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Sample records for salmonella typhi gene

  1. In vivo expression of Salmonella enterica serotype Typhi genes in the blood of patients with typhoid fever in Bangladesh.

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    Alaullah Sheikh

    2011-12-01

    Full Text Available Salmonella enterica serotype Typhi is the cause of typhoid fever. It is a human-restricted pathogen, and few data exist on S. Typhi gene expression in humans.We applied an RNA capture and amplification technique, Selective Capture of Transcribed Sequences (SCOTS, and microarray hybridization to identify S. Typhi transcripts expressed in the blood of five humans infected with S. Typhi in Bangladesh. In total, we detected the expression of mRNAs for 2,046 S. Typhi genes (44% of the S. Typhi genome in human blood; expression of 912 genes was detected in all 5 patients, and expression of 1,100 genes was detected in 4 or more patients. Identified transcripts were associated with the virulence-associated PhoP regulon, Salmonella pathogenicity islands, the use of alternative carbon and energy sources, synthesis and transport of iron, thiamine, and biotin, and resistance to antimicrobial peptides and oxidative stress. The most highly represented group were genes currently annotated as encoding proteins designated as hypothetical, unknown, or unclassified. Of the 2,046 detected transcripts, 1,320 (29% of the S. Typhi genome had significantly different levels of detection in human blood compared to in vitro cultures; detection of 141 transcripts was significantly different in all 5 patients, and detection of 331 transcripts varied in at least 4 patients. These mRNAs encode proteins of unknown function, those involved in energy metabolism, transport and binding, cell envelope, cellular processes, and pathogenesis. We confirmed increased expression of a subset of identified mRNAs by quantitative-PCR.We report the first characterization of bacterial transcriptional profiles in the blood of patients with typhoid fever. S. Typhi is an important global pathogen whose restricted host range has greatly inhibited laboratory studies. Our results suggest that S. Typhi uses a largely uncharacterized genetic repertoire to survive within cells and utilize alternate

  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. Antimicrobial susceptibility patterns of Salmonella typhi and ...

    African Journals Online (AJOL)

    The aim of this study was to determine antimicrobial susceptibility testing patterns of Candida Albicans and Salmonella typhi isolates. Fifteen isolates of each microorganism were collected from three hospitals located in Dar es Salaam region within a 3-month period in the year 2005. Candida Albicans and Salmonella typhi ...

  4. Quinolone resistance in Salmonella enterica serovar Typhi ...

    African Journals Online (AJOL)

    over Asia and Africa) emerged from Southeast Asia and then spread to other regions of the world [13]. Travellers also played a significant role in spreading the resistant. Salmonella Typhi, especially to the developed world [16,. 17]. The quinolone-resistant Salmonella Typhi is not only prevalent in hospital settings but also in ...

  5. Identification and characterization of Salmonella typhi isolates from Southwest Sumba District, East Nusa Tenggara based on 16S rRNA gene sequences

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    CHARIS AMARANTINI

    2011-01-01

    Full Text Available Amarantini C, Sembiring L, Kushadiwijaya H, Asmara W (2011 Identification and characterization of Salmonella typhi isolates from Southwest Sumba District, East Nusa Tenggara based on 16S rRNA gene sequences. Biodiversitas 12: 1-6. The incidence rate of typhoid fever in the Southwest Sumba District, East Nusa Tenggara was approximately about 725/100,000. In spite of such rate, there was not much known-yet about the molecular epidemiology of the disease. Thus, having accurate data and a strong discriminatory ability was crucial to scrutinize the molecular epidemiology of S. typhi with a molecular phylogenetic approach based on 16S rRNA gene sequences. Sixteen isolates representative of S. typhi from different geographical regions in Southwest Sumba District along with the reference strain S. typhi NCTC 786 had been identified and characterised based on 16S rRNA gene sequences using PCR amplification and sequencing. The 16S rRNA sequences data were aligned with the corresponding available S. typhi sequences retrieved from the NCBI database by using CLUSTAL X software. Phylogenetic trees were generated with PHYLIP software package. Molecular phylogenetic analysis indicated that all the isolates belong to S. typhi species were suggested by their relativity with the type strain of S. typhi ATCC19430T. It was also found that the isolates which belong to S. typhi species formed several different centers of diversity within the 16S rRNA gene tree. Each clade consisted of the strains from different geographical places in the District. Thus, to conclude the inquiry, there was evident inter-geographical spread of the strains and it tended to spread further into more remote areas in the District.

  6. Salmonella typhi time to change empiric treatment

    DEFF Research Database (Denmark)

    Gade, C.; Engberg, J.; Weis, N.

    2008-01-01

    In the present case series report we describe seven recent cases of typhoid fever. All the patients were travellers returning from Pakistan, where typhoid is endemic. Salmonella typhi isolated from the patients by blood culture were reported as intermediary susceptible to fluoroquinolones in six...

  7. Quinolone resistance in Salmonella enterica serovar Typhi ...

    African Journals Online (AJOL)

    Introduction. The human restricted bacteria, Salmonella enterica serovar. Typhi is the major cause of typhoid fever (or enteric fever), a characteristic severe systemic illness [1]. In 2010, typhoid fever accounted for an estimated global burden of. 27 million new cases and 200,000 deaths [2]. For over two decades, S. enterica ...

  8. Salmonella typhi in the democratic republic of the congo: fluoroquinolone decreased susceptibility on the rise.

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    Octavie Lunguya

    Full Text Available BACKGROUND: Drug resistance of Salmonella enterica serovar Typhi (Salmonella Typhi to first-line antibiotics is emerging in Central Africa. Although increased use of fluoroquinolones is associated with spread of resistance, Salmonella Typhi with decreased ciprofloxacin susceptibility (DCS has rarely been reported in Central Africa. METHODOLOGY/PRINCIPAL FINDINGS: As part of a microbiological surveillance study in the Democratic Republic of the Congo (DR Congo, Salmonella Typhi isolates from bloodstream infections were collected prospectively between 2007 and 2011. The genetic relationship of the Salmonella Typhi isolates was assessed by pulsed-field gel electrophoresis (PFGE. The antimicrobial resistance profile of the isolates was determined and mutations associated with DCS were studied. In total, 201 Salmonella Typhi isolates were collected. More than half of the Salmonella Typhi isolates originated from children and young adults aged 5-19. Thirty different PFGE profiles were identified, with 72% of the isolates showing a single profile. Multidrug resistance, DCS and azithromycin resistance were 30.3%, 15.4% and 1.0%, respectively. DCS was associated with point mutations in the gyrA gene at codons 83 and 87. CONCLUSIONS/SIGNIFICANCE: Our study describes the first report of widespread multidrug resistance and DCS among Salmonella Typhi isolates from DR Congo. Our findings highlight the need for increased microbiological diagnosis and surveillance in DR Congo, being a prerequisite for rational use of antimicrobials and the development of standard treatment guidelines.

  9. Ludwig's angina by Salmonella Typhi: a clinical dilemma.

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    Mahajan, R K; Sharma, S; Madan, P; Sharma, N

    2015-01-01

    Salmonella Typhi has rarely been associated with focal abscesses; and in literature, there is no evidence of its association with abscesses in the neck spaces. Ability of Salmonella Typhi to invade and localise in the neck spaces not only poses a diagnostic challenge but also underscores the necessity to understand the mechanisms that facilitate Salmonella Typhi to establish infections at sites completely non-traditional to the organism.

  10. Ludwig′s angina by Salmonella Typhi: A clinical dilemma

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    R K Mahajan

    2015-01-01

    Full Text Available Salmonella Typhi has rarely been associated with focal abscesses; and in literature, there is no evidence of its association with abscesses in the neck spaces. Ability of Salmonella Typhi to invade and localise in the neck spaces not only poses a diagnostic challenge but also underscores the necessity to understand the mechanisms that facilitate Salmonella Typhi to establish infections at sites completely non-traditional to the organism.

  11. Salmonella paratyphi C: genetic divergence from Salmonella choleraesuis and pathogenic convergence with Salmonella typhi.

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    Wei-Qiao Liu

    Full Text Available BACKGROUND: Although over 1400 Salmonella serovars cause usually self-limited gastroenteritis in humans, a few, e.g., Salmonella typhi and S. paratyphi C, cause typhoid, a potentially fatal systemic infection. It is not known whether the typhoid agents have evolved from a common ancestor (by divergent processes or acquired similar pathogenic traits independently (by convergent processes. Comparison of different typhoid agents with non-typhoidal Salmonella lineages will provide excellent models for studies on how similar pathogens might have evolved. METHODOLOGIES/PRINCIPAL FINDINGS: We sequenced a strain of S. paratyphi C, RKS4594, and compared it with previously sequenced Salmonella strains. RKS4594 contains a chromosome of 4,833,080 bp and a plasmid of 55,414 bp. We predicted 4,640 intact coding sequences (4,578 in the chromosome and 62 in the plasmid and 152 pseudogenes (149 in the chromosome and 3 in the plasmid. RKS4594 shares as many as 4346 of the 4,640 genes with a strain of S. choleraesuis, which is primarily a swine pathogen, but only 4008 genes with another human-adapted typhoid agent, S. typhi. Comparison of 3691 genes shared by all six sequenced Salmonella strains placed S. paratyphi C and S. choleraesuis together at one end, and S. typhi at the opposite end, of the phylogenetic tree, demonstrating separate ancestries of the human-adapted typhoid agents. S. paratyphi C seemed to have suffered enormous selection pressures during its adaptation to man as suggested by the differential nucleotide substitutions and different sets of pseudogenes, between S. paratyphi C and S. choleraesuis. CONCLUSIONS: S. paratyphi C does not share a common ancestor with other human-adapted typhoid agents, supporting the convergent evolution model of the typhoid agents. S. paratyphi C has diverged from a common ancestor with S. choleraesuis by accumulating genomic novelty during adaptation to man.

  12. Physiological and Immunological Regulations in Caenorhabditis elegans Infected with Salmonella enterica serovar Typhi.

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    Sivamaruthi, Bhagavathi Sundaram; Balamurugan, Krishnaswamy

    2014-03-01

    Studies pertaining to Salmonella enterica serovar Typhimurium infection by utilizing model systems failed to mimic the essential aspects of immunity induced by Salmonella enterica serovar Typhi, as the determinants of innate immunity are distinct. The present study investigated the physiological and innate immune responses of S. Typhi infected Caenorhabditis elegans and also explored the Ty21a mediated immune enhancement in C. elegans. Ty21a is a known live vaccine for typhoidal infection in human beings. Physiological responses of C. elegans infected with S. Typhi assessed by survival and behavioral assays revealed that S. Typhi caused host mortality by persistent infection. However, Ty21a exposure to C. elegans was not harmful. Ty21a pre-exposed C. elegans, exhibited significant resistance against S. Typhi infection. Elevated accumulation of S. Typhi inside the infected host was observed when compared to Ty21a exposures. Transcript analysis of candidate innate immune gene (clec-60, clec-87, lys-7, ilys-3, scl-2, cpr-2, F08G5.6, atf-7, age-1, bec-1 and daf-16) regulations in the host during S. Typhi infection have been assessed through qPCR analysis to understand the activation of immune signaling pathways during S. Typhi infections. Gene silencing approaches confirmed that clec-60 and clec-87 has a major role in the defense system of C. elegans during S. Typhi infection. In conclusion, the study revealed that preconditioning of host with Ty21a protects against subsequent S. Typhi infection.

