintain viability. The mutant was shown to exhibit characteristics of the deep-rough phenotype, despite its ability to produce a full-length core capped with O antigen. Further, phosphoryl modifications in the LPS core region were shown to be required for resistance to polycationic antimicrobials. The waaP mutant was significantly more sensitive to polymyxin in both wild-type and polymyxin-resistant backgrounds, despite the decreased negative charge of the mutant LPSs. In addition, the waaP mutation was shown to cause a complete loss of virulence in mouse infection models. Taken together, these data indicate that WaaP is a potential target for the development of novel therapeutic agents.

  1. Survival and recovery of Salmonella enterica serovar Typhimurium DT104 at low temperature and water activity in a broth system.

    Science.gov (United States)

    Kinsella, Kathleen J; Rowe, Tara A; Blair, Ian S; McDowell, David A; Sheridan, James J

    2006-01-01

    This study investigated the survival of Salmonella enterica serovar Typhimurium DT104 in a broth system under conditions of low temperature (4 degrees C) and low water activity (aw, 0.92 to 0.96). Incubation under these conditions resulted in significant reductions in the viability of stationary phase cells, determined by direct plating on selective XLD medium. Reductions in viable numbers were related to injury associated with initial osmotic shock (hyperosmosis) and further injury associated with longer-term storage under the above conditions. Such injured cells were, however, capable of recovering on a nonselective medium (TSA) and contributing to overall viable cell numbers in nonselective post-storage conditions. Storage at more extreme conditions, at lower aw values, led to cell death at rates influenced by storage temperature. Finally, the data obtained are considered in relation to pathogen survival on the surfaces of beef carcasses during chilling. PMID:17199519

  2. In vitro efficacy of several disinfectants against Salmonella enterica serovar Enteritidis and Escherichia coli strains from poultry

    Directory of Open Access Journals (Sweden)

    Sonia Martínez-Martínez

    2016-01-01

    Full Text Available ABSTRACT The efficacy of 28 individual or blended disinfectants against avian Salmonella enterica serovar Enteritidis and Escherichia coli strains was determined. An in vitro test in the presence and absence of serum as source of organic material was conducted. Povidone-iodine (releasing 1% available iodine, 1% potassium permanganate, 70% ethanol, 2% chlorhexidine digluconate and three commercial formulations based on quaternary ammonium compounds + formaldehyde or cresol derivates were the most effective against all strains tested and reduced bacterial counts by more than 106 times (6-log10 regardless of the presence of organic matter. These commercial compounds as well as ethanol and chlorhexidine among the individual substances tested might be helpful in the adoption of environmental control measures against these two enterobacteria in poultry industry.

  3. The Salmonella enterica PhoP directly activates the horizontally acquired SPI-2 gene sseL and is functionally different from a S. bongori ortholog.

    Directory of Open Access Journals (Sweden)

    Ohad Gal-Mor

    Full Text Available To establish a successful infection within the host, a pathogen must closely regulate multiple virulence traits to ensure their accurate temporal and spatial expression. As a highly adapted intracellular pathogen, Salmonella enterica has acquired during its evolution various virulence genes via numerous lateral transfer events, including the acquisition of the Salmonella Pathogenicity Island 2 (SPI-2 and its associated effectors. Beneficial use of horizontally acquired genes requires that their expression is effectively coordinated with the already existing virulence programs and the regulatory set-up in the bacterium. As an example for such a mechanism, we show here that the ancestral PhoPQ system of Salmonella enterica is able to regulate directly the SPI-2 effector gene sseL (encoding a secreted deubiquitinase in an SsrB-independent manner and that PhoP plays a part in a feed-forward regulatory loop, which fine-tunes the cellular level of SseL. Additionally, we demonstrate the presence of conserved cis regulatory elements in the promoter region of sseL and show direct binding of purified PhoP to this region. Interestingly, in contrast to the S. enterica PhoP, an ortholog regulator from a S. bongori SARC 12 strain was found to be impaired in promoting transcription of sseL and other genes from the PhoP regulon. These findings have led to the identification of a previously uncharacterized residue in the DNA-binding domain of PhoP, which is required for the transcriptional activation of PhoP regulated genes in Salmonella spp. Collectively our data demonstrate an interesting interface between the acquired SsrB regulon and the ancestral PhoPQ regulatory circuit, provide novel insights into the function of PhoP, and highlight a mechanism of regulatory integration of horizontally acquired genes into the virulence network of Salmonella enterica.

  4. Non-Saccharomyces yeasts protect against epithelial cell barrier disruption induced by Salmonella enterica subsp. enterica serovar Typhimurium

    DEFF Research Database (Denmark)

    Smith, Ida Mosbech; Baker, A; Arneborg, Nils;

    2015-01-01

    . 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...

  5. Targeted therapy via oral administration of attenuated Salmonella expression plasmid-vectored Stat3-shRNA cures orthotopically transplanted mouse HCC.

    Science.gov (United States)

    Tian, Y; Guo, B; Jia, H; Ji, K; Sun, Y; Li, Y; Zhao, T; Gao, L; Meng, Y; Kalvakolanu, D V; Kopecko, D J; Zhao, X; Zhang, L; Xu, D

    2012-06-01

    The development of RNA interference-based cancer gene therapies has been delayed due to the lack of effective tumor-targeting delivery systems. Attenuated Salmonella enterica serovar Typhimurium (S. Typhimurium) has a natural tropism for solid tumors. We report here the use of attenuated S. Typhimurium as a vector to deliver shRNA directly into tumor cells. Constitutively activated signal transducer and activator of transcription 3 (Stat3) is a key transcription factor involved in both hepatocellular carcinoma (HCC) growth and metastasis. In this study, attenuated S. Typhimurium was capable of delivering shRNA-expressing vectors to the targeted cancer cells and inducing RNA interference in vivo. More importantly, a single oral dose of attenuated S. Typhimurium carrying shRNA-expressing vectors targeting Stat3 induced remarkably delayed and reduced HCC (in 70% of mice). Cancer in these cured mice did not recur over 2 years following treatment. These data demonstrated that RNA interference combined with Salmonella as a delivery system may offer a novel clinical approach for cancer gene therapy. PMID:22555509

  6. Interaction of Saccharomyces boulardii with Salmonella enterica serovar Typhimurium protects mice and modifies T84 cell response to the infection.

    Directory of Open Access Journals (Sweden)

    Flaviano S Martins

    Full Text Available BACKGROUND: Salmonella pathogenesis engages host cells in two-way biochemical interactions: phagocytosis of bacteria by recruitment of cellular small GTP-binding proteins induced by the bacteria, and by triggering a pro-inflammatory response through activation of MAPKs and nuclear translocation of NF-kappaB. Worldwide interest in the use of functional foods containing probiotic bacteria for health promotion and disease prevention has increased significantly. Saccharomyces boulardii is a non-pathogenic yeast used as a probiotic in infectious diarrhea. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we reported that S. boulardii (Sb protected mice from Salmonella enterica serovar Typhimurium (ST-induced death and prevented bacterial translocation to the liver. At a molecular level, using T84 human colorectal cancer cells, we demonstrate that incubation with Sb before infection totally abolished Salmonella invasion. This correlates with a decrease of activation of Rac1. Sb preserved T84 barrier function and decreased ST-induced IL-8 synthesis. This anti-inflammatory effect was correlated with an inhibitory effect of Sb on ST-induced activation of the MAPKs ERK1/2, p38 and JNK as well as on activation of NF-kappaB. Electron and confocal microscopy experiments showed an adhesion of bacteria to yeast cells, which could represent one of the mechanisms by which Sb exerts its protective effects. CONCLUSIONS: Sb shows modulating effects on permeability, inflammation, and signal transduction pathway in T84 cells infected by ST and an in vivo protective effect against ST infection. The present results also demonstrate that Sb modifies invasive properties of Salmonella.

  7. Development of a Novel Protein Microarray Method for Serotyping Salmonella enterica Strains

    OpenAIRE

    Cai, H.Y.; Lu, L; Muckle, C A; Prescott, J F; Chen, S.

    2005-01-01

    An antibody microarray assay was developed for Salmonella serotyping based on the Kauffmann-White scheme. A model (8 by 15) array was constructed using 35 antibodies for identification of 20 common Salmonella serovars and evaluated using 117 target and 73 nontarget Salmonella strains. The assay allowed complete serovar identification of 86 target strains and partial identification of 30 target strains and allowed exclusion of the 73 nontarget strains from the target serovars.

  8. Stress Response And Pathogenesis of Salmonella enterica serovar Typhimurium

    OpenAIRE

    Shah, Jigna D.

    2011-01-01

    Salmonella is a food-borne pathogen that leads to substantial illness worldwide. The clinical syndromes associated with Salmonella infection are enteric (typhoid) fever and gastroenteritis, in healthy humans. Typhoid fever is caused by host-adapted S. Typhi and S. Paratyphi. Gastroenteritis is caused by serovars usually referred to as non typhoidal Salmonellae (NTS). In recent years, an increasing number of outbreaks due to NTS, despite increased efforts in food safety, were reported because ...

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

    Directory of Open Access Journals (Sweden)

    Cochrane Brett

    2009-12-01

    Full Text Available 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

  10. 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

  11. Lack of evidence of spill-over of Salmonella enterica between cattle and sympatric Iberian ibex (Capra pyrenaica) from a protected area in Catalonia, NE Spain.

    Science.gov (United States)

    Navarro-Gonzalez, N; Velarde, R; Porrero, M C; Mentaberre, G; Serrano, E; Mateos, A; Domínguez, L; Lavín, S

    2014-08-01

    Salmonella enterica is a zoonotic agent of worldwide importance found in a wide range of wild hosts. However, its prevalence in many popular game species has never been assessed. Iberian ibex (Capra pyrenaica) is the main game caprinae of the Iberian Peninsula and around two thousand individuals are hunted every year for trophy or for home consumption. In this work, 313 Iberian ibexes from the Ports de Tortosa i Beseit National Game Reserve (NE Spain) were tested for Salmonella enterica in faeces, and anti microbial susceptibility was determined. The exact location of shooting or capture was recorded with a GPS device to study the links of Salmonella infection with cattle presence and human proximity. Additionally, samples were taken from cattle grazing inside this reserve (n = 73). Only three Iberian ibexes (0.96%, 95% CI 0.2-2.8) were positive to Salmonella (serotype Enteritidis, Bardo and 35:r:z35), while prevalence was moderate in cattle: 21.92% (95% CI 13.10-33.14, serotype Meleagridis, Anatum, Kedougou and Othmarschen). All isolates were susceptible to the anti microbial agents tested. Moreover, a case of fatal septicaemic salmonellosis in an 11-year-old male Iberian ibex is described where Salmonella enterica serotype Enteritidis was isolated from the lung, liver and spleen samples. The low prevalence of Salmonella in Iberian ibex and the lack of shared serotypes suggest no association to cattle. Despite this, game meat aimed for human consumption should be examined, and it is strongly recommended that hunters and game keepers manipulate animals and carcasses under maximal hygienic conditions to avoid environmental contamination and human contagion.

  12. Molecular characterization of Salmonella enterica serovar 4,[5],12:i:- DT193 ASSuT strains from two outbreaks in Italy

    DEFF Research Database (Denmark)

    Barco, Lisa; Ramon, Elena; Cortini, Enzo;

    2014-01-01

    Abstract Salmonella enterica subsp. enterica serovar 4,[5],12:i:- DT193 is recognized as an emerging monophasic variant of Salmonella Typhimurium in many European countries. Resistance to ampicillin, streptomycin, sulphonamides, and tetracycline (R-type ASSuT) is described as one of the most comm...... with different restriction enzymes, MLVA, and the analysis of molecular markers related to the operon fljAB. The combination of these different molecular approaches was essential to clarify the epidemiological relationship among the strains....... profiles of resistance within this clone. Recently, strains presenting such features were isolated from two unrelated outbreaks in Italy. Strains were characterized by pulsed-field gel electrophoresis (PFGE), performed with XbaI, BlnI, and SpeI, and multiple-locus variable-number tandem repeat analysis...