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

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    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.

  14. Behaviour of temperate phage Mu in Salmonella typhi.

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    Soberon, M; Gama, M J; Richelle, J; Martuscelli, J

    1986-01-01

    We have developed a convenient system for genetic analysis of Salmonella typhi exploiting the properties of the mutator phage Mu. In spite of the fact that wild-type Salmonella typhi strains do not allow Mu to form plaques on them, we have shown that these strains are actually sensitive to the phage. It proved possible to use Mu to induce mutations and to promote intra- and interspecific genetic transfer, without having to introduce the phage into the bacteria by means other than infection. Furthermore, we isolated Salmonella typhi derivatives on which Mu formed plaques, and studied the behaviour of Mu in these and wild-type strains.

  15. Effect of garlic ( Allium sativum ) on Salmonella typhi infection ...

    African Journals Online (AJOL)

    The effect of consumption of garlic (Allium sativum) in treating Salmonella typhi infection and on the gastrointestinal flora and hematological parameters of rats was investigated. Crude garlic extract inhibited the growth of S. typhi on agar plate with a zone of inhibition averaging 23.8 mm in diameter using the agar diffusion ...

  16. Overexpression of Salmonella enterica serovar Typhi recA gene confers fluoroquinolone resistance in Escherichia coli DH5α

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    M.A.M. Yassien

    2015-11-01

    Full Text Available A spontaneous fluoroquinolone-resistant mutant (STM1 was isolated from its parent Salmonella enterica serovar Typhi (S. Typhi clinical isolate. Unlike its parent isolate, this mutant has selective resistance to fluoroquinolones without any change in its sensitivity to various other antibiotics. DNA gyrase assays revealed that the fluoroquinolone resistance phenotype of the STM1 mutant did not result from alteration of the fluoroquinolone sensitivity of the DNA gyrase isolated from it. To study the mechanism of fluoroquinolone resistance, a genomic library from the STM1 mutant was constructed in Escherichia coli DH5α and two recombinant plasmids were obtained. Only one of these plasmids (STM1-A conferred the selective fluoroquinolone resistance phenotype to E. coli DH5α. The chromosomal insert from STM1-A, digested with EcoRI and HindIII restriction endonucleases, produced two DNA fragments and these were cloned separately into pUC19 thereby generating two new plasmids, STM1-A1 and STM1-A2. Only STM1-A1 conferred the selective fluoroquinolone resistance phenotype to E. coli DH5α. Sequence and subcloning analyses of STM1-A1 showed the presence of an intact RecA open reading frame. Unlike that of the wild-type E. coli DH5α, protein analysis of a crude STM1-A1 extract showed overexpression of a 40 kDa protein. Western blotting confirmed the 40 kDa protein band to be RecA. When a RecA PCR product was cloned into pGEM-T and introduced into E. coli DH5α, the STM1-A11 subclone retained fluoroquinolone resistance. These results suggest that overexpression of RecA causes selective fluoroquinolone resistance in E. coli DH5α.

  17. Exploring insights for virulent gene inhibition of multidrug resistant Salmonella typhi, Vibrio cholerae, and Staphylococcus areus by potential phytoligands via in silico screening.

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    Skariyachan, Sinosh; Jayaprakash, Nisha; Bharadwaj, Navya; Narayanappa, Rajeswari

    2014-01-01

    In our recent studies on prevalence of multidrug resistant pathogens in Byramangala reservoir, Karnataka, India, we identified Salmonella typhi, Staphylococcus aureus, and Vibrio cholerae which had acquired multiple drug resistance (MDR) and emerged as superbugs. Hence, there is a pressing demand to identify alternative therapeutic remedies. Our study focused on the screening of herbal leads by structure-based virtual screening. The virulent gene products of these pathogens towards Kanamycin(aph), Trimethoprim(dfrA1), Methicillin (mecI), and Vancomycin (vanH) were identified as the probable drug targets and their 3D structures were predicted by homology modeling. The predicted models showed good stereochemical validity. By extensive literature survey, we selected 58 phytoligands and their drug likeliness and pharmacokinetic properties were computationally predicted. The inhibitory properties of these ligands against drug targets were studied by molecular docking. Our studies revealed that Baicalein from S. baicalensis (baikal skullcap) and Luteolin from Taraxacum officinale (dandelion) were identified as potential inhibitors against aph of S. typhi. Resveratrol from Vitis vinifera (grape vine) and Wogonin from S. baicalensis were identified as potential inhibitors against dfrA1 of S. typhi. Herniarin from Herniaria glabra (rupture worts) and Pyrocide from Daucus carota (Carrot) were identified as the best leads against dfrA1 of V. cholerae. Taraxacin of T. officinale (weber) and Luteolin were identified as potential inhibitors against Mec1. Apigenin from Coffee arabica (coffee) and Luteolin were identified as the best leads against vanH of S. aureus. Our findings pave crucial insights for exploring alternative therapeutics against MDR pathogens.

  18. A Rare Case of Salmonella typhi Meningitis in an Eleven Month Old ...

    African Journals Online (AJOL)

    Non-typhoidal Salmonella are infrequent causes of childhood meningitis. Most reports of Salmonella typhi meningeal infections are confined to neonates. A rare instance of S. typhi in an otherwise healthy eleven month old infant is being reported. Keywords: Salmonella typhi, meningitis, infant.

  19. 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.

  20. The DNA static curvature has a role in the regulation of the ompS1 porin gene in Salmonella enterica serovar Typhi.

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    De la Cruz, Miguel Angel; Merino, Enrique; Oropeza, Ricardo; Téllez, Juan; Calva, Edmundo

    2009-07-01

    The DNA static curvature has been described to play a key role as a regulatory element in the transcription process of several bacterial genes. Here, the role of DNA curvature in the expression of the ompS1 porin gene in Salmonella enterica serovar Typhi is described. The web server mutacurve was used to predict mutations that diminished or restored the extent of DNA curvature in the 5' regulatory region of ompS1. Using these predictions, curvature was diminished by site-directed mutagenesis of only two residues, and curvature was restored by further mutagenesis of the same two residues. Lowering the extent of DNA curvature resulted in an increase in ompS1 expression and in the diminution of the affinity of the silencer proteins H-NS and StpA for the ompS1 5' regulatory region. These mutations were in a region shown not to contain the H-NS nucleation site, consistent with the notion that the effect on expression was due to changes in DNA structural topology.

  1. Salmonella enterica serovar Typhi and the pathogenesis of typhoid fever.

    Science.gov (United States)

    Dougan, Gordon; Baker, Stephen

    2014-01-01

    Salmonella enterica serovar Typhi, the cause of typhoid, is host restricted to humans. S. Typhi has a monophyletic population structure, indicating that typhoid in humans is a relatively new disease. Antimicrobial usage is reshaping the current S. Typhi global population and may be driving the emergence of a specific haplotype, H58, that is well adapted to transmission in modern settings and is able to resist antimicrobial killing more efficiently than other S. Typhi. Evidence gathered through genomics and functional studies using the mouse and in vitro cell systems, together with clinical investigations, has provided insight into the mechanisms that underpin the pathogenesis of human typhoid and host restriction. Here we review the latest scientific advances in typhoid research and discuss how these novel approaches are changing our understanding of the disease.

  2. Gangrene of the limb complicating Salmonella typhi Septicaemia in ...

    African Journals Online (AJOL)

    ... of the metatarsophalangeal joints two months after admission. This report is to alert clinicians about this rare complication of a common curable disease with a view to anticipating the possibility of it occurring as well as considering appropriate preventive measures. Keywords: Gangrene, foot, Salmonella typhi, septicaemia ...

  3. Detection of Salmonella typhi agglutinins in sera of patients with ...

    African Journals Online (AJOL)

    Annals of African Medicine. Introduction. Despite the introduction of several serological tests,[1-7] Widal test is commonly applied for diagnosis of typhoid fever. As the typhi antigen is shared by a large number of organisms from the Salmonella genus and other related organisms,[1,2,8] a positive. Widal test is therefore likely ...

  4. 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.

  5. Anaerobiosis induced virulence of Salmonella typhi

    DEFF Research Database (Denmark)

    Kapoor, Sarika; Singh, R D; Sharma, P C

    2002-01-01

    BACKGROUND & OBJECTIVES: Anaerobic conditions are frequently encountered by pathogens invading the gastrointestinal tract due to low/limiting oxygen conditions prevalent in the small intestine. This anaerobic stress has been suggested to enhance the virulence of gut pathogens. In the present stud...... dismutase (SOD) and catalase. INTERPRETATION & CONCLUSION: Our results suggest that exposure of S. Typhi to anaerobic conditions enhances its virulence....

  6. Identification and characterization of a gene stp17 located on the linear plasmid pBSSB1 as an enhanced gene of growth and motility in Salmonella enterica serovar Typhi

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    Haifang Zhang

    2016-10-01

    Full Text Available The linear plasmid pBSSB1 mediates the flagellar phase variation in H:z66 positive Salmonella enterica serovar Typhi (S. Typhi. The gene named stp17 (S. Typhi plasmid number 17 gene is located on pBSSB1 and encodes the protein STP17. The expression pattern at the protein-level and function of STP17 remains unknown. In this study, the recombinant protein STP17His6 was expressed, successfully purified and used to prepare the polyclonal anti-STP17 antibody. We detected protein-level expression of stp17 in S. Typhi and further investigated the protein expression characteristics of stp17 in different growth phases by western blot analysis. The effects of STP17 on bacterial growth and motility were analyzed. In addition, the structure of STP17 was predicted on line and the active site of STP17 was identified by site-directed mutagenesis. The results showed that STP17 was expressed stably in the wild type strain of S. Typhi. STP17 expression at the protein level peaks when cultures reach an OD600 value of 1.2. The growth rate and motility of the Δstp17 strain were significantly decreased compared with the wild type strain (P<0.05 and this phenotype was restored in the stp17 complementary strain. Moreover, the growth rate and motility of the stp17 over-expression strain was greater than the wild type strain. STP17 contains nine Helix segments, six Stand segments and some Coil segments in the secondary structural level. The top-ranked 3-D structure of STP17 predicted by I-TASSER contains a putative ATPase domain and the amino acid residues of GLY16, GLY19, LYS20, ASN133, LYS157 and LYS158 may be the active site residues of STP17. Finally, STP17 was able to catalyze the ATP to ADP reaction, suggesting that STP17 may be an ATPase. To our knowledge, this is the first report describing the protein expression characteristics of STP17 in S. Typhi, showing that STP17 promotes bacterial growth and motility, which may be associated with its potential ATPase

  7. Use of TaqMan® real-time PCR for rapid detection of Salmonella enterica serovar Typhi.

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    Ranjbar, Reza; Naghoni, Ali; Farshad, Shohreh; Lashini, Hadi; Najafi, Ali; Sadeghifard, Nourkhoda; Mammina, Caterina

    2014-06-01

    We evaluated the performances of a newly designed real-time polymerase chain reaction (PCR) assay using TaqMan® probes to detect Salmonella Typhi. TaqMan® real-time PCR assays were performed by designed primers and probe based on the staG gene for detecting S. Typhi. The specificity of the assay was evaluated on 15 Salmonella serovars. The analytical specificity was evaluated on 20 non-Salmonella microorganisms. The analytical sensitivity was assessed using decreasing DNA quantities of S. Typhi ATCC 19430. Finally the detection capability of the TaqMan® real-time PCR assay on isolates recovered from patients with Salmonella infections was compared to the conventional PCR assay. Only S. Typhi strain had positive results when subjected to the assay using Typhi-specific real-time PCR. No amplification products were observed in real-time PCR with any of the non-Salmonella microorganisms tested. The TaqMan® real-time PCR was more sensitive than the conventional PCR. In conclusion, we found that the easy-to-use real-time PCR assays were faster than conventional PCR systems. The staG-based TaqMan® real-time PCR assay showed to be specific and sensitive method for the safe and rapid detection of the S. Typhi.

  8. Antagonistic Activity of Lactobacillus Isolates against Salmonella typhi In Vitro

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    Abdel-Daim, Amira; Hassouna, Nadia; Hafez, Mohamed; Ashor, Mohamed Seif Aldeen; Aboulwafa, Mohammad M.

    2013-01-01

    Background. Enteric fever is a global health problem, and rapidly developing resistance to various drugs makes the situation more alarming. The potential use of Lactobacillus to control typhoid fever represents a promising approach, as it may exert protective actions through various mechanisms. Methods. In this study, the probiotic potential and antagonistic activities of 32 Lactobacillus isolates against Salmonella typhi were evaluated. The antimicrobial activity of cell free supernatants of Lactobacillus isolates, interference of Lactobacillus isolates with the Salmonella adherence and invasion, cytoprotective effect of Lactobacillus isolates, and possibility of concurrent use of tested Lactobacillus isolates and antibiotics were evaluated by testing their susceptibilities to antimicrobial agents, and their oxygen tolerance was also examined. Results. The results revealed that twelve Lactobacillus isolates could protect against Salmonella typhi infection through interference with both its growth and its virulence properties, such as adherence, invasion, and cytotoxicity. These Lactobacillus isolates exhibited MIC values for ciprofloxacin higher than those of Salmonella typhi and oxygen tolerance and were identified as Lactobacillus plantarum. Conclusion. The tested Lactobacillus plantarum isolates can be introduced as potential novel candidates that have to be subjected for in vivo and application studies for treatment and control of typhoid fever. PMID:24191248

  9. Vaccination of active component US military personnel against Salmonella Typhi.

    Science.gov (United States)

    Porter, Chad K; Sorrell, Tia; Mitra, Indrani; Riddle, Mark S

    2017-03-27

    Vaccination against Salmonella Typhi is one of the leading public health interventions reducing the risk of typhoid fever. There are two available licensed vaccines, Vivotif, oral live-attenuated, and Typhim Vi, intramuscular Vi capsular polysaccharide. The US military is a high risk travel population commonly vaccinated for S. Typhi. We describe the use of S. Typhi vaccination in this population and the acute reactogenicity profile of these vaccines. Data were obtained from the Defense Medical Surveillance System and vaccination identified between 1998 and 2011 from vaccination codes. Clinical outcomes were assessed for four weeks post vaccination. Adverse event rates and odds ratios were estimated across the two vaccine types. A total of 1.9million predominately male military personnel received 3.6 million S. Typhi vaccinations with 94.3% of vaccinees receiving the Vi capsule vaccine though variability in the vaccine administered was observed. Receipt of other vaccinations in the 6months surrounding the S. Typhi vaccine was common. Rates of nausea (195 per 100,000 vaccinations), headache (13 per 100,000 vaccinations) and fever (40 per 100,000 vaccinations) were significantly higher following Vi capsule vaccination compared to receipt of Vivotif (130, 2, 10 per 100,000 vaccinations, respectively). In contrast the rates of rash and non-infectious diarrhea (186 and 426 per 100,000 vaccinations, respectively) were increased in those receiving Vivotif compared to the Vi capsule vaccine. The US military is a major consumer of S. Typhi vaccines. The parenterally administered vaccine appears to be more amenable, though we were limited in our ability to assess the reasons for its higher usage. While we observed a higher rate of several adverse events in subjects receiving the intramuscular vaccination, the overall rate of these events was low. Future studies assessing more long-term health outcomes are warranted. Published by Elsevier Ltd.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Detection of Salmonella typhi utilizing bioconjugated fluorescent polymeric nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Swati, E-mail: swatijain.iitd@gmail.com; Chattopadhyay, Sruti, E-mail: sruticiitd@gmail.com; Jackeray, Richa; Abid, Zainul; Singh, Harpal, E-mail: harpal2000@yahoo.com [Centre for Biomedical Engineering, Indian Institute of Technology-Delhi (India)

    2016-05-15

    Present work demonstrates effective utilization of functionalized polymeric fluorescent nanoparticles as biosensing probe for the detection of Salmonella typhi bacteria on modified polycarbonate (PC) filters in about 3 h. Antibody modified-PC membranes were incubated with contaminated bacterial water for selective capturing which were detected by synthesized novel bioconjugate probe. Core–shell architecture of polymeric nanoparticles endows them with aqueous stabilization and keto-enolic functionalities making them usable for covalently linking S. typhi antibodies without any crosslinker or activator. Bradford analysis revealed that one nanoparticle has an average of 3.51 × 10{sup −19} g or 21 × 10{sup 4} bound S. typhi Ab molecules. Analysis of the regions of interest (ROI) in fluorescent micrographs of modified fluoroimmunoassay showed higher detection sensitivity of 5 × 10{sup 2} cells/mL due to signal amplification unlike conventional naked dye FITC-Ab conjugate. Fluorescence of pyrene dye remained same on immobilization of biomolecules and nanoparticles showed stable fluorescent intensity under prolong exposure to laser owing to protective polymeric layer allowing accurate identification of bacteria. Surface-functionalized PC matrix and fluorescent label NPs permit covalent interactions among biomolecules enhancing signal acquisitions showing higher detection efficiency as compared to conventional microtiter plate-based system. Our novel immunoassay has the potential to be explored as rapid detection method for identifying S. typhi contaminations in water.Graphical Abstract.

  12. Detection of Salmonella typhi utilizing bioconjugated fluorescent polymeric nanoparticles

    Science.gov (United States)

    Jain, Swati; Chattopadhyay, Sruti; Jackeray, Richa; Abid, Zainul; Singh, Harpal

    2016-05-01

    Present work demonstrates effective utilization of functionalized polymeric fluorescent nanoparticles as biosensing probe for the detection of Salmonella typhi bacteria on modified polycarbonate (PC) filters in about 3 h. Antibody modified-PC membranes were incubated with contaminated bacterial water for selective capturing which were detected by synthesized novel bioconjugate probe. Core-shell architecture of polymeric nanoparticles endows them with aqueous stabilization and keto-enolic functionalities making them usable for covalently linking S. typhi antibodies without any crosslinker or activator. Bradford analysis revealed that one nanoparticle has an average of 3.51 × 10-19 g or 21 × 104 bound S. typhi Ab molecules. Analysis of the regions of interest (ROI) in fluorescent micrographs of modified fluoroimmunoassay showed higher detection sensitivity of 5 × 102 cells/mL due to signal amplification unlike conventional naked dye FITC-Ab conjugate. Fluorescence of pyrene dye remained same on immobilization of biomolecules and nanoparticles showed stable fluorescent intensity under prolong exposure to laser owing to protective polymeric layer allowing accurate identification of bacteria. Surface-functionalized PC matrix and fluorescent label NPs permit covalent interactions among biomolecules enhancing signal acquisitions showing higher detection efficiency as compared to conventional microtiter plate-based system. Our novel immunoassay has the potential to be explored as rapid detection method for identifying S. typhi contaminations in water.

  13. Temporal fluctuation of multidrug resistant salmonella typhi haplotypes in the mekong river delta region of Vietnam.

    Directory of Open Access Journals (Sweden)

    Kathryn E Holt

    2011-01-01

    Full Text Available typhoid fever remains a public health problem in Vietnam, with a significant burden in the Mekong River delta region. Typhoid fever is caused by the bacterial pathogen Salmonella enterica serovar Typhi (S. Typhi, which is frequently multidrug resistant with reduced susceptibility to fluoroquinolone-based drugs, the first choice for the treatment of typhoid fever. We used a GoldenGate (Illumina assay to type 1,500 single nucleotide polymorphisms (SNPs and analyse the genetic variation of S. Typhi isolated from 267 typhoid fever patients in the Mekong delta region participating in a randomized trial conducted between 2004 and 2005.the population of S. Typhi circulating during the study was highly clonal, with 91% of isolates belonging to a single clonal complex of the S. Typhi H58 haplogroup. The patterns of disease were consistent with the presence of an endemic haplotype H58-C and a localised outbreak of S. Typhi haplotype H58-E2 in 2004. H58-E2-associated typhoid fever cases exhibited evidence of significant geo-spatial clustering along the Sông H u branch of the Mekong River. Multidrug resistance was common in the established clone H58-C but not in the outbreak clone H58-E2, however all H58 S. Typhi were nalidixic acid resistant and carried a Ser83Phe amino acid substitution in the gyrA gene.the H58 haplogroup dominates S. Typhi populations in other endemic areas, but the population described here was more homogeneous than previously examined populations, and the dominant clonal complex (H58-C, -E1, -E2 observed in this study has not been detected outside Vietnam. IncHI1 plasmid-bearing S. Typhi H58-C was endemic during the study period whilst H58-E2, which rarely carried the plasmid, was only transient, suggesting a selective advantage for the plasmid. These data add insight into the outbreak dynamics and local molecular epidemiology of S. Typhi in southern Vietnam.

  14. Molecular diagnosis of Salmonella typhi and its virulence in suspected typhoid blood samples through nested multiplex PCR.