  13. MANAGEMENT OF OSTEOMYELITIS CAUSED BY SALMONELLA ENTERICA SUBSP. HOUTENAE IN A TAYLOR'S CANTIL (AGKISTRODON BILINEATUS TAYLORI) USING AMIKACIN DELIVERED VIA OSMOTIC PUMP.

    Science.gov (United States)

    Clancy, Meredith M; Newton, Alisa L; Sykes, John M

    2016-06-01

    An adult female Taylor's cantil (Agkistrodon bilineatus taylori) presented with marked spinal and mandibular osteomyelitis that cultured positive for Salmonella enterica subsp. houtenae, serovar IV 43:z4,z32:-. Progression of osteomyelitis was arrested by treatment using amikacin (0.026 mg/kg per hour) delivered via subcutaneous osmotic pump for 10 mo, replacing the pump every 4 wk. No adverse effects on renal function were appreciated throughout the course of therapy. Amikacin therapy was discontinued after improvement with treatment, but 5 mo later, bony lesions worsened, and an additional abscess formed at the previous pump site. The animal's condition declined and euthanasia was elected. Postmortem examination confirmed marked osteomyelitis with Salmonella infection of same serovar as the initial biopsy. This report highlights the pathogenicity of the S. enterica subsp. houtenae serovar and the ability to deliver effective amikacin dosage via osmotic pump to arrest osteomyelitis due to salmonellosis in a venomous snake. PMID:27468053

  14. Genomic Epidemiology of Salmonella enterica Serotype Enteritidis based on Population Structure of Prevalent Lineages

    DEFF Research Database (Denmark)

    Deng, Xiangyu; Desai, Prerak T.; den Bakker, Henk C.;

    2014-01-01

    serotype Nitra strains. Single-nucleotide polymorphisms were filtered to identify 4,887 reliable loci that distinguished all isolates from each other. Our whole-genome single-nucleotide polymorphism typing approach was robust for S. enterica Enteritidis subtyping with combined data for different strains...

  15. No protective effects of high-dosage dietary zinc oxide on weaned pigs infected with Salmonella enterica serovar typhimurium DT104.

    Science.gov (United States)

    Janczyk, Pawel; Kreuzer, Susanne; Assmus, Jens; Nöckler, Karsten; Brockmann, Gudrun A

    2013-05-01

    Twenty-eight-day-old weaned pigs were fed diets with a low (LZn), medium (MZn), or high (MZn) Zn concentration (50 to 80, 150, or 2,500 mg Zn/kg of diet, respectively) provided as zinc oxide (ZnO)(24 pigs per group). They were infected orally with Salmonella enterica serovar Typhimurium DT104 on day 32. Salmonellae were cultivated from feces (up to 42 days postinfection [dpi]) and organs (2 and 42 dpi). Activation of the adaptive systemic and mucosal immune systems was investigated by recording anti-Salmonella IgG levels and levels of B and T lymphocyte subpopulations in blood and gut-associated lymphatic tissue. Growth performance was recorded as well. Salmonellae were shed at higher levels and for longer periods in the HZn group (P pig industry should be limited to 2 to 3 weeks. PMID:23435881

  16. Antibiotic exposure can induce various bacterial virulence phenotypes in multidrug-resistant Salmonella enterica serovar Typhimurium

    Science.gov (United States)

    Salmonella is one of the most prevalent bacterial foodborne diseases in the United States and causes an estimated 1 million human cases every year. Multidrug-resistant (MDR) Salmonella has emerged as a public health issue as it has been associated with increased morbidity in humans and mortality in...

  17. Development of an in vivo model for study of intestinal invasion by Salmonella enterica in chickens

    DEFF Research Database (Denmark)

    Aabo, Søren; Christensen, J.P.; Chadfield, M.S.;

    2000-01-01

    , followed by a 1-h incubation with gentamicin in order to kill noninvading bacteria. After euthanasia, Salmonella invasiveness was measured as tissue-associated counts relative to a reference strain. The ability of Salmonella invasion was 1 log(10) CFU higher per 42-mm(2) mucosal tissue in the anterior than...

  18. Characterization of quinolone resistance in Salmonella enterica serovar Indiana from chickens in China

    Science.gov (United States)

    The aim of this study was to characterize the quinolone resistance of Salmonella Indiana isolated from chickens in China. A total of 130 Salmonella isolates were obtained from chicken farms and slaughterhouses in the Shandong Province of China. All isolate serotypes were tested according to the Kauf...

  19. Evaluation of Eight Different Cephalosporins for Detection of Cephalosporin Resistance in Salmonella enterica and Escherichia coli

    NARCIS (Netherlands)

    Aarestrup, F.M.; Hasman, H.; Veldman, K.T.; Mevius, D.J.

    2010-01-01

    This study evaluates the efficacy of eight different cephalosporins for detection of cephalosporin resistance mediated by extended spectrum beta-lactamases (ESBL) and plasmidic AmpC beta-lactamases in Salmonella and Escherichia coli. A total of 138 E. coli and 86 Salmonella isolates with known beta-

  20. 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

    , which caused substantial contamination of the environment. This suggests that isolation of animals as soon as a Salmonella infection is indicated by clinical symptoms of diarrhea could be a means of reducing and controlling the spread and persistence of Salmonella in outdoor organic pig production...

  1. Monitoring of transmission of Salmonella enterica serovars in pigs using bacteriological and serological detection methods

    NARCIS (Netherlands)

    Winsen, van R.L.; Nes, van A.; Keuzenkamp, D.; Urlings, H.A.P.; Lipman, L.J.A.; Biesterveld, S.; Snijders, J.M.A.; Verheijden, J.H.M.; Knapen, van F.

    2001-01-01

    The standard method to detect Salmonella positive pigs is bacteriological examination of the faeces, but in recent years the use of Salmonella-ELISA’s have become available to screen pigs for serological evidence of infection. This study was conducted to monitor the transmission of five different Sa

  2. Quantitative comparison of intestinal invasion of zoonotic serotypes of Salmonella enterica in poultry

    DEFF Research Database (Denmark)

    Aabo, Søren; Christensen, J.P.; Chadfield, M.S.;

    2002-01-01

    The aim of the present study was to compare the invasion of selected zoonotic Salmonella serotypes of poultry in an in vivo chicken intestinal loop model and also in vitro in epithelial cell cultures. Invasion was measured relative to a reference strain, Salmonella Typhimurium 4/74 invH201::Tnpho...

  3. Selection for pro-inflammatory mediators yields chickens with increased resistance against Salmonella enterica serovar Enteritidis

    Science.gov (United States)

    Salmonella are a leading cause of foodborne illness and can be transmitted through consumption of contaminated poultry; therefore, increasing a flocks’ natural resistance to Salmonella could improve food safety. Previously, we characterized the heterophil-mediated innate immune response of two pare...

  4. Prophylactic Bacteriophage Administration More Effective than Post-infection Administration in Reducing Salmonella enterica serovar Enteritidis Shedding in Quail

    Science.gov (United States)

    Ahmadi, Mosab; Karimi Torshizi, M. Amir; Rahimi, Shaban; Dennehy, John J.

    2016-01-01

    Infections caused by Salmonella bacteria, often through poultry products, are a serious public health issue. Because of drawbacks associated with antibiotic prophylaxis, alternative treatments are sought. Bacterial viruses (bacteriophages) may provide an effective alternative, but concerns remain with respect to bacteriophage stability and effectiveness. To this end, we assessed the stability of a novel bacteriophage isolated from poultry excreta, siphovirus PSE, and its effectiveness in reducing Salmonella enterica serovar Enteritidis colonization in vitro and in vivo. Moreover, we sought to determine how the timing (prophylactic or therapeutic) and route (oral gavage or vent lip) of PSE administration impacted its effectiveness. Here we report that significant quantities of viable PSE bacteriophages were recovered following exposure to high and low pH, high temperatures, and bile salts, testifying to its ability to survive extreme conditions. In addition, we found that ileal lactic acid bacteria and Streptococcus spp. counts increased, but colibacilli and total aerobe counts decreased, in quail receiving phage PSE through both oral gavage and vent lip routes. In other experiments, we assessed the efficiency of PSE administration, in both prophylactic and therapeutic contexts, via either oral gavage or vent lip administration, on S. Enteritidis colonization of quail cecal tonsils. Our results demonstrate that administration of PSE as a preventive agent could reduce the S. Enteritidis colonization more effectively than post-challenge administration. Furthermore, oral administration of PSE phage is a more effective prophylactic tool for reduction of S. Enteritidis shedding in poultry than is vent lip administration. PMID:27555842

  5. Prophylactic Bacteriophage Administration More Effective than Post-infection Administration in Reducing Salmonella enterica serovar Enteritidis Shedding in Quail.

    Science.gov (United States)

    Ahmadi, Mosab; Karimi Torshizi, M Amir; Rahimi, Shaban; Dennehy, John J

    2016-01-01

    Infections caused by Salmonella bacteria, often through poultry products, are a serious public health issue. Because of drawbacks associated with antibiotic prophylaxis, alternative treatments are sought. Bacterial viruses (bacteriophages) may provide an effective alternative, but concerns remain with respect to bacteriophage stability and effectiveness. To this end, we assessed the stability of a novel bacteriophage isolated from poultry excreta, siphovirus PSE, and its effectiveness in reducing Salmonella enterica serovar Enteritidis colonization in vitro and in vivo. Moreover, we sought to determine how the timing (prophylactic or therapeutic) and route (oral gavage or vent lip) of PSE administration impacted its effectiveness. Here we report that significant quantities of viable PSE bacteriophages were recovered following exposure to high and low pH, high temperatures, and bile salts, testifying to its ability to survive extreme conditions. In addition, we found that ileal lactic acid bacteria and Streptococcus spp. counts increased, but colibacilli and total aerobe counts decreased, in quail receiving phage PSE through both oral gavage and vent lip routes. In other experiments, we assessed the efficiency of PSE administration, in both prophylactic and therapeutic contexts, via either oral gavage or vent lip administration, on S. Enteritidis colonization of quail cecal tonsils. Our results demonstrate that administration of PSE as a preventive agent could reduce the S. Enteritidis colonization more effectively than post-challenge administration. Furthermore, oral administration of PSE phage is a more effective prophylactic tool for reduction of S. Enteritidis shedding in poultry than is vent lip administration. PMID:27555842

  6. Genetic and Phenotypic Characterization of a Salmonella enterica serovar Enteritidis Emerging Strain with Superior Intra-macrophage Replication Phenotype

    Science.gov (United States)

    Shomer, Inna; Avisar, Alon; Desai, Prerak; Azriel, Shalhevet; Smollan, Gill; Belausov, Natasha; Keller, Nathan; Glikman, Daniel; Maor, Yasmin; Peretz, Avi; McClelland, Michael; Rahav, Galia; Gal-Mor, Ohad

    2016-01-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is one of the ubiquitous Salmonella serovars worldwide and a major cause of food-born outbreaks, which are often associated with poultry and poultry derivatives. Here we report a nation-wide S. Enteritidis clonal outbreak that occurred in Israel during the last third of 2015. Pulsed field gel electrophoresis and whole genome sequencing identified genetically related strains that were circulating in Israel as early as 2008. Global comparison linked this outbreak strain to several clinical and marine environmental isolates that were previously isolated in California and Canada, indicating that similar strains are prevalent outside of Israel. Phenotypic comparison between the 2015 outbreak strain and other clinical and reference S. Enteritidis strains showed only limited intra-serovar phenotypic variation in growth in rich medium, invasion into Caco-2 cells, uptake by J774.1A macrophages, and host cell cytotoxicity. In contrast, significant phenotypic variation was shown among different S. Enteritidis isolates when biofilm-formation, motility, invasion into HeLa cells and uptake by THP-1 human macrophages were studied. Interestingly, the 2015 outbreak clone was found to possess superior intra-macrophage replication ability within both murine and human macrophages in comparison to the other S. Enteritidis strains studied. This phenotype is likely to play a role in the virulence and host-pathogen interactions of this emerging clone.