    Science.gov (United States)

    Prabagaran, Solai Ramatchandirane; Kalaiselvi, Vellingiri; Chandramouleeswaran, Naganathan; Deepthi, Krishnan Nair Geetha; Brahmadathan, Kootallur Narayanan; Mani, Mariappa

    2017-08-01

    A nested multiplex polymerase chain reaction (PCR) based diagnosis was developed for the detection of virulent Salmonella typhi in the blood specimens from patients suspected for typhoid fever. After the Widal test, two pairs of primers were used for the detection of flagellin gene (fliC) of S. typhi. Among them, those positive for fliC alone were subjected to identification of genes in Via B operon of Salmonella Pathogenesity Island (SPI-7) where four primer pairs were used to detect tviA and tviB genes. Among 250 blood samples tested, 115 were positive by fliC PCR; 22 of these were negative for tviA and tviB. Hence, the method described here can be used to diagnose the incidence of Vi-negative serovar typhi especially in endemic regions where the Vi vaccine is administered. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Antimicrobial susceptibility pattern in children with typhoid fever and serotype of Salmonella typhi in Jakarta

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    Mirari Prasadajudio, Mulya Rahma Karyanti, Lia Waslia

    2017-03-01

    Full Text Available Objectives: Indonesia is known for high incidence of typhoid fever especially in children. This study aimed to observe antibiotic susceptibility in circulating Salmonella typhi serotypes in children with typhoid. Methods: A cross sectional study design was conducted. A total of 142 blood samples from children between 1-18 years old clinically diagnosed with suspected typhoid fever were recruited between January 2012 and July 2013 from six health centers in Jakarta. Confirmed cases were retrieved based on S. typhi isolate finding in blood culture. Antimicrobial susceptibility was investigated and PCR was used to detect S. typhi serotypes using fliB, fliC and aroC genes. Results: The prevalence of confirmed typhoid case based on isolate finding was 22 (15.5%. Twenty of S. typhi isolates expressed fliC gene carrying H:d allele, the other two expressed j allele, while only two samples expressed fliB, all showed no difference in pathogenicity and antimicrobial resistance. Conclusions: Circulating serotypes found in typhoid children in Jakarta, Indonesia are still susceptible even to the firstline antimicrobials. Thus, chloramphenicol, ampicillin and co-trimoxazole are still recommended. J Microbiol Infect Dis 2017; 7(1: 29-35

  16. A strand-specific RNA-Seq analysis of the transcriptome of the typhoid bacillus Salmonella typhi.

    Directory of Open Access Journals (Sweden)

    Timothy T Perkins

    2009-07-01

    Full Text Available High-density, strand-specific cDNA sequencing (ssRNA-seq was used to analyze the transcriptome of Salmonella enterica serovar Typhi (S. Typhi. By mapping sequence data to the entire S. Typhi genome, we analyzed the transcriptome in a strand-specific manner and further defined transcribed regions encoded within prophages, pseudogenes, previously un-annotated, and 3'- or 5'-untranslated regions (UTR. An additional 40 novel candidate non-coding RNAs were identified beyond those previously annotated. Proteomic analysis was combined with transcriptome data to confirm and refine the annotation of a number of hpothetical genes. ssRNA-seq was also combined with microarray and proteome analysis to further define the S. Typhi OmpR regulon and identify novel OmpR regulated transcripts. Thus, ssRNA-seq provides a novel and powerful approach to the characterization of the bacterial transcriptome.

  17. The Salmonella enterica serovar Typhi ltrR-ompR-ompC-ompF genes are involved in resistance to the bile salt sodium deoxycholate and in bacterial transformation.

    Science.gov (United States)

    Villarreal, J M; Becerra-Lobato, N; Rebollar-Flores, J E; Medina-Aparicio, L; Carbajal-Gómez, E; Zavala-García, M L; Vázquez, A; Gutiérrez-Ríos, R M; Olvera, L; Encarnación, S; Martínez-Batallar, A G; Calva, E; Hernández-Lucas, I

    2014-06-01

    A characterization of the LtrR regulator, an S. Typhi protein belonging to the LysR family is presented. Proteomics, outer membrane protein profiles and transcriptional analyses demonstrated that LtrR is required for the synthesis of OmpR, OmpC and OmpF. DNA-protein interaction analysis showed that LtrR binds to the regulatory region of ompR and then OmpR interacts with the ompC and ompF promoters inducing porin synthesis. LtrR-dependent and independent ompR promoters were identified, and both promoters are involved in the synthesis of OmpR for OmpC and OmpF production. To define the functional role of the ltrR-ompR-ompC-ompF genetic network, mutants in each gene were obtained. We found that ltrR, ompR, ompC and ompF were involved in the control of bacterial transformation, while the two regulators and ompC are necessary for the optimal growth of S. Typhi in the presence of one of the major bile salts found in the gut, sodium deoxycholate. The data presented establish the pivotal role of LtrR in the regulatory network of porin synthesis and reveal new genetic strategies of survival and cellular adaptation to the environment used by Salmonella. © 2014 John Wiley & Sons Ltd.

  18. Prevalence and susceptibility of salmonella Typhi and salmonella ...

    African Journals Online (AJOL)

    Methods: Blood samples collected from presumptive typhoid fever patients from Ahmadu Bello University (ABU), Federal College of Education (FCE) and presumptive typhoid fever patients that attended two private clinics (Salama Clinics and Savanna Polyclinics) in Zaria were cultured for Salmonella species and identified ...

  19. 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...... plasmid of Yersinia pestis....

  20. An evaluation of purified Salmonella Typhi protein antigens for the serological diagnosis of acute typhoid fever

    NARCIS (Netherlands)

    Tran Vu Thieu, Nga; Trinh van, Tan; Tran Tuan, Anh; Klemm, Elizabeth J.; Nguyen Ngoc Minh, Chau; Voong Vinh, Phat; Pham Thanh, Duy; Ho Ngoc Dan, Thanh; Pham Duc, Trung; Langat, Pinky; Martin, Laura B.; Galan, Jorge; Liang, Li; Felgner, Philip L.; Davies, D. Huw; de Jong, Hanna K.; Maude, Rapeephan R.; Fukushima, Masako; Wijedoru, Lalith; Ghose, Aniruddha; Samad, Rasheda; Dondorp, Arjen M.; Faiz, Abul; Darton, Thomas C.; Pollard, Andrew J.; Thwaites, Guy E.; Dougan, Gordon; Parry, Christopher M.; Baker, Stephen

    2017-01-01

    The diagnosis of typhoid fever is a challenge. Aiming to develop a typhoid diagnostic we measured antibody responses against Salmonella Typhi (S. Typhi) protein antigens and the Vi polysaccharide in a cohort of Bangladeshi febrile patients. IgM against 12 purified antigens and the Vi polysaccharide

  1. a rare case of salmonella typhi meningitis in an eleven month old ...

    African Journals Online (AJOL)

    Salmonella typhi is a rare cause of purulent meningitis. Since the earliest reviews on true typhoid meningitis by Cole in 1904 and Bayne Jones in 1917,sporadic case reports, mainly involving children have appeared in the literature. Only 9 cases of S. typhi meningitis in adults have been documented in the English literature.

  2. A Case Report of Acute Acalculous Cholecystitis and Acute Hemorrhagic Cystitis due to Salmonella Typhi

    Directory of Open Access Journals (Sweden)

    Hatice Beyazal Polat

    2014-01-01

    Full Text Available Acute acalculous cholecystitis and acute hemorrhagic cystitis due to Salmonella Typhi are a rare condition. A 24-year-old female patient was admitted to our clinic with abdominal pain, nausea, fever, headache, urinary burning, and bloody urine. Based on clinical, laboratory, and radiological evaluations, the patient was diagnosed with acute acalculous cholecystitis and acute hemorrhagic cystitis due to Salmonella Typhi. The patient was treated with intravenous ceftriaxone for two weeks. After the treatment, the patient's clinical and laboratory findings improved. Acute acalculous cholecystitis due to Salmonella Typhi concomitant with acute hemorrhagic cystitis is very rare and might be difficult to diagnose. Infectious agents such as Salmonella Typhi should be considered when acute acalculous cholecystitis and acute hemorrhagic cystitis are detected in adult patients with no underlying diseases.

  3. Variable Responses to Carbon Utilization between Planktonic and Biofilm Cells of a Human Carrier Strain of Salmonella enterica Serovar Typhi.

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    Kalaivani Kalai Chelvam

    Full Text Available Salmonella enterica serovar Typhi (S. Typhi is a foodborne pathogen that causes typhoid fever and infects only humans. The ability of S. Typhi to survive outside the human host remains unclear, particularly in human carrier strains. In this study, we have investigated the catabolic activity of a human carrier S. Typhi strain in both planktonic and biofilm cells using the high-throughput Biolog Phenotype MicroArray, Minimum Biofilm Eradication Concentration (MBEC biofilm inoculator (96-well peg lid and whole genome sequence data. Additional strains of S. Typhi were tested to further validate the variation of catabolism in selected carbon substrates in the different bacterial growth phases. The analyzes of the carbon utilization data indicated that planktonic cells of the carrier strain, S. Typhi CR0044 could utilize a broader range of carbon substrates compared to biofilm cells. Pyruvic acid and succinic acid which are related to energy metabolism were actively catabolised in the planktonic stage compared to biofilm stage. On the other hand, glycerol, L-fucose, L-rhamnose (carbohydrates and D-threonine (amino acid were more actively catabolised by biofilm cells compared to planktonic cells. Notably, dextrin and pectin could induce strong biofilm formation in the human carrier strain of S. Typhi. However, pectin could not induce formation of biofilm in the other S. Typhi strains. Phenome data showed the utilization of certain carbon substrates which was supported by the presence of the catabolism-associated genes in S. Typhi CR0044. In conclusion, the findings showed the differential carbon utilization between planktonic and biofilm cells of a S. Typhi human carrier strain. The differences found in the carbon utilization profiles suggested that S. Typhi uses substrates mainly found in the human biliary mucus glycoprotein, gallbladder, liver and cortex of the kidney of the human host. The observed diversity in the carbon catabolism profiles among

  4. A fast and highly sensitive blood culture PCR method for clinical detection of Salmonella enterica serovar Typhi

    Directory of Open Access Journals (Sweden)

    Zhou Liqing

    2010-04-01

    Full Text Available Abstract Background Salmonella Typhi causes an estimated 21 million new cases of typhoid fever and 216,000 deaths every year. Blood culture is currently the gold standard for diagnosis of typhoid fever, but it is time-consuming and takes several days for isolation and identification of causative organisms. It is then too late to initiate proper antibiotic therapy. Serological tests have very low sensitivity and specificity, and no practical value in endemic areas. As early diagnosis of the disease and prompt treatment are essential for optimal management, especially in children, a rapid sensitive detection method for typhoid fever is urgently needed. Although PCR is sensitive and rapid, initial research indicated similar sensitivity to blood culture and lower specificity. We developed a fast and highly sensitive blood culture PCR method for detection of Salmonella Typhi, allowing same-day initiation of treatment after accurate diagnosis of typhoid. Methods An ox bile tryptone soy broth was optimized for blood culture, which allows the complete lysis of blood cells to release intracellular bacteria without inhibiting the growth of Salmonella Typhi. Using the optimised broth Salmonella Typhi bacteria in artificial blood samples were enriched in blood culture and then detected by a PCR targeting the fliC-d gene of Salmonella Typhi. Results Tests demonstrated that 2.4% ox bile in blood culture not only lyzes blood cells completely within 1.5 hours so that the intracellular bacteria could be released, but also has no inhibiting effect on the growth of Salmonella Typhi. Three hour enrichment of Salmonella Typhi in tryptone soya broth containing 2.4% ox bile could increase the bacterial number from 0.75 CFU per millilitre of blood which is similar to clinical typhoid samples to the level which regular PCR can detect. The whole blood culture PCR assay takes less than 8 hours to complete rather than several days for conventional blood culture

  5. A Case Report of Acute Acalculous Cholecystitis and Acute Hemorrhagic Cystitis due to Salmonella Typhi

    OpenAIRE

    Beyazal Polat, Hatice; Beyazal, Mehmet; Beyazal Çeliker, Fatma

    2014-01-01

    Acute acalculous cholecystitis and acute hemorrhagic cystitis due to Salmonella Typhi are a rare condition. A 24-year-old female patient was admitted to our clinic with abdominal pain, nausea, fever, headache, urinary burning, and bloody urine. Based on clinical, laboratory, and radiological evaluations, the patient was diagnosed with acute acalculous cholecystitis and acute hemorrhagic cystitis due to Salmonella Typhi. The patient was treated with intravenous ceftriaxone for two weeks. After...

  6. Radiosensitization of Escherichia coli and Salmonella typhi in presence of active compounds

    Energy Technology Data Exchange (ETDEWEB)

    Lacroix, M. E-mail: monique.lacroix@inrs-iaf.uquebec.ca; Chiasson, F.; Borsa, J.; Ouattara, B

    2004-10-01

    The radiosensitization of Escherichia coli and Salmonella typhi in ground beef was evaluated in the presence of 18 active compounds. Medium fat ground beef (23% fat) was inoculated with E. coli or S. typhi and each active compound was added separately at various concentrations. For E. coli, the most efficient compounds were trans-cinnamaldehyde, thymol and thyme. For S. typhi, the most efficient compounds was trans-cinnamaldehyde, carvacrol and thymol. The addition of tetrasodium pyrophosphate, carvacrol and ascorbic acid had no effect on the irradiation sensitivity of E. coli. For S. typhi, only ascorbic acid had no effect.

  7. Active protection of mice against Salmonella typhi by immunization with strain-specific porins.

    Science.gov (United States)

    Isibasi, A; Ortiz-Navarrete, V; Paniagua, J; Pelayo, R; González, C R; García, J A; Kumate, J

    1992-01-01

    NIH mice were immunized with between 2.5 and 30 micrograms of two highly purified porins, 34 kDa and 36 kDa, isolated from the virulent strain Salmonella typhi 9,12, Vi:d. Of mice immunized with 10 micrograms of porins, 90% were protected against a challenge with up to 500 LD50 (50% lethal doses) of S. typhi 9,12,Vi:d and only 30% protection was observed in mice immunized with the same dose of porins but challenged with the heterologous strain Salmonella typhimurium. These results demonstrate the utility of porins for the induction of a protective status against S. typhi in mice.

  8. Transcriptional regulation of the assT-dsbL-dsbI gene cluster in Salmonella enterica serovar Typhi IMSS-1 depends on LeuO, H-NS, and specific growth conditions.

    Science.gov (United States)

    Gallego-Hernández, A L; Hernández-Lucas, I; De la Cruz, M A; Olvera, L; Morett, E; Medina-Aparicio, L; Ramírez-Trujillo, J A; Vázquez, A; Fernández-Mora, M; Calva, E

    2012-05-01

    The assT gene encodes an arylsulfate sulfotransferase, an enzyme that catalyzes sulfuryl transfer from phenolic sulfate to a phenolic acceptor. In Salmonella enterica serovar Typhi IMSS-1, the assT gene is located upstream of the dsbL and dsbI genes, which are involved in a disulfide bond formation required for its activation. The assT-dsbL-dsbI gene cluster forms an operon transcribed by a LeuO-dependent promoter, in rich medium A (MA). Interestingly, in the absence of cloned leuO and in a ΔleuO background, two transcription start sites were detected for assT and two for dsbL-dsbI in minimal medium. The H-NS nucleoid protein repressed the expression of the assT-dsbL-dsbI LeuO-dependent operon, as well as of the assT transcriptional units. Thus, the expression of the assT-dsbL-dsbI gene cluster depends on the global regulatory proteins LeuO and H-NS, as well as on specific growth conditions.

  9. Induction of resistant mutants of Salmonella enterica serotype Typhi under ciprofloxacin selective pressure

    Directory of Open Access Journals (Sweden)

    Sushila Dahiya

    2014-01-01

    Full Text Available Background & objectives: Infection with Salmonella enterica serovar Typhi (hereafter S. Typhi is an important public health problem in India. There has been an increase in the number of reported clinical failures to ciprofloxacin treatment but the data on possible mechanism of failure are limited. One mechanism that has been widely reported and found associated with ciprofloxacin resistance, is the mutations in target genes in QRDR (quinolone resistance determining region. It is hypothesized that mutations in DNA gyrase or topoisomerase IV result in therapeutic failure under selective pressure of antibiotic while the patient is on treatment. We undertook in vitro sequential selection studies to expose the clinical isolates of S. Typhi to different concentration of ciprofloxacin to study the role of antibiotic selective pressure in the development of mutations in QRDR. Methods: Total 26 clinical isolates were divided in to two parts: part I included six isolates obtained from three patients with relapse of enteric fever and part II included 20 isolates with different ciprofloxacin MIC levels. For in vitro induction of mutation experiment, five S. Typhi isolates were selected which included three NAS (nalidixic acid sensitive and 2 NAR (nalidixic acid resistant S. Typhi. These isolates were grown under increasing concentrations of ciprofloxacin and mutations acquired in QRDR of DNA gyrase (gyrA and gyrB and topoisomerase IV (parC and parE were investigated by sequencing. Results: For the isolates included in the part I of the study, it was found that the MIC to ciprofloxacin increased in the isolates obtained during the relapse of enteric fever as compare to the first isolate. All isolates had single mutation in gyrA gene at S83 without additional mutation in the second isolate. In the second part of the study, the nine isolates with varying MICs to ciprofloxacin also had single mutation in gyrA gene at S83 and another six had triple mutations

  10. Nanoparticle Fullerene (C60) demonstrated stable binding with antibacterial potential towards probable targets of drug resistant Salmonella typhi - a computational perspective and in vitro investigation.

    Science.gov (United States)

    Skariyachan, Sinosh; Parveen, Asma; Garka, Shruti

    2017-12-01

    Salmonella typhi, a Gram negative bacterium, has become multidrug resistant (MDR) to wide classes of antibacterials which necessitate an alarming precaution. This study focuses on the binding potential and therapeutic insight of Nano-Fullerene C60 towards virulent targets of Salmonella typhi by computational prediction and preliminary in vitro assays. The clinical isolates of Salmonella typhi were collected and antibiotic susceptibility profiles were assessed. The drug targets of pathogen were selected by rigorous literature survey and gene network analysis by various metabolic network resources. Based on this study, 20 targets were screened and the 3D structures of few drug targets were retrieved from PDB and others were computationally predicted. The structures of nanoleads such as Fullerene C60, ZnO and CuO were retrieved from drug databases. The binding potential of these nanoleads towards all selected targets were predicted by molecular docking. The best docked conformations were screened and concept was investigated by preliminary bioassays. This study revealed that most of the isolates of Salmonella typhi were found to be MDR (p C60 showed better binding affinity towards the drug targets when compared to ZnO and CuO. The preliminary in vitro assays suggested that 100 μg/L Fullerene C60 posses significant inhibitory activities and absence of drug resistance to this nanoparticle. This study suggests that Fullerene C60 can be scaled up as probable lead molecules against the major drug targets of MDR Salmonella typhi.

  11. Protection against Salmonella typhi infection in mice after immunization with outer membrane proteins isolated from Salmonella typhi 9,12,d, Vi.

    Science.gov (United States)

    Isibasi, A; Ortiz, V; Vargas, M; Paniagua, J; González, C; Moreno, J; Kumate, J

    1988-11-01

    The current studies were undertaken to assess the ability of the outer membrane proteins (OMPs) of Salmonella typhi to induce protection against challenge with the bacteria in mucin. OMPs were isolated as described by Schnaitman (J. Bacteriol. 108:553-556, 1971) and were found to be contaminated with approximately 4% lipopolysaccharide (LPS). Immunization with as little as 30 micrograms of OMPs conferred 100% protection to mice challenged with up to 1,000 50% lethal doses (LD50) of two strains of S. typhi (9,12,d, Vi and Ty2). In addition, 30% protection against challenge with up to 500 LD50 of Salmonella typhimurium was achieved. Immunization with LPS at doses equivalent to those found in the OMPs was considerably inferior to the OMPs in the induction of an immune status. Moreover, LPS was effective only when the challenge was performed with S. typhi 9,12,d, Vi (40% protection to 100 LD50). An antiserum raised in rabbits reacted mainly against the bands of the molecular weights corresponding to the so-called porins contained in the OMP preparation as shown by Western blotting (immunoblotting). This rabbit antiserum protected 100% of mice against challenge with 100 LD50 of either strain of S. typhi and 80% of mice against challenge with the same LD50 of S. typhimurium. These results indicate the usefulness of OMPs in the induction of active immunity against S. typhi in mice.

  12. Ekstrak Pegagan Meningkatkan Titer Antibodi Mencit Setelah Diinfeksi Salmonella typhi (CENTELLA ASIATICA EXTRACT INCREASE ANTIBODY TITER IN MICE AFTER SALMONELLA TYPHI INFECTION

    Directory of Open Access Journals (Sweden)

    I Nengah Kerta Besung

    2013-09-01

    Full Text Available A study was conducted to find out the ability of Centella asiatica (C. asiatica in enhancing antibodyresponse of C. asiatica treated mice following Salmonella typhi (S. typhi infections. It is therefore expectedthat herbal drug such as  C. asiatica  can be used as an alternative medicine to prevent and to curesalmonellosis both in animals and human. Experimental laboratory studies were conducted usingCompletely Factorial Randomized Design. Mice were divided into four groups and they were treatedrespectively with destilated water (negative control, 125, 250, and 500 mg/kg BW/day of  C. asiaticaextract. The treatment was conducted daily for two weeks  and the mice were inoculated with 105 cells/mlof  S. typhi. The antibody response were examined by indirect enzyme-linked immunosorbent assay (ELISAon first day, second week and fourth week  after S. typhi infections.  The result showed that treatment ofmice with C. asiatica extract significantly (p<0,05 enhanced antibody titer of Balb/c mice after S. typhiinfections. The highest antibody titer was observed at four weeks after S. typhi infections with 500 mg/kgBW/day (94,0370 ± 1,69 IU.