  7. Genetically Similar Isolates of Salmonella enterica Serotype Enteritidis Persistent in China for a Long-Term Period.

    Science.gov (United States)

    Song, Qifa; Shen, Xuanyi; Yang, Yuanbin; Zhang, Danyang; Gao, Hong

    2016-07-01

    Salmonella enterica serotype Enteritidis (S. Enteritidis) is an important causative agent of nontyphoidal salmonellosis in human populations. In this study, we collected 72 S. Enteritidis strains from 2004 to 2014 in Ningbo, mid-east China. Of the 72 strains, we identified a dominant clone of 58 strains recovered from patient's feces (n = 48), blood (n = 1), pleural effusion (n = 1), chickens (n = 3), and dessert cakes (n = 5) by pulsed-field gel electrophoresis (PFGE) and variable-number of tandem repeat analysis (MLVA). The profile arrangements of MLVA were SE1-SE2-SE3-SE5-SE6-SE8-SE9: 4-4-3-11-10-1-3. These dominant strains were susceptible to ampicillin, chloramphenicol, tetracycline, ciprofloxacin, gentamicin, cefotaxime and trimethoprim-sulfamethoxazole, and resistant to nalidixic acid. Additionally, all isolates harboured virulence genes invA, sipA, sopE, and spvB when tested by PCR. Our results reveal that genetically similar S. Enteritidis strains which accounted for several outbreaks as well as blood infection and pleural cavity infection are prevalent in China for a long-term period. This situation calls for further attention in the prevention and control of foodborne disease caused by Salmonella species. PMID:27228342

  8. Sequence analysis and characterization of rolling-circle replicating plasmid pVCM01 from Salmonella enterica

    Directory of Open Access Journals (Sweden)

    Penido, A. F. B.

    2013-12-01

    Full Text Available Aims: Characterization of cryptic plasmid pVCM01 (accession number JX133088 isolated from Salmonella enterica Enteritidis. Methodology and results: The complete sequence of pVCM01 was obtained. This plasmid possesses 1981 bp, with G+C content of 57% in agreement of the range of Salmonella genomic DNA. pVCM01 has a high degree of similarity to pB and pJ plasmids. It possesses six main open reading frames, only one have a very high degree of amino acid identity with protein involved in the rolling-circle-like replication (RCR. Based on the sequence similarities, pVCM01 plasmid belonged to the pC194/pUB110 rolling-circle replicating plasmid family. The Rep pVCM01 possesses the motifs: FLTLTVRN, HPHFHTL, SGDGYVKHERW, which were present in all Rep proteins. Conclusion, significance and impact of study: The small size of pVCM01 plasmid and its stability in E. coli cells, make it an attractive candidate to develop new vectors, such as cloning and/or expression vector.

  9. Investigation of the distribution and control of Salmonella enterica serovar Enteritidis PT6 in layer breeding and egg production.

    Science.gov (United States)

    Davies, Robert; Liebana, Ernesto; Breslin, Mark

    2003-06-01

    Investigations were carried out in a layer breeder hatchery, a layer parent rearing farm, a layer parent farm and in a commercial pullet rearing and cage layer farm where Salmonella enterica serovar Enteritidis (S. Enteritidis) PT6 had become established. PT6 was initially found in focal points in the hatchery, such as hatcher ventilation ducting, tray wash areas and waste areas, but improved disinfection was followed by a rapid disappearance of contamination. Several different phage types of S. Enteritidis were found in the hatchery but most of these proved to be genotypically identical with PT6. Investigations of contaminated layer breeder and rearing sites showed that the terminal disinfection programmes in place were effective in that no carry-over of infection occurred and the organism was rapidly eliminated from the organization. Infection with PT6 originating from chicks was investigated on a commercial pullet rearing farm. After several rounds of treatment with a fluoroquinolone antibiotic and competitive exclusion, no Salmonella was found in faeces or cloacal swabs but was present in dust in one of six houses. Sampling carried out after cleaning and disinfection confirmed clearance of the organism from the site, but infection did become established in a commercial laying house receiving the birds. PMID:12850912

  10. Preparation and evaluation of immunogenic conjugates of Salmonella enterica serovar Typhi O-specific polysaccharides with diphtheria toxoid.

    Science.gov (United States)

    Ali, Aamir; An, So Jung; Cui, Changfa; Haque, Abdul; Carbis, Rodney

    2012-02-01

    Typhoid fever, caused by Salmonella enterica serovar Typhi (S. Typhi), is a major health problem particularly in developing countries. The available vaccines have certain limitations regarding their efficacy, and inability to induce an immune response especially in individuals under 2 years of age. Conjugate vaccines which consist of a bacteria-specific polysaccharide chemically bound to a carrier protein overcome these problems by inducing a T-cell dependent immune response characterized by enhanced immunogenicity in all ages. In this study, O-specific polysaccharides (OSP) of S. Typhi were conjugated to diphtheria toxoid (DT) using adipic acid dihydrazide (ADH) as a linker. These conjugates (OSP-AH-DT) were then evaluated for their immunogenicity using mice as a model and showed significantly higher levels of IgG ELISA titers (P = 0.0241 and 0.0245) than lipopolysaccharides alone. Different immunization  schedules were compared and it was found that schedule-B (three injections with 4-weeks interval) induced higher immune responses than schedule-A (three injections with 2-weeks interval). We showed that diphtheria toxoid can be successfully employed as a carrier protein for conjugation with Salmonella OSP and play an important role in facilitating adequate immune response.

  11. Application of Scutellariae radix, Gardeniae fructus, and Probiotics to Prevent Salmonella enterica Serovar Choleraesuis Infection in Swine

    Directory of Open Access Journals (Sweden)

    Chiung-Hung Chang

    2013-01-01

    Full Text Available Salmonella enterica serovar Choleraesuis, a host-adapted pathogen of swine, usually causes septicemia. Lactic acid bacteria (LAB strains have been widely studied in recent years for their probiotic properties. In this study, a mouse infection model first screened for potential agents against infection, then a pig infection model evaluated effects of LAB strains and herbal plants against infection. Scutellariae radix (SR and Gardeniae fructus (GF showed abilities to reduce bacteria shedding and suppressing serum level of TNF-α induced by infection in swine. Bioactivities of SR and GF were enhanced by combining with LAB strains, which alone could speed up the bacteria elimination time in feces and boost immunity of infected pigs. Baicalein and genipin exhibited stronger cytotoxicity than baicalin and geniposide did, as well as prevent Salmonella from invading macrophages. Our study suggests LAB strains as exhibiting multiple functions: preventing infection, enhancing immunity to prepare host defenses against further infection, and adjusting intestinal microbes’ enzymatic activity in order to convert herbal compounds to active compounds. The SR/GF-LAB strain mixture holds potential infection-prevention agents supplied as feed additives.

  12. Exposure of Salmonella enterica serovar Typhimurium to high level biocide challenge can select multidrug resistant mutants in a single step.

    Directory of Open Access Journals (Sweden)

    Rebekah N Whitehead

    Full Text Available BACKGROUND: Biocides are crucial to the prevention of infection by bacteria, particularly with the global emergence of multiply antibiotic resistant strains of many species. Concern has been raised regarding the potential for biocide exposure to select for antibiotic resistance due to common mechanisms of resistance, notably efflux. METHODOLOGY/PRINCIPAL FINDINGS: Salmonella enterica serovar Typhimurium was challenged with 4 biocides of differing modes of action at both low and recommended-use concentration. Flow cytometry was used to investigate the physiological state of the cells after biocide challenge. After 5 hours exposure to biocide, live cells were sorted by FACS and recovered. Cells recovered after an exposure to low concentrations of biocide had antibiotic resistance profiles similar to wild-type cells. Live cells were recovered after exposure to two of the biocides at in-use concentration for 5 hours. These cells were multi-drug resistant and accumulation assays demonstrated an efflux phenotype of these mutants. Gene expression analysis showed that the AcrEF multidrug efflux pump was de-repressed in mutants isolated from high-levels of biocide. CONCLUSIONS/SIGNIFICANCE: These data show that a single exposure to the working concentration of certain biocides can select for mutant Salmonella with efflux mediated multidrug resistance and that flow cytometry is a sensitive tool for identifying biocide tolerant mutants. The propensity for biocides to select for MDR mutants varies and this should be a consideration when designing new biocidal formulations.

  13. No Protective Effects of High-Dosage Dietary Zinc Oxide on Weaned Pigs Infected with Salmonella enterica Serovar Typhimurium DT104

    OpenAIRE

    Janczyk, Pawel; Kreuzer, Susanne; Assmus, Jens; Nöckler, Karsten; Gudrun A Brockmann

    2013-01-01

    Twenty-eight-day-old weaned pigs were fed diets with a low (LZn), medium (MZn), or high (MZn) Zn concentration (50 to 80, 150, or 2,500 mg Zn/kg of diet, respectively) provided as zinc oxide (ZnO)(24 pigs per group). They were infected orally with Salmonella enterica serovar Typhimurium DT104 on day 32. Salmonellae were cultivated from feces (up to 42 days postinfection [dpi]) and organs (2 and 42 dpi). Activation of the adaptive systemic and mucosal immune systems was investigated by recordi...

  14. Prevalence and Genetic Properties of Salmonella enterica Serovar Typhimurium Definitive Phage Type 104 Isolated from Rattus norvegicus and Rattus rattus House Rats in Yokohama City, Japan▿ †

    OpenAIRE

    Yokoyama, Eiji; Maruyama, Soichi; Kabeya, Hidenori; Hara, Siro; Sata, Shin; KUROKI, Toshiro; Yamamoto, Tomoko

    2007-01-01

    Salmonella enterica serovar Typhimurium was isolated from the intestinal contents of Rattus rattus and Rattus norvegicus house rats captured at two buildings, designated buildings J and YS, in Yokohama City, Japan. From October 1997 to September 1998, 52 of 339 (15.3%) house rats were found to carry Salmonella serovar Typhimurium definitive phage type 104 (DT104). In building J, 26 of 161 (16.1%) house rats carried DT104 over the 1-year study period, compared to 26 of 178 (14.6%) rats in buil...

  15. Broilers with low serum Mannose-binding Lectin show increased faecal shedding of Salmonella enterica serovar Montevideo

    DEFF Research Database (Denmark)

    Ulrich-Lynge, Sofie Louise; Juul-Madsen, Helle Risdahl; Kjærup, Rikke Brødsgaard;

    2016-01-01

    concentration indicate that chicken MBL affects Salmonella resistance; further studies are imperative in conventional broiler chickens. In this study 104 conventional day-old chickens (offspring from a cross between Cobb 500 male and female parent breeders) were orally infected with Salmonella enterica subsp...... the semi-quantitative real-time PCR method from L/L chickens than from the L/H chickens at week 5 pi. These results indicate that genetically determined basic levels of MBL may influence S. Montevideo susceptibility....

  16. Co-occurrence of ACSSuT and cephalosporin resistance phenotypes is mediated by int1-associated elements in nontyphoidal Salmonella enterica from human infections in Spain.

    Science.gov (United States)

    Campos, Maria Jorge; Palomo, Gonzalo; Hormeño, Lorena; Ugarte, María; Porrero, María Concepción; Herrera-León, Silvia; Vadillo, Santiago; Píriz, Segundo; Quesada, Alberto

    2013-10-01

    A screening of antimicrobial resistance and its genetic determinants has been performed on 300 Salmonella enterica isolates collected during 2004-2008 from human infections in Spain. Salmonella Typhimurium and Salmonella Enteritidis were the major serotypes, which were found with similar frequencies covering 80% of the bacterial collection. Salmonella Typhimurium isolates frequently shared low susceptibility to antimicrobials of the penta-resistance phenotype (ACSSuT) and/or cephalosporin resistance. The ACSSuT profile was found closely linked to int1-associated gene cassettes, with major elements carrying DNA fragments of 1.0 Kb (aadA2 gene) plus 1.2 Kb (blaPSE-1 gene) or 2.0 Kb (aadA1 and blaOXA-1 genes). Among these, ACSSuT and cephalosporin resistances were associated in Salmonella Typhimurium isolates expressing the blaOXA gene. β-lactamase activities were also detected from isolates carrying blaTEM, blaCMY, or blaSHV, although only the two last genes expressed extended-spectrum β-lactamases. The clonal analysis of S. enterica strains suggests that both horizontal and vertical transfer mechanisms are involved in the wide dissemination of their antimicrobial resistance.