  13. Modification Of Carry-Blair Transport Media For Storage Salmonella typhi

    Directory of Open Access Journals (Sweden)

    Yati Supriatin

    2016-09-01

    Full Text Available The aim of this study was to determine transport media modification as alternative media to replace Carry Blair. One type of transport media that often use to carry faeces specimens suspected to contain Salmonella typhi is Carry-Blair media. Studies have been conducted experimentally by storing Salmonella typhi on alternative transport media with Peptone composition, disodium Phosphate, Sodium chloride, Calcium chloride, which is made using a semi-solid and Carry-Blair as a control. Three variety of storage was done (0 hour,6 hours,9 hours at a temperature 4⁰-8⁰C and then Salmonella typhi was inoculated in Salmonella Shigella Agar using spread plate technique incubated during 24 hours at 37⁰C, counted the number of colonies by the plate count method using the colony counter. The results of ANOVA could be concluded that modification media could be use as alternative media replace Carry-Blair at 6 hours. Based on regression correlation test was assumed that the Salmonella typhi bacteria still life at less than 11 hours 54 minutes.

  14. blaCTX-M-I group extended spectrum beta lactamase-producing Salmonella typhi from hospitalized patients in Lagos, Nigeria

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    Akinyemi KO

    2015-05-01

    Full Text Available Kabiru O Akinyemi,1 Bamidele A Iwalokun,2 Olajide O Alafe,1 Sulaiman A Mudashiru,1 Christopher Fakorede,11Department of Microbiology, Lagos State University, Ojo, Lagos, Nigeria; 2Biochemistry and Nutrition Division, Nigerian Institute of Medical Research, Yaba, Lagos, NigeriaPurpose: The global spread of blaCTX-M-I extended-spectrum beta-lactamase (ESBL-producing Salmonella spp. remains a major threat to treatment and control. Evidence of emergence and spread of this marker are lacking in Nigeria. This study investigated blaCTX-M-I ESBL production among Salmonella isolates from hospitalized patients.Methods: Patients (158 total made up of two groups were evaluated. Group A was composed of 135 patients with persistent pyrexia and group B was composed of 23 gastroenteritis patients and their stool samples. Samples were cultured, and isolates were identified and were subjected to antibiotic susceptibility testing by standard methods. Isolates were further screened for ESBL production, blaCTX-M-I genes and transferability by double disk synergy test, plasmid extraction, polymerase chain reaction, and conjugation experiment.Results: Thirty-five (25.9% Salmonella isolates were identified from group A, of which 74.3% were S. typhi, 22.9% were S. paratyphi and two (5.7% were invasive non-typhoidal S. enteritidis. Nine Plasmodium falciparum infections were recorded, four of which were identified as co-infections with typhoidal Salmonella. Only two (8.7% S. enteritidis samples were obtained from group B (P>0.05. A total of 24 isolates were ESBL-positive, eliciting resistance to five to seven antibiotics, and were multiple-drug resistant. ESBL production due to the blaCTX-M-I gene cluster was detected in eleven (45.8% Salmonella isolates. Nine (81.8% of the eleven blaCTX-M-I ESBL producers were S. typhi and two (18.2% isolates were S. enteritidis. Four of nine S. typhi blaCTX-M-I ESBL-producing strains harbored 23 kb self-transmissible plasmid that was co

  15. Protection against Salmonella typhi infection in mice after immunization with outer membrane proteins isolated from Salmonella typhi 9,12,d, Vi.

    OpenAIRE

    Isibasi, A.; ORTIZ,V; Vargas, M.; Paniagua, J; GONZÁLEZ, C.1,3; Moreno, J.; Kumate, J.

    1988-01-01

    The current studies were undertaken to assess the ability of the outer membrane proteins (OMPs) of Salmonella typhi to induce protection against challenge with the bacteria in mucin. OMPs were isolated as described by Schnaitman (J. Bacteriol. 108:553-556, 1971) and were found to be contaminated with approximately 4% lipopolysaccharide (LPS). Immunization with as little as 30 micrograms of OMPs conferred 100% protection to mice challenged with up to 1,000 50% lethal doses (LD50) of two strain...

  16. A novel method to generate Salmonella Typhi Ty21a ghosts exploiting the λ phage holin-endolysin system.

    Science.gov (United States)

    Won, Gayeon; Kim, Boram; Lee, John Hwa

    2017-07-18

    Human typhoid fever caused by Salmonella Typhi still poses a severe global disease burden in developing countries despite the availability of commercial vaccines. In this study, we constructed a non-living S. Typhi Ty21a vaccine candidate by employing a lambda (λ) phage-derived holin-endolysin system to efficiently construct bacterial ghosts. The lysis plasmid pJHL464 harbors an R lysis cassette that is stringently regulated by dual promoters containing cI857/λPR and ParaBAD/araC components. The plasmid was introduced into an asd gene-deleted S. Typhi Ty21a strain designated JOL1675. The in vitro expression of endolysin (~17.76 kDa) in the subsequent JOL1675 vaccine construct when grown under lysis inducible conditions was validated by immunoblotting. In scanning electron microscopy analysis, surface transmembrane tunnels and a collapsed body were visualized in the ghosts. Following 48 h of lysis, no viable JOL1675 cells remained, indicating that lysis of all cells was achieved. Subcutaneous immunizations of mice with the JOL1675 ghosts produced significantly increasing titers of serum IgG and vaginal wash secretory IgA antibodies against JOL1675 outer membrane proteins during the observational period. Further, serum collected at 6 weeks post-immunization of rabbits exhibited effective bactericidal activity against wild type S. Typhi in the presence of complement. These data showed that JOL1675 ghosts are highly immunogenic and elicit humoral and mucosal responses expected to correlate with protective immunity against S. typhi. Collectively, our findings support the conclusion that incorporating a λ phage holin-endolysin-mediated lysis construct into S. Typhi is an efficient strategy for developing a novel and safe non-living typhoid vaccine candidate.

  17. IMPACT OF FOOD AND FOLATE SUPPLEMENTATION DURING Salmonella TYPHI INFECTION IN Caenorhabditis elegans

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    Bhagavathi Sundaram Sivamaruthi

    2012-06-01

    Full Text Available Caenorhabditis elegans is an instructive and suitable model for studying pathogenesis of almost all human pathogens. Salmonella Typhi is gram-negative facultative intracellular anaerobe that causes several pathetic infections. Necessary enriched nutrient ingestion during pathological conditions may reduce the harshness of the infection. We investigated the impact of folate and food supplementation during S. Typhi infection on the model system, C. elegans. Our data indicated that folate supplementation (10 µg increases the lifespan of S. Typhi infected C. elegans up to 20%. In combination with laboratory food source E. coli OP50, folate increases the infected the worm’s lifespan to 40%. The wild type C. elegans infected by S. Typhi died with the LT50 of 60 ± 12 h. The LT50 of S. Typhi infected folt-1 mutant strain VC959 was 96 ± 6 h. However, the folate supplemented mutant worms exhibited an extended life with LT50 of 120 ± 6 h. The short time exposure and pharyngeal pumping studies confirmed that folt-1 mutant worm exhibited increased survival rate during pathogenic course at significant level when compared to wild-type. Our data revealed that folt-1 plays a significant role in host defense system against S. Typhi infection and the folate supplementation in combination with food increases the host survival during S. Typhi infection.

  18. Salmonella Typhi sense host neuroendocrine stress hormones and release the toxin haemolysin E

    Science.gov (United States)

    Karavolos, Michail H; Bulmer, David M; Spencer, Hannah; Rampioni, Giordano; Schmalen, Ira; Baker, Stephen; Pickard, Derek; Gray, Joe; Fookes, Maria; Winzer, Klaus; Ivens, Alasdair; Dougan, Gordon; Williams, Paul; Khan, C M Anjam

    2011-01-01

    Salmonella enterica serovar Typhi (S. typhi) causes typhoid fever. We show that exposure of S. typhi to neuroendocrine stress hormones results in haemolysis, which is associated with the release of haemolysin E in membrane vesicles. This effect is attributed to increased expression of the small RNA micA and RNA chaperone Hfq, with concomitant downregulation of outer membrane protein A. Deletion of micA or the two-component signal-transduction system, CpxAR, abolishes the phenotype. The hormone response is inhibited by the β-blocker propranolol. We provide mechanistic insights into the basis of neuroendocrine hormone-mediated haemolysis by S. typhi, increasing our understanding of inter-kingdom signalling. PMID:21331094

  19. OPTIMATION OF 48 KHZ ULTRASONIC WAVE DOSE FOR THE INACTIVATION OF SALMONELLA TYPHI

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    Dwi May Lestari

    2015-01-01

    Full Text Available This study was aimed to determine the effect of ultrasonic dose exposure which could decrease the viability of Salmonella typhi by using the variation of exposure time (15, 20, 25, and 30 minutes and volume of bacterial suspension (2, 4, 6, and 8 ml at constant power. The sample used was Salmonella typhi. Ultrasonic wave transmitter was a piezoelectric tweeter with 0,191 watts of power and 48 kHz frequency generated by the signal generator. Piezoelectric tweeter was a kind of transducer which converted electrical energy into ultrasonic energy. This research was an experimental laboratory with a completely randomized design. The decrease of bacterial percentage was calculated by using TPC (Total Plate Count. Data were analyzed by using One Way Anova. The results showed that the variation of exposure time and volume of bacterial suspension gave significant effect on the percentage of Salmonella typhi kill. The most optimal of ultrasonic dose exposure to kill Salmonella typhi was 281.87 J/ml with 100% bacterial kill.

  20. Detection of Salmonella typhi by nested polymerase chain reaction in blood, urine, and stool samples

    NARCIS (Netherlands)

    Hatta, Mochammad; Smits, Henk L.

    2007-01-01

    A nested polymerase chain reaction (PCR) specific for Salmonella enterica serovar Typhi was used for the detection of the pathogen in blood, urine, and stool samples from 131 patients with clinical suspicion of typhoid fever. The sensitivity of blood culture, the PCRs with blood, urine, and feces,

  1. Non-typhi salmonella in children with severe malaria | Oundo | East ...

    African Journals Online (AJOL)

    19,118 (7.3%) and 342/1,820 (19%) respectively. Non-typhi salmonella consisted of 260/1,395 (18.6%) of the positive blood cultures and 92/324 (28.4%) of the stool cultures out of which a total of 101 NTS occurred in children with severe ...

  2. Gold nanoparticles as efficient antimicrobial agents for Escherichia coli and Salmonella typhi

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    Lima Enrique

    2013-01-01

    Full Text Available Abstract Background It is imperative to eliminate bacteria present in water in order to avoid problems in healthy. Escherichia coli and Salmonella typhi bacteria are two common pollutants and they are developing resistance to some of the most used bactericide. Therefore new biocide materials are being tested. Thus, gold nanoparticles are proposed to inhibit the growth of these two microorganisms. Results Gold nanoparticles were supported onto clinoptilolite, mordenite and faujasite zeolites. Content of gold in materials varied between 2.3 and 2.8 wt%. The size, dispersion and roughness of gold nanoparticles were highly dependent of the zeolite support. The faujasite support was the support where the 5 nm nanoparticles were highly dispersed. The efficiency of gold-zeolites as bactericides of Escherichia coli and Salmonella typhi was determined by the zeolite support. Conclusions Gold nanoparticles dispersed on zeolites eliminate Escherichia coli and Salmonella typhi at short times. The biocidal properties of gold nanoparticles are influenced by the type of support which, indeed, drives key parameters as the size and roughness of nanoparticles. The more actives materials were pointed out Au-faujasite. These materials contained particles sized 5 nm at surface and eliminate 90–95% of Escherichia coli and Salmonella typhi colonies.