  17. Multi-locus variable-number tandem repeat profiling of Salmonella enterica serovar Typhi isolates from blood cultures and gallbladder specimens from Makassar, South-Sulawesi, Indonesia.

    Directory of Open Access Journals (Sweden)

    Mochammad Hatta

    Full Text Available Multi-locus variable-number tandem repeat analysis differentiated 297 Salmonella enterica serovar Typhi blood culture isolates from Makassar in 76 genotypes and a single unique S. Typhi genotype was isolated from the cholecystectomy specimens of four patients with cholelithiasis. The high diversity in S. Typhi genotypes circulating in Makassar indicates that the number of carriers could be very large, which may complicate disease prevention and control.

  18. Septic arthritis of the hip in a Cambodian child caused by multidrug-resistant Salmonella enterica serovar Typhi with intermediate susceptibility to ciprofloxacin treated with ceftriaxone and azithromycin.

    Science.gov (United States)

    Pocock, J M; Khun, P A; Moore, C E; Vuthy, S; Stoesser, N; Parry, C M

    2014-08-01

    Septic arthritis is a rare complication of typhoid fever. A 12-year-old boy without pre-existing disease attended a paediatric hospital in Cambodia with fever and left hip pain. A hip synovial fluid aspirate grew multidrug-resistant Salmonella enterica ser. Typhi with intermediate susceptibility to ciprofloxacin. Arthrotomy, 2 weeks of intravenous ceftriaxone and 4 weeks of oral azithromycin led to resolution of symptoms. The optimum management of septic arthritis in drug-resistant typhoid is undefined.

  19. Proteolytic Inhibition of Salmonella enterica Serovar Typhimurium-Induced Activation of the Mitogen-Activated Protein Kinases ERK and JNK in Cultured Human Intestinal Cells

    OpenAIRE

    Mynott, Tracey L.; Crossett, Ben; Prathalingam, S. Radhika

    2002-01-01

    Bromelain, a mixture of cysteine proteases from pineapple stems, blocks signaling by the mitogen-activated protein (MAP) kinases extracellular regulated kinase 1 (ERK-1) and ERK-2, inhibits inflammation, and protects against enterotoxigenic Escherichia coli infection. In this study, we examined the effect of bromelain on Salmonella enterica serovar Typhimurium infection, since an important feature of its pathogenesis is its ability to induce activation of ERK-1 and ERK-2, which leads to inter...

  20. Prevalence of shiga toxin producing Escherichia coli, Salmonella enterica, and Listeria monocytogenes at public access watershed sites in a California Central Coast agricultural region

    OpenAIRE

    Cooley, Michael B.; Quiñones, Beatriz; Oryang, David; Mandrell, Robert E.; Gorski, Lisa

    2014-01-01

    Produce contaminated with enteric pathogens is a major source of foodborne illness in the United States. Lakes, streams, rivers, and ponds were sampled with Moore swabs bi-monthly for over 2 years at 30 locations in the vicinity of a leafy green growing region on the Central California Coast and screened for Shiga toxin producing Escherichia coli (STEC), Salmonella enterica, and Listeria monocytogenes to evaluate the prevalence and persistence of pathogen subtypes. The prevalence of STEC from...

  1. Prevalence of enterobacteriaceae in Tupinambis merianae (Squamata: Teiidae) from a captive facility in Central Brazil, with a profile of antimicrobial drug resistance in Salmonella enterica

    OpenAIRE

    Andréa de Moraes Carvalho; Ayrton Klier Péres Júnior; Maria Auxiliadora Andrade; Valéria de Sá Jayme

    2013-01-01

    The present study reports the presence of enterobacteriaceae in Tegu Lizards (Tupinambis merianae)from a captive facility in central Brazil. From a total of 30 animals, 10 juveniles and 20 adults (10 males, 10 females), 60 samples were collected, in two periods separated by 15 days. The samples were cultivated in Xylose-lysine-deoxycholate agar (XLT4) and MacConkey agar. The Salmonella enterica were tested for antimicrobial susceptibility. A total of 78 bacteria was isolated, of wich 27 were ...

  2. Influence of a Probiotic Strain of Enterococcus faecium on Salmonella enterica Serovar Typhimurium DT104 Infection in a Porcine Animal Infection Model▿

    OpenAIRE

    Szabó, István; Lothar H Wieler; Tedin, Karsten; Scharek-Tedin, Lydia; Taras, David; Hensel, Andreas; Appel, Bernd; Nöckler, Karsten

    2009-01-01

    The beneficial effects of probiotic Enterococcus spp. in different hosts, such as mice and humans, have previously been reported in several studies. However, studies of large domestic animals, as well as challenge studies with pathogenic microorganisms, are very rare. Here, we investigated the influence of oral treatment of pigs with the probiotic bacterium Enterococcus faecium NCIMB 10415 on Salmonella enterica serovar Typhimurium DT104 infections in weaning piglets. Clinical symptoms, fecal...

  3. Salmonella enterica Serovar Typhi Modulates Cell Surface Expression of Its Receptor, the Cystic Fibrosis Transmembrane Conductance Regulator, on the Intestinal Epithelium

    OpenAIRE

    Lyczak, Jeffrey B.; Pier, Gerald B.

    2002-01-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) protein is an epithelial receptor mediating the translocation of Salmonella enterica serovar Typhi to the gastric submucosa. Since the level of cell surface CFTR is directly related to the efficiency of serovar Typhi translocation, the goal of this study was to measure CFTR expression by the intestinal epithelium during infection. CFTR protein initially present in the epithelial cell cytoplasm was rapidly trafficked to the plasma ...

  4. Multiplex T-RFLP Allows for Increased Target Number and Specificity: Detection of Salmonella enterica and Six Species of Listeria in a Single Test

    OpenAIRE

    Elliott, Geoffrey N.; Nadine Thomas; Marion Macrae; Campbell, Colin D.; Ogden, Iain D; Singh, Brajesh K.

    2012-01-01

    A multiplex T-RFLP test was developed to detect and identify Salmonella enterica and all six species of Listeria inoculated into milk at minimal levels. Extensive in silico analysis was used to design a fifteen-primer, six-amplimer methodology and in vitro application showed target organism DNA, when amplified individually, yielded the predicted terminal restriction fragments (TRFs) following digestion. Non-target organisms were either not-amplified or yielded TRFs which did not interfere wit...

  5. Association of virulence plasmid and antibiotic resistance determinants with chromosomal multilocus genotypes in Mexican Salmonella enterica serovar Typhimurium strains

    Directory of Open Access Journals (Sweden)

    Silva Claudia

    2009-07-01

    Full Text Available Abstract Background Bacterial genomes are mosaic structures composed of genes present in every strain of the same species (core genome, and genes present in some but not all strains of a species (accessory genome. The aim of this study was to compare the genetic diversity of core and accessory genes of a Salmonella enterica subspecies enterica serovar Typhimurium (Typhimurium population isolated from food-animal and human sources in four regions of Mexico. Multilocus sequence typing (MLST and macrorestriction fingerprints by pulsed-field gel electrophoresis (PFGE were used to address the core genetic variation, and genes involved in pathogenesis and antibiotic resistance were selected to evaluate the accessory genome. Results We found a low genetic diversity for both housekeeping and accessory genes. Sequence type 19 (ST19 was supported as the founder genotype of STs 213, 302 and 429. We found a temporal pattern in which the derived ST213 is replacing the founder ST19 in the four geographic regions analyzed and a geographic trend in the number of resistance determinants. The distribution of the accessory genes was not random among chromosomal genotypes. We detected strong associations among the different accessory genes and the multilocus chromosomal genotypes (STs. First, the Salmonella virulence plasmid (pSTV was found mostly in ST19 isolates. Second, the plasmid-borne betalactamase cmy-2 was found only in ST213 isolates. Third, the most abundant integron, IP-1 (dfrA12, orfF and aadA2, was found only in ST213 isolates. Fourth, the Salmonella genomic island (SGI1 was found mainly in a subgroup of ST19 isolates carrying pSTV. The mapping of accessory genes and multilocus genotypes on the dendrogram derived from macrorestiction fingerprints allowed the establishment of genetic subgroups within the population. Conclusion Despite the low levels of genetic diversity of core and accessory genes, the non-random distribution of the accessory genes

  6. [Investigation of pathogenic phenotypes and virulence determinants of food-borne Salmonella enterica strains in Caenorhabditis elegans animal model].

    Science.gov (United States)

    Aksoy, Deniz; Şen, Ece

    2015-10-01

    Salmonellosis, caused by non-typhoidal Salmonella enterica serovars with the consumption of contaminated food, is one of the leading food-borne disease that makes microbial food safety an important public health issue. This study was performed in order to determine the antibiotic resistance, serotyping, plasmid profiles and pathogenicity potentials of food-borne Salmonella isolates in Caenorhabditis elegans animal model system in Edirne province, located at Thrace region of Turkey. In this study, 32 Salmonella isolates, of which 26 belonged to Infantis, four to Enteritidis, one to Telaviv and one to Kentucky serovars, isolated from chicken carcasses were used. Antibiotic resistance profiles were determined by disc diffusion and broth microdilution methods. A new C.elegans nematode animal model system was used to determine the pathogenicity potential of the isolates. The antibiotic resistance profiles revealed that one (3.1%) isolate was resistant to gentamicin, two (6.2%) to ciprofloxacin, three (9.4%) to ampicillin, 18 (56.3%) to kanamycin, 19 (60.8%) to neomycin, 25 (78.1%) to tetracycline, 25 (78.1%) to trimethoprim, 26 (81.25%) to nalidixic acid, 27 (84.4%) to streptomycin and 32 (100%) to sulfonamide. All of the 32 strains were susceptible to chloramphenicol and ampicillin/sulbactam. High levels of resistance to streptomycin, nalidixic acid, tetracycline, trimethoprim, sulfonamide, kanamycin and neomycin was determined. According to the plasmid analysis, six isolates (18.75%) harboured 1-3 plasmids with sizes between 1.2 and 42.4 kb. In C.elegans nematode animal model system, the time (in days) required to kill 50% (TD50) of nematodes was calculated for each experimental group. TD50 values of the nematode group fed with S.Typhimurium ATCC 14028 that was used as the positive control and another group fed with E.coli OP50 as the negative control were 4.2 ± 0.5 days and 8.0 ± 0.02 days, respectively. TD50 of the groups fed with Salmonella isolates ranged

  7. Anti-angiogenesis Effect on Glioma of Attenuated Salmonella Typhimurium Vaccine Strain with flk-1 Gene

    Institute of Scientific and Technical Information of China (English)

    冯珂珂; 赵洪洋; 陈剑; 姚东晓; 姜小兵; 周伟

    2004-01-01

    To investigate the anti-vasculature effects and the anti-glioma effects of attenuated Salmonella typhimurium vaccine strain expressing VEGFR2 (flk-1) gene, plasmid pcDNA3. 1-flk1 was constructed and electro-transfected into live attenuated Salmonella typhimurium strain SL7207. Mouse models of intracranial Gl261 glioblastoma were treated with an orally administered attenuated Salmonella typhimurium expressing flk-1 gene. The survival period was recorded and vessel density was observed by immunofluorescence. CTLs activity was measured by MTT assay.Our results showed that attenuated Salmonella typhimurium vaccine strain expressing flk-1 gene could significantly inhibit glioblastoma growth, reduce vessel density, prolong the survival period and improve the survival rate in these mice. The flk-1 specific CTLs activity was increased obviously after the vaccination. Our study showed that attenuated Salmonella typhimurium vaccine strain expressing flk-1 gene could break peripheral immune tolerance a in glioma gainst this self-antigen and kill endothelial cells by the orally administered vaccine and can be used for both prophylactic and therapeutic purposes.

  8. A Genomic Island in Salmonella enterica ssp. salamae provides new insights on the genealogy of the locus of enterocyte effacement.

    Directory of Open Access Journals (Sweden)

    P Scott Chandry

    Full Text Available The genomic island encoding the locus of enterocyte effacement (LEE is an important virulence factor of the human pathogenic Escherichia coli. LEE typically encodes a type III secretion system (T3SS and secreted effectors capable of forming attaching and effacing lesions. Although prominent in the pathogenic E. coli such as serotype O157:H7, LEE has also been detected in Citrobacter rodentium, E. albertii, and although not confirmed, it is likely to also be in Shigella boydii. Previous phylogenetic analysis of LEE indicated the genomic island was evolving through stepwise acquisition of various components. This study describes a new LEE region from two strains of Salmonella enterica subspecies salamae serovar Sofia along with a phylogenetic analysis of LEE that provides new insights into the likely evolution of this genomic island. The Salmonella LEE contains 36 of the 41 genes typically observed in LEE within a genomic island of 49, 371 bp that encodes a total of 54 genes. A phylogenetic analysis was performed on the entire T3SS and four T3SS genes (escF, escJ, escN, and escV to elucidate the genealogy of LEE. Phylogenetic analysis inferred that the previously known LEE islands are members of a single lineage distinct from the new Salmonella LEE lineage. The previously known lineage of LEE diverged between islands found in Citrobacter and those in Escherichia and Shigella. Although recombination and horizontal gene transfer are important factors in the genealogy of most genomic islands, the phylogeny of the T3SS of LEE can be interpreted with a bifurcating tree. It seems likely that the LEE island entered the Enterobacteriaceae through horizontal gene transfer as a single unit, rather than as separate subsections, which was then subjected to the forces of both mutational change and recombination.

  9. A Genomic Island in Salmonella enterica ssp. salamae provides new insights on the genealogy of the locus of enterocyte effacement.

    Science.gov (United States)

    Chandry, P Scott; Gladman, Simon; Moore, Sean C; Seemann, Torsten; Crandall, Keith A; Fegan, Narelle

    2012-01-01

    The genomic island encoding the locus of enterocyte effacement (LEE) is an important virulence factor of the human pathogenic Escherichia coli. LEE typically encodes a type III secretion system (T3SS) and secreted effectors capable of forming attaching and effacing lesions. Although prominent in the pathogenic E. coli such as serotype O157:H7, LEE has also been detected in Citrobacter rodentium, E. albertii, and although not confirmed, it is likely to also be in Shigella boydii. Previous phylogenetic analysis of LEE indicated the genomic island was evolving through stepwise acquisition of various components. This study describes a new LEE region from two strains of Salmonella enterica subspecies salamae serovar Sofia along with a phylogenetic analysis of LEE that provides new insights into the likely evolution of this genomic island. The Salmonella LEE contains 36 of the 41 genes typically observed in LEE within a genomic island of 49, 371 bp that encodes a total of 54 genes. A phylogenetic analysis was performed on the entire T3SS and four T3SS genes (escF, escJ, escN, and escV) to elucidate the genealogy of LEE. Phylogenetic analysis inferred that the previously known LEE islands are members of a single lineage distinct from the new Salmonella LEE lineage. The previously known lineage of LEE diverged between islands found in Citrobacter and those in Escherichia and Shigella. Although recombination and horizontal gene transfer are important factors in the genealogy of most genomic islands, the phylogeny of the T3SS of LEE can be interpreted with a bifurcating tree. It seems likely that the LEE island entered the Enterobacteriaceae through horizontal gene transfer as a single unit, rather than as separate subsections, which was then subjected to the forces of both mutational change and recombination.

  10. The influence of autophagy on mouse inflammatory responses caused by Salmonella enterica serovar Typhimurium with spv genes

    Institute of Scientific and Technical Information of China (English)

    LI Yuan-Yuan; WU Shu-Yan; CHU Yuan-Yuan; LIAO LI; LIQiong; HUANG Rui

    2011-01-01

    An investigation into the effects of Salmonella plasmid virulence genes (spv) on autophagy,apoptosis,and inflammation was carried out in mice,using a strain of Salmonella enterica serovar Typhimurium (S.typhimurium) SR-11 carrying spv.Strain BRD509 without spy was used as a control.Results showed that the expression of autophagy protein Beclin-1 in the livers and spleens in the SR-11 group was lower than that in the BRD509 group,while the apoptosis protein,Caspase-3,was higher in the SR-11 group.Inflammatory cytokine levels [interleukin 12 (IL-12) and interferon γ (IFN-γ)] were higher in the SR-11 group compared with those in the BRD509 group since 4 d post-infection.In addition,we found an increase in severe pathological changes and larger viable bacterial amounts in livers and spleens in the SR-11 group.After intervention with autophagy agonist rapamycin (RAPA),Beclin-1 expression increased in both groups,while Caspase-3 expression was different between the two groups: Caspase-3 decreased in the SR-11 group but increased in the BRD509 group.Moreover,RAPA decreased cytokine levels,bacterial quantity and organ-related injury in the SR-11 group whereas RAPA increased cytokine levels and aggravated organ injury in the BRD509 group.Results from these studies suggest that S.typhimurium with spv genes may exacerbate infection by inhibiting autophagy and affecting the production of inflammatory cytokines.RAPA-enhanced autophagy may improve the secretion of cytokines in order to protect the host from damaging by Salmonella infection.Our study suggests that the regulation of cellular autophagy may play a role in the prevention and control of certain infectious diseases.

  11. Fifteen years of successful spread of Salmonella enterica serovar Mbandaka clone ST413 in Poland and its public health consequences

    Directory of Open Access Journals (Sweden)

    Andrzej Hoszowski

    2016-06-01

    Full Text Available In the 1990s, [i]Salmonella enterica[/i] serovar (S. Mbandaka occurred in feed and poultry in Poland. In the following years, the serovar also gained epidemiological importance in other EU countries. The objectives of current study were to evaluate the genetic relationship of contemporary S. Mbandaka with isolates originating from the beginning of the epidemics, and to assess the contribution of poultry as the source of infections in humans. Seventy S. Mbandaka isolated mainly in 2009 – 2010 from humans, poultry, food, and feed were typed with API ID32 [sup]®[/sup], MIC, plasmid profiling, PFGE, and MLST. PCR and sequencing were used to identify plasmid mediated quinolone and cephalosporin resistance mechanisms. Six biochemical profiles were identified and 59 of S. Mbandaka proved to be susceptible to the applied antimicrobials. Eight strains carried plasmids and a few of them were positive for [i]bla[/i][sub]CMY-2[/sub] and [i]qnr[/i]S1 genes. Two clusters of 15 [i]XbaI[/i]-PFGE profiles with similarity of 77.5% were found. The first cluster, gathered 7 profiles involving historical isolates and several contemporary non-human S. Mbandaka. The predominant profile in the second cluster consisted of 28 human and 1 broiler isolate. MLST analysis showed sequence type ST413 occurring among all tested isolates. The identification of close genetic relationships between S. Mbandaka of human and poultry origin indicates animals as a primal human infection route. Despite [i]Salmonella [/i]control programmes, the S. Mbandaka ST413 clone has been circulating for several years in Poland. [i]Salmonella[/i] control polices in food production chain should be continuously updated to target serovars of major epidemiological importance. Resistance noted in S. Mbandaka to such antimicrobials as fluoroquinolones and cephalosporins may hinder public health.

  12. Fifteen years of successful spread of Salmonella enterica serovar Mbandaka clone ST413 in Poland and its public health consequences.

    Science.gov (United States)

    Hoszowski, Andrzej; Zając, Magdalena; Lalak, Anna; Przemyk, Paweł; Wasyl, Dariusz

    2016-06-01

    In the 1990s, Salmonella enterica serovar (S.) Mbandaka occurred in feed and poultry in Poland. In the following years, the serovar also gained epidemiological importance in other EU countries. The objectives of current study were to evaluate the genetic relationship of contemporary S. Mbandaka with isolates originating from the beginning of the epidemics, and to assess the contribution of poultry as the source of infections in humans. Seventy S. Mbandaka isolated mainly in 2009 - 2010 from humans, poultry, food, and feed were typed with API ID32 (®), MIC, plasmid profiling, PFGE, and MLST. PCR and sequencing were used to identify plasmid mediated quinolone and cephalosporin resistance mechanisms. Six biochemical profiles were identified and 59 of S. Mbandaka proved to be susceptible to the applied antimicrobials. Eight strains carried plasmids and a few of them were positive for blaCMY-2 and qnrS1 genes. Two clusters of 15 XbaI-PFGE profiles with similarity of 77.5% were found. The first cluster, gathered 7 profiles involving historical isolates and several contemporary non-human S. Mbandaka. The predominant profile in the second cluster consisted of 28 human and 1 broiler isolate. MLST analysis showed sequence type ST413 occurring among all tested isolates. The identification of close genetic relationships between S. Mbandaka of human and poultry origin indicates animals as a primal human infection route. Despite Salmonella control programmes, the S. Mbandaka ST413 clone has been circulating for several years in Poland. Salmonella control polices in food production chain should be continuously updated to target serovars of major epidemiological importance. Resistance noted in S. Mbandaka to such antimicrobials as fluoroquinolones and cephalosporins may hinder public health. PMID:27294625

  13. Regulation of the AcrAB multidrug efflux pump in Salmonella enterica serovar Typhimurium in response to indole and paraquat.

    Science.gov (United States)

    Nikaido, Eiji; Shirosaka, Ikue; Yamaguchi, Akihito; Nishino, Kunihiko

    2011-03-01

    Salmonella enterica serovar Typhimurium has at least nine multidrug efflux pumps. Among these, AcrAB is constitutively expressed and is the most efficient, playing a role in both drug resistance and virulence. The acrAB locus is induced by indole, Escherichia coli-conditioned medium, and bile salts. This induction is dependent on RamA through the binding sequence in the upstream region of acrA that binds RamA. In the present study, we made a detailed investigation of the ramA and acrAB induction mechanisms in Salmonella in response to indole, a biological oxidant for bacteria. We found that acrAB and ramA induction in response to indole is dependent on RamR. However, the cysteine residues of RamR do not play a role in the induction of ramA in response to indole, and the oxidative effect of indole is therefore not related to ramA induction via RamR. Furthermore, we showed that paraquat, a superoxide generator, induces acrAB but not ramA. We further discovered that the mechanism of acrAB induction in response to paraquat is dependent on SoxS. The data indicate that there are at least two independent induction pathways for acrAB in response to extracellular signals such as indole and paraquat. We propose that Salmonella utilizes these regulators for acrAB induction in response to extracellular signals in order to adapt itself to environmental conditions. PMID:21148208

  14. Whole Genome DNA Sequence Analysis of Salmonella subspecies enterica serotype Tennessee obtained from related peanut butter foodborne outbreaks.