  3. Prevalence of Salmonella typhi and intestinal parasites among food ...

    African Journals Online (AJOL)

    Bernt Lindtjorn

    Abstract. Background: Food borne diseases are a global public health problem. Food handlers play a major role for the transmission of food borne diseases. Objectives: This study was aimed at exploring the prevalence of intestinal parasites, S. typhi carrier rate and risk factors among food handlers at Bahir Dar town.

  4. Prevalence of Salmonella typhi and intestinal parasites among food ...

    African Journals Online (AJOL)

    Background: Food borne diseases are a global public health problem. Food handlers play a major role for the transmission of food borne diseases. Objectives: This study was aimed at exploring the prevalence of intestinal parasites, S. typhi carrier rate and risk factors among food handlers at Bahir Dar town. Methods: A ...

  5. Clearance and tissue distribution of intravenously injected Salmonella typhi polysaccharide in rabbits.

    OpenAIRE

    Isibasi, A.; Jimenez, E.; Kumate, J.

    1983-01-01

    The interaction of Freeman polysaccharide of Salmonella typhi with blood and tissues of rabbits was studied by radioimmunoassay. After intravenous injection of 1.0 mg of S. typhi Freeman polysaccharide, a rapid clearance phase (t1/2, 6.0 min) was followed by a slower clearance period (t1/2, 55.2 min). These results suggest first, that the distribution of whole lipopolysaccharide is a function of how the polysaccharides are handled by the host; further, that the O side chain determines how and...

  6. Prevalence of current patterns and predictive trends of multidrug-resistant Salmonella Typhi in Sudan

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    Ayman A. Elshayeb

    2017-11-01

    Full Text Available Abstract Background Enteric fever has persistence of great impact in Sudanese public health especially during rainy season when the causative agent Salmonella enterica serovar Typhi possesses pan endemic patterns in most regions of Sudan - Khartoum. Objectives The present study aims to assess the recent state of antibiotics susceptibility of Salmonella Typhi with special concern to multidrug resistance strains and predict the emergence of new resistant patterns and outbreaks. Methods Salmonella Typhi strains were isolated and identified according to the guidelines of the International Standardization Organization and the World Health Organization. The antibiotics susceptibilities were tested using the recommendations of the Clinical Laboratories Standards Institute. Predictions of emerging resistant bacteria patterns and outbreaks in Sudan were done using logistic regression, forecasting linear equations and in silico simulations models. Results A total of 124 antibiotics resistant Salmonella Typhi strains categorized in 12 average groups were isolated, different patterns of resistance statistically calculated by (y = ax − b. Minimum bactericidal concentration’s predication of resistance was given the exponential trend (y = n ex and the predictive coefficient R2 > 0 < 1 are approximately alike. It was assumed that resistant bacteria occurred with a constant rate of antibiotic doses during the whole experimental period. Thus, the number of sensitive bacteria decreases at the same rate as resistant occur following term to the modified predictive model which solved computationally. Conclusion This study assesses the prediction of multi-drug resistance among S. Typhi isolates by applying low cost materials and simple statistical methods suitable for the most frequently used antibiotics as typhoid empirical therapy. Therefore, bacterial surveillance systems should be implemented to present data on the aetiology and current

  7. A 23-year retrospective investigation of Salmonella Typhi and Salmonella Paratyphi isolated in a tertiary Kathmandu hospital.

    Science.gov (United States)

    Zellweger, Raphaël M; Basnyat, Buddha; Shrestha, Poojan; Prajapati, Krishna G; Dongol, Sabina; Sharma, Paban K; Koirala, Samir; Darton, Thomas C; Dolecek, Christiane; Thompson, Corinne N; Thwaites, Guy E; Baker, Stephen G; Karkey, Abhilasha

    2017-11-01

    Salmonella serovars Typhi (S. Typhi) and Paratyphi A (S. Paratyphi A), the causative agents of enteric fever, have been routinely isolated organisms from the blood of febrile patients in the Kathmandu Valley since the early 1990s. Susceptibility against commonly used antimicrobials for treating enteric fever has gradually changed throughout South Asia since this time, posing serious treatment challenges. Here, we aimed to longitudinally describe trends in the isolation of Salmonella enterica and assess changes in their antimicrobial susceptibility in Kathmandu over a 23-year period. We conducted a retrospective analysis of standardised microbiological data from April 1992 to December 2014 at a single healthcare facility in Kathmandu, examining time trends of Salmonella-associated bacteraemia and the corresponding antimicrobial susceptibility profiles of the isolated organisms. Over 23 years there were 30,353 positive blood cultures. Salmonella enterica accounted for 65.4% (19,857/30,353) of all the bacteria positive blood cultures. S. Typhi and S. Paratyphi A were the dominant serovars, constituting 68.5% (13,592/19,857) and 30.5% (6,057/19,857) of all isolated Salmonellae. We observed (i) a peak in the number of Salmonella-positive cultures in 2002, a year of heavy rainfall and flooding in the Kathmandu Valley, followed by a decline toward pre-flood baseline by 2014, (ii) an increase in the proportion of S. Paratyphi in all Salmonella-positive cultures between 1992 and 2014, (iii) a decrease in the prevalence of MDR for both S. Typhi and S. Paratyphi, and (iv) a recent increase in fluoroquinolone non-susceptibility in both S. Typhi and S. Paratyphi isolates. Our work describes significant changes in the epidemiology of Salmonella enterica in the Kathmandu Valley during the last quarter of a century. We highlight the need to examine current treatment protocols for enteric fever and suggest a change from fluoroquinolone monotherapy to combination therapies of

  8. A 23-year retrospective investigation of Salmonella Typhi and Salmonella Paratyphi isolated in a tertiary Kathmandu hospital.

    Directory of Open Access Journals (Sweden)

    Raphaël M Zellweger

    2017-11-01

    Full Text Available Salmonella serovars Typhi (S. Typhi and Paratyphi A (S. Paratyphi A, the causative agents of enteric fever, have been routinely isolated organisms from the blood of febrile patients in the Kathmandu Valley since the early 1990s. Susceptibility against commonly used antimicrobials for treating enteric fever has gradually changed throughout South Asia since this time, posing serious treatment challenges. Here, we aimed to longitudinally describe trends in the isolation of Salmonella enterica and assess changes in their antimicrobial susceptibility in Kathmandu over a 23-year period.We conducted a retrospective analysis of standardised microbiological data from April 1992 to December 2014 at a single healthcare facility in Kathmandu, examining time trends of Salmonella-associated bacteraemia and the corresponding antimicrobial susceptibility profiles of the isolated organisms.Over 23 years there were 30,353 positive blood cultures. Salmonella enterica accounted for 65.4% (19,857/30,353 of all the bacteria positive blood cultures. S. Typhi and S. Paratyphi A were the dominant serovars, constituting 68.5% (13,592/19,857 and 30.5% (6,057/19,857 of all isolated Salmonellae. We observed (i a peak in the number of Salmonella-positive cultures in 2002, a year of heavy rainfall and flooding in the Kathmandu Valley, followed by a decline toward pre-flood baseline by 2014, (ii an increase in the proportion of S. Paratyphi in all Salmonella-positive cultures between 1992 and 2014, (iii a decrease in the prevalence of MDR for both S. Typhi and S. Paratyphi, and (iv a recent increase in fluoroquinolone non-susceptibility in both S. Typhi and S. Paratyphi isolates.Our work describes significant changes in the epidemiology of Salmonella enterica in the Kathmandu Valley during the last quarter of a century. We highlight the need to examine current treatment protocols for enteric fever and suggest a change from fluoroquinolone monotherapy to combination therapies

  9. OmpR may regulate the putative YehU/YehT two-component system in Salmonella enterica serovar Typhi under hypotonic growth condition.

    Science.gov (United States)

    Zhang, Haifang; Du, Hong; Ji, Xiaolan; Ni, Bin; Mao, Linxiang; Xu, Shungao; Sheng, Xiumei; Xu, Huaxi; Huang, Xinxiang

    2012-03-01

    Decreased expression (twofold) of a putative yehUTS operon of which yehUT encodes a putative YehU/YehT two-component system in the ompR mutant from Salmonella enterica serovar Typhi (S. Typhi) GIFU10007 under hypotonic growth condition was observed by qRT-PCR. Purified recombinant protein OmpR(His6) of GIFU10007 was shown to bind the upstream region of the yehU gene by the gel-shift assay. In addition, the yehT deletion mutant (ΔyehT) displayed differential expression (twofold or higher) of 26 genes under the condition by the DNA microarray analysis. Altogether, OmpR might regulate the YehUT system in S. Typhi under hypotonic growth condition.

  10. [Multifocal osteoarticular infection caused by Salmonella non typhi in a child with sickle cell disease].

    Science.gov (United States)

    Cabaret, B; Couëc, M-L; Lorrot, M; Launay, E; Gras-Le Guen, C

    2013-04-01

    Sickle cell disease is the most common monogenic hereditary hemoglobinopathy. Its course is marked by vaso-occlusive crises (VOC), episodes of acute hemolytic anemia on a background of chronic hemolytic anemia, and severe infections. A 2-year-old child with sickle cell disease presented with severe sepsis caused by Salmonella non typhi. Control of the sepsis was difficult, with multifocal osteomyelitis and arthritis, which required prolonged intravenous antibiotic therapy. Prolonged treatment was complicated by cardiorespiratory arrest and severe neurological damage, as well as nosocomial infections. Osseous articular infections caused by Salmonella non typhi are a common complication in children with sickle cell disease, which need to be promptly recognized. Management remains a great concern. The clinical case reported herein is original in its multifocal evolution. It illustrates the vulnerability of patients with sickle cell disease and the need for urgent and intensive care in the case of infection. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  11. Role of Environmental Factors in Shaping Spatial Distribution of Salmonella enterica Serovar Typhi, Fiji.