    Science.gov (United States)

    Wilson, Mark R; Brown, Eric; Keys, Chris; Strain, Errol; Luo, Yan; Muruvanda, Tim; Grim, Christopher; Jean-Gilles Beaubrun, Junia; Jarvis, Karen; Ewing, Laura; Gopinath, Gopal; Hanes, Darcy; Allard, Marc W; Musser, Steven

    2016-01-01

    Establishing an association between possible food sources and clinical isolates requires discriminating the suspected pathogen from an environmental background, and distinguishing it from other closely-related foodborne pathogens. We used whole genome sequencing (WGS) to Salmonella subspecies enterica serotype Tennessee (S. Tennessee) to describe genomic diversity across the serovar as well as among and within outbreak clades of strains associated with contaminated peanut butter. We analyzed 71 isolates of S. Tennessee from disparate food, environmental, and clinical sources and 2 other closely-related Salmonella serovars as outgroups (S. Kentucky and S. Cubana), which were also shot-gun sequenced. A whole genome single nucleotide polymorphism (SNP) analysis was performed using a maximum likelihood approach to infer phylogenetic relationships. Several monophyletic lineages of S. Tennessee with limited SNP variability were identified that recapitulated several food contamination events. S. Tennessee clades were separated from outgroup salmonellae by more than sixteen thousand SNPs. Intra-serovar diversity of S. Tennessee was small compared to the chosen outgroups (1,153 SNPs), suggesting recent divergence of some S. Tennessee clades. Analysis of all 1,153 SNPs structuring an S. Tennessee peanut butter outbreak cluster revealed that isolates from several food, plant, and clinical isolates were very closely related, as they had only a few SNP differences between them. SNP-based cluster analyses linked specific food sources to several clinical S. Tennessee strains isolated in separate contamination events. Environmental and clinical isolates had very similar whole genome sequences; no markers were found that could be used to discriminate between these sources. Finally, we identified SNPs within variable S. Tennessee genes that may be useful markers for the development of rapid surveillance and typing methods, potentially aiding in traceback efforts during future

  15. Whole Genome DNA Sequence Analysis of Salmonella subspecies enterica serotype Tennessee obtained from related peanut butter foodborne outbreaks.

    Directory of Open Access Journals (Sweden)

    Mark R Wilson

    Full Text Available Establishing an association between possible food sources and clinical isolates requires discriminating the suspected pathogen from an environmental background, and distinguishing it from other closely-related foodborne pathogens. We used whole genome sequencing (WGS to Salmonella subspecies enterica serotype Tennessee (S. Tennessee to describe genomic diversity across the serovar as well as among and within outbreak clades of strains associated with contaminated peanut butter. We analyzed 71 isolates of S. Tennessee from disparate food, environmental, and clinical sources and 2 other closely-related Salmonella serovars as outgroups (S. Kentucky and S. Cubana, which were also shot-gun sequenced. A whole genome single nucleotide polymorphism (SNP analysis was performed using a maximum likelihood approach to infer phylogenetic relationships. Several monophyletic lineages of S. Tennessee with limited SNP variability were identified that recapitulated several food contamination events. S. Tennessee clades were separated from outgroup salmonellae by more than sixteen thousand SNPs. Intra-serovar diversity of S. Tennessee was small compared to the chosen outgroups (1,153 SNPs, suggesting recent divergence of some S. Tennessee clades. Analysis of all 1,153 SNPs structuring an S. Tennessee peanut butter outbreak cluster revealed that isolates from several food, plant, and clinical isolates were very closely related, as they had only a few SNP differences between them. SNP-based cluster analyses linked specific food sources to several clinical S. Tennessee strains isolated in separate contamination events. Environmental and clinical isolates had very similar whole genome sequences; no markers were found that could be used to discriminate between these sources. Finally, we identified SNPs within variable S. Tennessee genes that may be useful markers for the development of rapid surveillance and typing methods, potentially aiding in traceback efforts

  16. Whole Genome DNA Sequence Analysis of Salmonella subspecies enterica serotype Tennessee obtained from related peanut butter foodborne outbreaks.

    Science.gov (United States)

    Wilson, Mark R; Brown, Eric; Keys, Chris; Strain, Errol; Luo, Yan; Muruvanda, Tim; Grim, Christopher; Jean-Gilles Beaubrun, Junia; Jarvis, Karen; Ewing, Laura; Gopinath, Gopal; Hanes, Darcy; Allard, Marc W; Musser, Steven

    2016-01-01

    Establishing an association between possible food sources and clinical isolates requires discriminating the suspected pathogen from an environmental background, and distinguishing it from other closely-related foodborne pathogens. We used whole genome sequencing (WGS) to Salmonella subspecies enterica serotype Tennessee (S. Tennessee) to describe genomic diversity across the serovar as well as among and within outbreak clades of strains associated with contaminated peanut butter. We analyzed 71 isolates of S. Tennessee from disparate food, environmental, and clinical sources and 2 other closely-related Salmonella serovars as outgroups (S. Kentucky and S. Cubana), which were also shot-gun sequenced. A whole genome single nucleotide polymorphism (SNP) analysis was performed using a maximum likelihood approach to infer phylogenetic relationships. Several monophyletic lineages of S. Tennessee with limited SNP variability were identified that recapitulated several food contamination events. S. Tennessee clades were separated from outgroup salmonellae by more than sixteen thousand SNPs. Intra-serovar diversity of S. Tennessee was small compared to the chosen outgroups (1,153 SNPs), suggesting recent divergence of some S. Tennessee clades. Analysis of all 1,153 SNPs structuring an S. Tennessee peanut butter outbreak cluster revealed that isolates from several food, plant, and clinical isolates were very closely related, as they had only a few SNP differences between them. SNP-based cluster analyses linked specific food sources to several clinical S. Tennessee strains isolated in separate contamination events. Environmental and clinical isolates had very similar whole genome sequences; no markers were found that could be used to discriminate between these sources. Finally, we identified SNPs within variable S. Tennessee genes that may be useful markers for the development of rapid surveillance and typing methods, potentially aiding in traceback efforts during future

  17. Oral Vaccination with Salmonella enterica as a Cruzipain-DNA Delivery System Confers Protective Immunity against Trypanosoma cruzi▿

    Science.gov (United States)

    Cazorla, Silvia I.; Becker, Pablo D.; Frank, Fernanda M.; Ebensen, Thomas; Sartori, María J.; Corral, Ricardo S.; Malchiodi, Emilio L.; Guzmán, Carlos A.

    2008-01-01

    To stimulate both local and systemic immune responses against Trypanosoma cruzi, Salmonella enterica serovar Typhimurium aroA was exploited as a DNA delivery system for cruzipain (SCz). In a murine model we compared SCz alone (GI) or coadministered with Salmonella carrying a plasmid encoding granulocyte-macrophage colony-stimulating factor (GII), as well as protocols in which SCz priming was followed by boosting with recombinant cruzipain (rCz) admixed with either CpG-ODN (GIII) or MALP-2, a synthetic derivative of a macrophage-activating lipopeptide of 2 kDa from Mycoplasma fermentans (GIV). The results showed that protocols that included four oral doses of SCz (GI) elicited mainly a mucosal response characterized by immunoglobulin A (IgA) secretion and proliferation of gut-associated lymphoid tissue cells, with weak systemic responses. In contrast, the protocol that included a boost with rCz plus CpG (GIII) triggered stronger systemic responses in terms of Cz-specific serum IgG titers, splenocyte proliferation, gamma interferon (IFN-γ) secretion, and delayed-type hypersensitivity response. Trypomastigote challenge of vaccinated mice resulted in significantly lower levels of parasitemia compared to controls. Protection was abolished by depletion of either CD4+ or CD8+ T cells. Parasite control was also evident from the reduction of tissue damage, as revealed by histopathologic studies and serum levels of enzymes that are markers of muscle injury in chronic Chagas' disease (i.e., creatine kinase, aspartate aminotransferase, and lactate dehydrogenase). Enhanced release of IFN-γ and interleukin-2 was observed in GI and GII upon restimulation of splenocytes in the nonparasitic phase of infection. Our results indicate that Salmonella-mediated delivery of Cz-DNA by itself promotes the elicitation of an immune response that controls T. cruzi infection, thereby reducing parasite loads and subsequent damage to muscle tissues. PMID:17967857

  18. Salmonella enterica ssp. arizonae infection in a 43-year-old Italian man with hypoglobulinemia: a case report and review of the literature

    Directory of Open Access Journals (Sweden)

    Noto Pasquale

    2011-07-01

    Full Text Available Abstract Introduction Salmonella enterica ssp. arizonae is an uncommon human pathogen with serious infections reported in immunocompromised hosts. In Europe, only a few cases have been described. Patients with this infection usually have a history of contact with reptiles or travel abroad. We present a case report of infection in a patient with hypoglobulinemia and a literature review. Case presentation We describe the case of a 43-year-old Caucasian Italian man with hypoglobulinemia who presented to our hospital with sepsis and diarrhea. A stool culture yielded S. enterica ssp. arizonae. Our patient was treated with oral ciprofloxacin and made a full recovery. We also present a review of the cases of S. enterica ssp. arizonae infections previously reported in Europe. Conclusions The majority of infections from S. enterica ssp. arizonae occur in patients who are immunocompromised. Data from the literature suggests that it may be difficult to eradicate the bacteria and thus, prolonged antibiotic courses are often used. It would be advisable for clinicians to investigate for pre-existing immune dysfunction if S. enterica ssp. arizonae is isolated. In Italy, although there have only been a few cases, the likely route of transmission remains unclear and requires further surveillance.

  19. Antibiotic resistance and diversity of Salmonella enterica serovars associated with broiler chickens.

    Science.gov (United States)

    Diarra, Moussa Sory; Delaquis, Pascal; Rempel, Heidi; Bach, Susan; Harlton, Colleen; Aslam, Mueen; Pritchard, Jane; Topp, Edward

    2014-01-01

    The objective of this study was to analyze the antibiotic resistance phenotype and genotype of Salmonella isolated from broiler production facilities. A total of 193 Salmonella isolates recovered from commercial farms in British Columbia, Canada, were evaluated. Susceptibility to antibiotics was determined with the Sensititre system. Virulence and antibiotic resistance genes were detected by PCR assay. Genetic diversity was determined by pulse-field gel electrophoresis (PFGE) typing. Seventeen serovars of Salmonella were identified. The most prevalent Salmonella serovars were Kentucky (29.0% of isolates), Typhimurium (23.8%), Enteritidis (13.5%), and Hadar (11.9%); serovars Heidelberg, Brandenburg, and Thompson were identified in 7.7, 4.1, and 3.6% of isolates, respectively. More than 43% of the isolates were simultaneously resistant to ampicillin, amoxicillin-clavulanic acid, ceftiofur, cefoxitim, and ceftriaxone. This β-lactam resistance pattern was observed in 33 (58.9%) of the Salmonella Kentucky isolates; 2 of these isolates were also resistant to chloramphenicol, streptomycin, sulfisoxazole, and tetracycline. Genes associated with resistance to aminoglycosides (aadA1, aadA2, and strA), β-lactams (blaCMY-2, blaSHV, and blaTEM), tetracycline (tetA and tetB), and sulfonamide (sul1) were detected among corresponding resistant isolates. The invasin gene (invA) and the Salmonella plasmid virulence gene (spvC) were found in 97.9 and 25.9% of the isolates, respectively, with 33 (71.7%) of the 46 Salmonella Typhimurium isolates and 17 (65.4%) of the 26 Salmonella Enteritidis isolates carrying both invA and spvC. PGFE typing revealed that the antibiotic-resistant serovars were genetically diverse. These data confirm that broiler chickens can be colonized by genetically diverse antibiotic-resistant Salmonella isolates harboring virulence determinants. The presence of such strains is highly relevant to food safety and public health.

  20. Antimicrobial Susceptibility and Plasmid Replicon Typing of Salmonella enterica serovar Kentucky isolates recovered from Broilers

    Science.gov (United States)

    Salmonella Kentucky has become the predominate serotype recovered from broiler slaughter in the United States and the prevalence of antimicrobial resistance (AMR) has increased dramatically in this serotype. Relationships between AMR, genotype, and plasmid replicon types were characterized for 600 ...

  1. Salmonella enterica Serovar Typhimurium-Dependent Regulation of Inducible Nitric Oxide Synthase Expression in Macrophages by Invasins SipB, SipC, and SipD and Effector SopE2

    OpenAIRE

    Cherayil, Bobby J.; McCormick, Beth A.; Bosley, Jacob

    2000-01-01

    When Salmonella enterica invades mammalian cells, it activates signals leading to increased expression of inflammatory mediators. One such mediator is nitric oxide (NO), which is produced under control of the enzyme inducible NO synthase (iNOS). Induction of iNOS in response to Salmonella infection has been demonstrated, but the bacterial effector molecules that regulate expression of the enzyme have not been identified. In the study reported here, an analysis of Salmonella-dependent iNOS exp...

  2. Colistin Resistance in Escherichia coli and Salmonella enterica Strains Isolated from Swine in Brazil

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    Adriano Savoia Morales

    2012-01-01

    Full Text Available Reports about acquired resistance to colistin in different bacteria species are increasing, including E. coli of animal origin, but reports of resistance in wild S. enterica of different serotypes from swine are not found in the literature. Results obtained with one hundred and twenty-six E. coli strains from diseased swine and one hundred and twenty-four S. enterica strains from diseased and carrier swine showed a frequency of 6.3% and 21% of colistin-resistant strains, respectively. When comparing the disk diffusion test with the agar dilution test to evaluate the strains, it was confirmed that the disk diffusion test is not recommended to evaluate colistin resistance as described previously. The colistin MIC 90 and MIC 50 values obtained to E. coli were 0.25 μg/mL and 0.5 μg/mL, the MIC 90 and MIC 50 to S. enterica were 1 μg/mL and 8 μg/mL. Considering the importance of colistin in control of nosocomial human infections with Gram-negative multiresistant bacteria, and the large use of this drug in animal production, the colistin resistance prevalence in enterobacteriaceae of animal origin must be monitored more closely.

  3. Herd problems and occupational zoonoses of Salmonella enterica serovars Typhimurium and Enteritidis infection in diarrheic cattle and buffalo calves in Egypt

    Directory of Open Access Journals (Sweden)

    Ahmed I. Youssef

    2012-12-01

    Full Text Available This study aimed to investigate Salmonella enterica serovar Typhimurium (ST and Salmonella enterica serovar Enteritidis (SEinfections of diarrheic calves in bovine dairy herds and their farm workers. Antimicrobial drug resistance profile of the isolates was determinedby standard disc diffusion assay. Fecal swabs were collected from diarrheic calves aged from one week to three months (Median age 33.5 days;from three farms representing cattle, native breed buffalo and cross-breed buffalo farms. Each farm was visited every three months for one year,from March 2011 to February 2012. Collected Swabs were examined by conventional bacterial culture and serotyped by multiplex PCR. Resultsshowed that Salmonella was isolated from 42/255 (18.66% diarrheic calves and 27/35 (77.14% calves died from diarrhea. Incidence of STwas significantly higher in calves died from diarrhea than diarrheic calves (P>0.001. On farm level, isolation rate of Salmonella was significantlyhigher in dead calves in cross-breed buffalo farm than native breed buffalo and cattle farms (P>0.001. On other hand, the incidence ofSalmonella among stool swabs of farm workers caring the calves was 8.13%; with isolation rate of (2.32% ST, (1.16% SE and (4.65% otherSalmonella serovas. Isolates showed a high percentage of multi-drug resistant profile (63.88%. All isolates were completely resistant to oxytetracycline,danofloxacin and oxacillin. Enrofloxacin was the drug of choice. In conclusion, a high incidence of mutli-drug resistant Salmonella infections was detected in cattle and buffalo calves that might be associated with high mortality rate in calves and zoonotic potential to the farmworkers. Adequate hygienic measures, proper immunization programs and regular training to the labor should be applied.

  4. Evaluation of corn oil as an additive in the pre-enrichment step to increase recovery of Salmonella enterica from oregano.

    Science.gov (United States)

    Jean-Gilles Beaubrun, Junia; Flamer, Marie-Laure; Addy, Nicole; Ewing, Laura; Gopinath, Gopal; Jarvis, Karen; Grim, Chris; Hanes, Darcy E

    2016-08-01

    Phenolic compounds associated with essential oils of spices and herbs possess a variety of antioxidant and antimicrobial properties that interfere with Salmonella detection from fresh and dried products. Finding a compound to neutralize the effect of these antimicrobial compounds, while allowing Salmonella growth during pre-enrichment, is a crucial step in both traditional pathogen isolation and molecular detection from these foods. This study evaluated the effectiveness of corn oil as a component of the pre-enrichment broth to counteract antimicrobial compounds properties and increase the recovery of Salmonella from spices. Oregano samples artificially contaminated with Salmonella enterica were pre-enriched in modified Buffered Peptone Water (mBPW) supplemented with and without 2% (vol/vol) corn oil respectively. Samples were incubated overnight at 37 °C. The results showed that recovery of Salmonella from oregano samples was increased by ≥50% when pre-enriched with corn oil. Serovars were confirmed using a PCR serotyping method. In addition, shot-gun metagenomics analyses demonstrated bacterial diversity and the effect of corn oil on the relative prevalence of Salmonella in the oregano samples. Modifying pre-enrichment broths with corn oil improved the detection and isolation of Salmonella from oregano, and may provide an alternative method for pathogen detection in dried food matrices such as spices. PMID:27052719

  5. Evaluation of corn oil as an additive in the pre-enrichment step to increase recovery of Salmonella enterica from oregano.

    Science.gov (United States)

    Jean-Gilles Beaubrun, Junia; Flamer, Marie-Laure; Addy, Nicole; Ewing, Laura; Gopinath, Gopal; Jarvis, Karen; Grim, Chris; Hanes, Darcy E

    2016-08-01

    Phenolic compounds associated with essential oils of spices and herbs possess a variety of antioxidant and antimicrobial properties that interfere with Salmonella detection from fresh and dried products. Finding a compound to neutralize the effect of these antimicrobial compounds, while allowing Salmonella growth during pre-enrichment, is a crucial step in both traditional pathogen isolation and molecular detection from these foods. This study evaluated the effectiveness of corn oil as a component of the pre-enrichment broth to counteract antimicrobial compounds properties and increase the recovery of Salmonella from spices. Oregano samples artificially contaminated with Salmonella enterica were pre-enriched in modified Buffered Peptone Water (mBPW) supplemented with and without 2% (vol/vol) corn oil respectively. Samples were incubated overnight at 37 °C. The results showed that recovery of Salmonella from oregano samples was increased by ≥50% when pre-enriched with corn oil. Serovars were confirmed using a PCR serotyping method. In addition, shot-gun metagenomics analyses demonstrated bacterial diversity and the effect of corn oil on the relative prevalence of Salmonella in the oregano samples. Modifying pre-enrichment broths with corn oil improved the detection and isolation of Salmonella from oregano, and may provide an alternative method for pathogen detection in dried food matrices such as spices.

  6. Characteristics of Salmonella enterica serovar 4,[5],12:i:- as a monophasic variant of serovar Typhimurium.

    Science.gov (United States)

    Ido, Noriko; Lee, Ken-ichi; Iwabuchi, Kaori; Izumiya, Hidemasa; Uchida, Ikuo; Kusumoto, Masahiro; Iwata, Taketoshi; Ohnishi, Makoto; Akiba, Masato

    2014-01-01

    Salmonella enterica subspecies enterica serovar 4,[5],12:i:- (S. 4,[5]12:i:-) is believed to be a monophasic variant of S. enterica serovar Typhimurium (S. Typhimurium). This study was conducted to corroborate this hypothesis and to identify the molecular and phenotypic characteristics of the S. 4,[5]12:i:- isolates in Japan. A total of 51 S. 4,[5]12:i:- isolates derived from humans, cattle, swine, chickens, birds, meat (pork), and river water in 15 prefectures in Japan between 2000 and 2010 were analyzed. All the S. 4,[5],12:i:- isolates were identified as S. Typhimurium by two different polymerase chain reactions (PCR) for identification of S. Typhimurium. Of the 51 S. 4,[5],12:i:- isolates, 39 (76.5%) harbored a 94-kb virulence plasmid, which is known to be specific for S. Typhimurium. These data suggest that the S. 4,[5],12:i:- isolates are monophasic variants of S. Typhimurium. The flagellar phase variation is induced by three adjacent genes (fljA, fljB, and hin) in the chromosome. The results of PCR mapping of this region and comparative genomic hybridization analysis suggested that the deletion of the fljAB operon and its flanking region was the major genetic basis of the monophasic phenotype of S. 4,[5],12:i:-. The fljAB operon and hin gene were detectable in eight of the S. 4,[5],12:i:- isolates with common amino acid substitutions of A46T in FljA and R140L in Hin. The introduction of these mutations into S. Typhimurium isolates led to the loss of selectability of isolates expressing the phase 2 H antigen. These data suggested that a point mutation was the genetic basis, at least in part, of the S. 4,[5],12:i:- isolates. The results of phenotypic analysis suggested that the S. 4,[5],12:i:- isolates in Japan consist of multiple distinct clones. This is the first detailed characterization of the S. 4,[5],12:i:- isolates derived from various sources across Japan.

  7. Characteristics of Salmonella enterica serovar 4,[5],12:i:- as a monophasic variant of serovar Typhimurium.

    Directory of Open Access Journals (Sweden)

    Noriko Ido

    Full Text Available Salmonella enterica subspecies enterica serovar 4,[5],12:i:- (S. 4,[5]12:i:- is believed to be a monophasic variant of S. enterica serovar Typhimurium (S. Typhimurium. This study was conducted to corroborate this hypothesis and to identify the molecular and phenotypic characteristics of the S. 4,[5]12:i:- isolates in Japan. A total of 51 S. 4,[5]12:i:- isolates derived from humans, cattle, swine, chickens, birds, meat (pork, and river water in 15 prefectures in Japan between 2000 and 2010 were analyzed. All the S. 4,[5],12:i:- isolates were identified as S. Typhimurium by two different polymerase chain reactions (PCR for identification of S. Typhimurium. Of the 51 S. 4,[5],12:i:- isolates, 39 (76.5% harbored a 94-kb virulence plasmid, which is known to be specific for S. Typhimurium. These data suggest that the S. 4,[5],12:i:- isolates are monophasic variants of S. Typhimurium. The flagellar phase variation is induced by three adjacent genes (fljA, fljB, and hin in the chromosome. The results of PCR mapping of this region and comparative genomic hybridization analysis suggested that the deletion of the fljAB operon and its flanking region was the major genetic basis of the monophasic phenotype of S. 4,[5],12:i:-. The fljAB operon and hin gene were detectable in eight of the S. 4,[5],12:i:- isolates with common amino acid substitutions of A46T in FljA and R140L in Hin. The introduction of these mutations into S. Typhimurium isolates led to the loss of selectability of isolates expressing the phase 2 H antigen. These data suggested that a point mutation was the genetic basis, at least in part, of the S. 4,[5],12:i:- isolates. The results of phenotypic analysis suggested that the S. 4,[5],12:i:- isolates in Japan consist of multiple distinct clones. This is the first detailed characterization of the S. 4,[5],12:i:- isolates derived from various sources across Japan.

  8. The outer membrane protease PgtE of Salmonella enterica interferes with the alternative complement pathway by cleaving factors B and H

    Directory of Open Access Journals (Sweden)

    Rauna eRiva

    2015-02-01

    Full Text Available The virulence factor PgtE is an outer membrane protease (omptin of the zoonotic pathogen Salmonella enterica that causes diseases ranging from gastroenteritis to severe enteric fever. It is surface exposed in bacteria that have a short-chain, i.e. rough LPS, as observed e.g. in bacteria residing inside macrophages or just emerging from them. We investigated whether PgtE cleaves the complement factors B (B and H (H, key proteins controlling formation and inactivation of the complement protein C3b and thereby the activity of the complement system. S. enterica serovar Typhimurium or omptin-expressing recombinant E. coli bacteria were incubated with purified human complement proteins or recombinant H fragments. PgtE cleaved both B and H, whereas its close homolog Pla of Yersinia pestis cleaved only H. H was cleaved at both N- and C-termini, while the central region resisted proteolysis. Because of multiple effects of PgtE on complement components (cleavage of C3, C3b, B and H we assessed its effect on the opsonophagocytosis of Salmonella. In human serum, C3 cleavage was dependent on proteolytically active PgtE. Human neutrophils interacted less with serum-opsonized FITC-stained S. enterica 14028R than with the isogenic ΔpgtE strain, as analyzed by flow cytometry. In conclusion, cleavage of B and H by PgtE, together with C3 cleavage, affects the C3-mediated recognition of S. enterica by human neutrophils, thus thwarting the immune protection against Salmonella.

  9. Activation and Genetic Modification of Human Monocyte-Derived Dendritic Cells using Attenuated Salmonella typhimurium

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    Agnieszka Michael

    2010-01-01

    Full Text Available Live attenuated bacterial vectors, such as Salmonella typhimurium, have shown promise as delivery vehicles for DNA. We have examined two new strains of S. typhimurium and their impact on dendritic cell maturation (CD12-sifA/aroC mutant and WT05-ssaV/aroC, both in TML background. Strain WT05 matured dendritic cells in a more efficient way; caused higher release of cytokines TNF-α, IL-12, IL-1β; and was efficient for gene transfer. These findings suggest that the genetic background of the attenuation can influence the pattern of inflammatory immune response to Salmonella infection.

  10. Antimicrobial resistance and virulence-associated genes of Salmonella enterica subsp. enterica serotypes Muenster, Florian, Omuna, and Noya strains isolated from clinically diarrheic humans in Egypt.

    Science.gov (United States)

    Osman, Kamelia M; Marouf, Sherif H; Alatfeehy, Nayerah

    2013-10-01

    Four serotypes recovered from clinically diarrheic human faecal samples (Salmonella Muenster, Salmonella Florian, Salmonella Omuna and Salmonella Noya) were investigated for the presence of 11 virulence genes (invA, avrA, ssaQ, mgtC, siiD, sopB, gipA, sodC1, sopE1, spvC, and bcfC) and their association with antibiotic resistance. The 4 Salmonella serotypes lacked virulence genes gipA and spvC. Resistance to 7 of the 14 antimicrobials was detected. The frequency of resistance, to lincomycin and streptomycin (100% of the Salmonella Muenster [2/5], Salmonella Florian [1/5], Salmonella Omuna [1/5], and Salmonella Noya [1/5] isolates), chloramphenicol (100% of the Salmonella Muenster [2/5] and Salmonella Florian [1/5] isolates) and trimethoprim-sulfamethoxazole (100% of the Salmonella Florian [1/5] and Salmonella Omuna [1/5] isolates) was an outstanding feature. With the rest of the antibiotics, the four Salmonella serotypes exhibited a great diversity in their resistance patterns. Overall, the four Salmonella serotypes were resistant to more than one antimicrobial. The antimicrobials to which the Salmonella Muenster, Salmonella Florian, and Salmonella Omuna isolates were resistant, contributed to five different antimicrobial resistance profiles. The virulence associated genes invA, ssaQ, siiD, sopB, and bcfC genes were 100% associated with certain antimicrobial resistance phenotypes (streptomycin and lincosamide) not recorded previously, and secondly, the presence of invA, avrA, ssaQ, mgtC, siiD, sopB, and bcfC was associated with resistance to chloramphenicol. The results of this study will help in understanding the spread of virulence genotypes and antibiotic resistance in Salmonella in the region of study.

  11. Crosstalk between virulence loci: regulation of Salmonella enterica pathogenicity island 1 (SPI-1) by products of the std fimbrial operon.

    Science.gov (United States)

    López-Garrido, Javier; Casadesús, Josep

    2012-01-01

    Invasion of intestinal epithelial cells is a critical step in Salmonella infection and requires the expression of genes located in Salmonella pathogenicity island 1 (SPI-1). A key factor for SPI-1 expression is DNA adenine (Dam) methylation, which activates synthesis of the SPI-1 transcriptional activator HilD. Dam-dependent regulation of hilD is postranscriptional (and therefore indirect), indicating the involvement of unknown cell functions under Dam methylation control. A genetic screen has identified the std fimbrial operon as the missing link between Dam methylation and SPI-1. We show that all genes in the std operon are part of a single transcriptional unit, and describe three previously uncharacterized ORFs (renamed stdD, stdE, and stdF). We present evidence that two such loci (stdE and stdF) are involved in Dam-dependent control of Salmonella SPI-1: in a Dam(-) background, deletion of stdE or stdF suppresses SPI-1 repression; in a Dam(+) background, constitutive expression of StdE and/or StdF represses SPI-1. Repression of SPI-1 by products of std operon explains the invasion defect of Salmonella Dam(-) mutants, which constitutively express the std operon. Dam-dependent repression of std in the ileum may be required to permit invasion, as indicated by two observations: constitutive expression of StdE and StdF reduces invasion of epithelial cells in vitro (1,000 fold) and attenuates Salmonella virulence in the mouse model (>60 fold). In turn, crosstalk between std and SPI-1 may play a role in intestinal infections by preventing expression of SPI-1 in the caecum, an intestinal compartment in which the std operon is known to be expressed.

  12. Reversion to virulence evaluation of a 9R vaccine strain of Salmonella enterica serovar gallinarum in commercial brown layers

    Directory of Open Access Journals (Sweden)

    AS Okamoto

    2010-03-01

    Full Text Available The live vaccine Cevac S. Gallinarum, made from a rough strain of Salmonella enterica subspecies enterica serotype Gallinarum is used for preventing fowl typhoid, a disease that still causes considerable economic losses in countries with a developing poultry industry. The objective of this paper was to evaluate a possible reversion to virulence of the strain used in a vaccine in commercial brown layers. Only Salmonella-free chicks were utilized. One hundred twenty (120 12-day-old Dekalb brown layers divided in two trials were used. The first trial had six groups of 15 birds each. Birds of group 1 were vaccinated with 10 doses of Cevac S. Gallinarum subcutaneously and 10 doses orally, in a total of 20 doses of vaccine. Then the birds of groups 2, 3, 4, and 5 received inocula that contained feces and a pool of organs with fragments of liver, heart, spleen, and cecal tonsils obtained from the immediately previous group. The second trial had three groups with 10 birds each. Birds in group 7 received inocula containing a pool of organs from birds of group 5 from trial 1, whilst the birds in group 8 were vaccinated subcutaneously with one dose of vaccine. Both trials included negative control groups (6 and 9. Throughout the experimental period, birds were monitored for reactions to the vaccination on the site of administration, clinical signs, and post-mortem lesions. In each passage, in addition to the birds euthanized to provide the inocula material, two birds from each group were euthanized for assessment of possible lesions, and their organs (liver, heart, spleen and cecal tonsils were cultured in an attempt to isolate the vaccine strain. Except for one bird from group 1, that had a local reaction on the site of vaccination - a small vesicle with less that 0.5 mm that persisted until the third day post vaccination -, no other bird had any local reaction to the vaccine or any visible clinical alteration. Birds in group 8 did not present any

  13. 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

    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...... findings here. Results Until 1997, DT104 infections made up less than 1 percent of all human salmonella infections. The strain isolated from patients in the first community outbreak of DT104 in Denmark, in 1998, was resistant to nalidixic acid and had reduced susceptibility to fluoroquinolones. The...... fluoroquinolones. Conclusions Our investigation of an outbreak of DT104 documented the spread of quinolone-resistant bacteria from food animals to humans; this spread was associated with infections that were difficult to treat. Because of the increase in quinolone resistance in salmonella, the use of...

  14. Genes ycfR, sirA and yigG contribute to the surface attachment of Salmonella enterica Typhimurium and Saintpaul to fresh produce.

    Directory of Open Access Journals (Sweden)

    Joelle K Salazar

    Full Text Available Salmonella enterica is a frequent contaminant of minimally-processed fresh produce linked to major foodborne disease outbreaks. The molecular mechanisms underlying the association of this enteric pathogen with fresh produce remain largely unexplored. In our recent study, we showed that the expression of a putative stress regulatory gene, ycfR, was significantly induced in S. enterica upon exposure to chlorine treatment, a common industrial practice for washing and decontaminating fresh produce during minimal processing. Two additional genes, sirA involved in S. enterica biofilm formation and yigG of unknown function, were also found to be differentially regulated under chlorine stress. To further characterize the roles of ycfR, sirA, and yigG in S. enterica attachment and survival on fresh produce, we constructed in-frame deletions of all three genes in two different S. enterica serovars, Typhimurium and Saintpaul, which have been implicated in previous disease outbreaks linked to fresh produce. Bacterial attachment to glass and polystyrene microtiter plates, cell aggregation and hydrophobicity, chlorine resistance, and surface attachment to intact spinach leaf and grape tomato were compared among wild-type strains, single-gene deletion mutants, and their respective complementation mutants. The results showed that deletions of ycfR, sirA, and yigG reduced bacterial attachment to glass and polystyrene as well as fresh produce surface with or without chlorine treatment in both Typhimurium and Saintpaul. Deletion of ycfR in Typhimurium significantly reduced bacterial chlorine resistance and the attachment to the plant surfaces after chlorinated water washes. Deletions of ycfR in Typhimurium and yigG in Saintpaul resulted in significant increase in cell aggregation. Our findings suggest that ycfR, sirA, and yigG collectively contribute to S. enterica surface attachment and survival during post-harvest minimal processing of fresh produce.

  15. Prevalence of subclinical Salmonella enterica infection in Danish finishing pig herds

    DEFF Research Database (Denmark)

    Stege, H.; Christensen, Julia; Nielsen, J.P.;

    2000-01-01

    included in the study. The representativeness of the selected herds was assessed, based on descriptive statistics of herd size and type. Totals of 1330 pen samples and 6814 blood samples were examined. The results from the meat-juice screening in the Danish S. enterica Control Programme were available...... of approximately 30-40 samples in 6 months). Ten herds had one or more positive meat-juice samples but were otherwise negative. S. Typhimurium was isolated from 30 of the 39 high-seroprevalence herds. Our conclusions were: (1) The within-herd seroprevalence among the 96 randomly selected Danish pig herds was low...

  16. [Isolation of a highly purified capsular polysaccharide from Salmonella enterica serovar Typhi (Salmonella typhi)--Vi-antigen and its use in serological diagnosis of typhoid fever].

    Science.gov (United States)

    Tesheva, A M; Aparin, P G; L'vov, V L

    2002-01-01

    For use in differential diagnostics of typhoid fever, samples of the capsular polysaccharide from Salmonella enterica serovar Typhi (usually named Vi-antigen) were isolated and characterized by physicochemical and serological methods. It was shown that only the sample of Vi-antigen with the minimal (0.57%) admixture of the corresponding lipopolysaccharide (LPS) from S. typhi retained a high serological activity in the tests with monoreceptor anti-Vi sera. However, it exhibited a substantially weaker reaction with sera from normal donors and patients with acute nontyphoid salmonelloses, than Vi-antigen preparations with a higher (0.8-1.2%) LPS content. The chromatographically pure Vi-antigen was purified by triple reprecipitation with hexadecyltrimethylammonium bromide. The content of the LPS admixture in the resulting Vi-antigen samples was quantitatively determined by GC. A high purification level of the Vi-antigen from the LPS admixture allows us to hope that this preparation could serve as a basic component of the test system for the diagnostics of typhoid fever. The English version of the paper.

  17. 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

    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...... 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...

  18. Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India

    Directory of Open Access Journals (Sweden)

    Irfan Ahmad Mir

    2015-07-01

    Conclusions: Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in prophylaxis and treatment regimen. This observation draws serious attention as poultry serves as an important source of transmission of these multidrug resistant Salmonella serovars to humans.

  19. Environmental contamination and detection of Salmonella enterica serovar enteritidis in laying flocks.

    Science.gov (United States)

    Davies, R; Breslin, M

    2001-12-01

    Faecal, dust and other environmental samples were collected from the floors, droppings belts, egg-collection systems and other areas of 14 cage-layer flocks, 10 barn egg production flocks and seven free-range flocks, and cultured for Salmonella species. The distribution of the organism varied with its prevalence and with the vaccination status of the birds. No one sample type was found to be suitable for identifying all contaminated houses. Salmonella was also frequently found on egg-packing equipment and in samples from rodents and wild birds. PMID:11787781

  20. 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

    trickle infected with low or moderate dose levels of Oesophagostomum spp. and challenge infected with S. Typhimurium. In another experiment, pigs were inoculated with S. Typhimurium followed by a challenge exposure to either Oesophagostomum, Trichuris or Ascaris. Enhancement of the Salmonella infection......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...