    Science.gov (United States)

    de Alwis, Ruklanthi; Watson, Conall; Nikolay, Birgit; Lowry, John H; Thieu, Nga Tran Vu; Van, Tan Trinh; Ngoc, Dung Tran Thi; Rawalai, Kitione; Taufa, Mere; Coriakula, Jerimaia; Lau, Colleen L; Nilles, Eric J; Edmunds, W John; Kama, Mike; Baker, Stephen; Cano, Jorge

    2018-02-01

    Fiji recently experienced a sharp increase in reported typhoid fever cases. To investigate geographic distribution and environmental risk factors associated with Salmonella enterica serovar Typhi infection, we conducted a cross-sectional cluster survey with associated serologic testing for Vi capsular antigen-specific antibodies (a marker for exposure to Salmonella Typhi in Fiji in 2013. Hotspots with high seroprevalence of Vi-specific antibodies were identified in northeastern mainland Fiji. Risk for Vi seropositivity increased with increased annual rainfall (odds ratio [OR] 1.26/quintile increase, 95% CI 1.12-1.42), and decreased with increased distance from major rivers and creeks (OR 0.89/km increase, 95% CI 0.80-0.99) and distance to modeled flood-risk areas (OR 0.80/quintile increase, 95% CI 0.69-0.92) after being adjusted for age, typhoid fever vaccination, and home toilet type. Risk for exposure to Salmonella Typhi and its spatial distribution in Fiji are driven by environmental factors. Our findings can directly affect typhoid fever control efforts in Fiji.

  12. Cell-free culture supernatant of Bifidobacterium breve CNCM I-4035 decreases pro-inflammatory cytokines in human dendritic cells challenged with Salmonella typhi through TLR activation.

    Directory of Open Access Journals (Sweden)

    Miriam Bermudez-Brito

    Full Text Available Dendritic cells (DCs constitute the first point of contact between gut commensals and our immune system. Despite growing evidence of the immunomodulatory effects of probiotics, the interactions between the cells of the intestinal immune system and bacteria remain largely unknown. Indeed,, the aim of this work was to determine whether the probiotic Bifidobacterium breve CNCM I-4035 and its cell-free culture supernatant (CFS have immunomodulatory effects in human intestinal-like dendritic cells (DCs and how they respond to the pathogenic bacterium Salmonella enterica serovar Typhi, and also to elucidate the molecular mechanisms involved in these interactions. Human DCs were directly challenged with B. breve/CFS, S. typhi or a combination of these stimuli for 4 h. The expression pattern of genes involved in Toll-like receptor (TLR signaling pathway and cytokine secretion was analyzed. CFS decreased pro-inflammatory cytokines and chemokines in human intestinal DCs challenged with S. typhi. In contrast, the B. breve CNCM I-4035 probiotic strain was a potent inducer of the pro-inflammatory cytokines and chemokines tested, i.e., TNF-α, IL-8 and RANTES, as well as anti-inflammatory cytokines including IL-10. CFS restored TGF-β levels in the presence of Salmonella. Live B.breve and its supernatant enhanced innate immune responses by the activation of TLR signaling pathway. These treatments upregulated TLR9 gene transcription. In addition, CFS was a more potent inducer of TLR9 expression than the probiotic bacteria in the presence of S. typhi. Expression levels of CASP8 and IRAK4 were also increased by CFS, and both treatments induced TOLLIP gene expression. Our results indicate that the probiotic strain B. breve CNCM I-4035 affects the intestinal immune response, whereas its supernatant exerts anti-inflammatory effects mediated by DCs. This supernatant may protect immune system from highly infectious agents such as Salmonella typhi and can down

  13. Cell-Free Culture Supernatant of Bifidobacterium breve CNCM I-4035 Decreases Pro-Inflammatory Cytokines in Human Dendritic Cells Challenged with Salmonella typhi through TLR Activation

    Science.gov (United States)

    Bermudez-Brito, Miriam; Muñoz-Quezada, Sergio; Gomez-Llorente, Carolina; Matencio, Esther; Bernal, Maria J.; Romero, Fernando; Gil, Angel

    2013-01-01

    Dendritic cells (DCs) constitute the first point of contact between gut commensals and our immune system. Despite growing evidence of the immunomodulatory effects of probiotics, the interactions between the cells of the intestinal immune system and bacteria remain largely unknown. Indeed,, the aim of this work was to determine whether the probiotic Bifidobacterium breve CNCM I-4035 and its cell-free culture supernatant (CFS) have immunomodulatory effects in human intestinal-like dendritic cells (DCs) and how they respond to the pathogenic bacterium Salmonella enterica serovar Typhi, and also to elucidate the molecular mechanisms involved in these interactions. Human DCs were directly challenged with B. breve/CFS, S. typhi or a combination of these stimuli for 4 h. The expression pattern of genes involved in Toll-like receptor (TLR) signaling pathway and cytokine secretion was analyzed. CFS decreased pro-inflammatory cytokines and chemokines in human intestinal DCs challenged with S. typhi. In contrast, the B. breve CNCM I-4035 probiotic strain was a potent inducer of the pro-inflammatory cytokines and chemokines tested, i.e., TNF-α, IL-8 and RANTES, as well as anti-inflammatory cytokines including IL-10. CFS restored TGF-β levels in the presence of Salmonella. Live B.breve and its supernatant enhanced innate immune responses by the activation of TLR signaling pathway. These treatments upregulated TLR9 gene transcription. In addition, CFS was a more potent inducer of TLR9 expression than the probiotic bacteria in the presence of S. typhi. Expression levels of CASP8 and IRAK4 were also increased by CFS, and both treatments induced TOLLIP gene expression. Our results indicate that the probiotic strain B. breve CNCM I-4035 affects the intestinal immune response, whereas its supernatant exerts anti-inflammatory effects mediated by DCs. This supernatant may protect immune system from highly infectious agents such as Salmonella typhi and can down-regulate pro

  14. Clinically and Microbiologically Derived Azithromycin Susceptibility Breakpoints for Salmonella enterica Serovars Typhi and Paratyphi A

    Science.gov (United States)

    Thieu, Nga Tran Vu; Dolecek, Christiane; Karkey, Abhilasha; Gupta, Ruchi; Turner, Paul; Dance, David; Maude, Rapeephan R.; Ha, Vinh; Tran, Chinh Nguyen; Thi, Phuong Le; Be, Bay Pham Van; Phi, La Tran Thi; Ngoc, Rang Nguyen; Ghose, Aniruddha; Dongol, Sabina; Campbell, James I.; Thanh, Duy Pham; Thanh, Tuyen Ha; Moore, Catrin E.; Sona, Soeng; Gaind, Rajni; Deb, Monorama; Anh, Ho Van; Van, Sach Nguyen; Tinh, Hien Tran; Day, Nicholas P. J.; Dondorp, Arjen; Thwaites, Guy; Faiz, Mohamed Abul; Phetsouvanh, Rattanaphone; Newton, Paul; Basnyat, Buddha; Farrar, Jeremy J.; Baker, Stephen

    2015-01-01

    Azithromycin is an effective treatment for uncomplicated infections with Salmonella enterica serovar Typhi and serovar Paratyphi A (enteric fever), but there are no clinically validated MIC and disk zone size interpretative guidelines. We studied individual patient data from three randomized controlled trials (RCTs) of antimicrobial treatment in enteric fever in Vietnam, with azithromycin used in one treatment arm, to determine the relationship between azithromycin treatment response and the azithromycin MIC of the infecting isolate. We additionally compared the azithromycin MIC and the disk susceptibility zone sizes of 1,640 S. Typhi and S. Paratyphi A clinical isolates collected from seven Asian countries. In the RCTs, 214 patients who were treated with azithromycin at a dose of 10 to 20 mg/ml for 5 to 7 days were analyzed. Treatment was successful in 195 of 214 (91%) patients, with no significant difference in response (cure rate, fever clearance time) with MICs ranging from 4 to 16 μg/ml. The proportion of Asian enteric fever isolates with an MIC of ≤16 μg/ml was 1,452/1,460 (99.5%; 95% confidence interval [CI], 98.9 to 99.7) for S. Typhi and 207/240 (86.3%; 95% CI, 81.2 to 90.3) (P azithromycin disk identified S. Typhi isolates with an MIC of ≤16 μg/ml with a sensitivity of 99.7%. An azithromycin MIC of ≤16 μg/ml or disk inhibition zone size of ≥13 mm enabled the detection of susceptible S. Typhi isolates that respond to azithromycin treatment. Further work is needed to define the response to treatment in S. Typhi isolates with an azithromycin MIC of >16 μg/ml and to determine MIC and disk breakpoints for S. Paratyphi A. PMID:25733500

  15. Pola Resistensi Salmonella typhi yang Diisolasi dari Ikan Serigala (Hoplias malabaricus terhadap Antibiotik

    Directory of Open Access Journals (Sweden)

    Waode Santa Monica

    2013-11-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE The purpose of this study was to know the resistant pattern of Salmonella typhi (S. typhi isolated from wolf fish (Hoplias malabaricus to antibiotics in Bali Safari and Marine Park. S. typhi was isolated from fecal swab collected from the illness fish. The S. typhi was confirmed with MicrogenTM GN A ID test’s method. The antibiotic sensitivity of the S. typhi was tested using streptomycin, gentamicin, penicillin, vancomycin, and chloramphenicol according to the Kirby bouer method. The results showed that the isolate of S. typhi was resistant to penicillin and vancomycin, but sensitive to streptomycin, gentamicin, and chloramphenicol. /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; text-align:justify; line-height:150%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;}

  16. Multidrug Resistant Salmonella typhi in Asymptomatic Typhoid Carriers among Food Handlers in Namakkal District, Tamil Nadu

    Directory of Open Access Journals (Sweden)

    Senthilkumar B

    2005-01-01

    Full Text Available Purpose: to screen Salmonella typhi in asymptomatic typhoid carriers and to find out drug resistance and ability of the strains to transmit drug resistance to other bacteria. Methods: Cultural characters, biochemical tests, antibiotic sensitivity test (disc diffusion, agarose gel electrophoresis, and conjugation protocols were done. Thirty five stool samples were collected from the suspected food handlers for the study. Results: Among 35 samples, (17.14% yielded a positive result. Out of these 4 (20.0% were women and 2 (13.33% were men. The isolates were tested with a number of conventional antibiotics viz, amikacin, amoxicillin, ampicillin, chloramphenicol, ciprofloxacin, co-trimaxazole, rifampicin, gentamicin, nalidixic acid, ofloxacin and tetracycline. Five isolates were having the multidrug resistant character. Four (66.66% multidrug resistant isolates were found to have plasmids, while one (16.66% multidrug resistant isolate had no plasmid and the chromosome encoded the resistance. Only one strain (16.66% showed single antibiotic resistance in the study and had no plasmid DNA. The molecular weights of the plasmids were determined and found to be 120 kb.The mechanism of spreading of drug resistance through conjugation process was analyzed. In the conjugation studies, the isolates having R+ factor showed the transfer of drug resistance through conjugation, which was determined by the development of antibiotic resistance in the recipients. Conclusion: This study shows that drug resistant strains are able to transfer genes encoding drug resistance.

  17. Activation of Salmonella Typhi-specific regulatory T cells in typhoid disease in a wild-type S. Typhi challenge model.

    Directory of Open Access Journals (Sweden)

    Monica A McArthur

    2015-05-01

    Full Text Available Salmonella Typhi (S. Typhi, the causative agent of typhoid fever, causes significant morbidity and mortality worldwide. Currently available vaccines are moderately efficacious, and identification of immunological responses associated with protection or disease will facilitate the development of improved vaccines. We investigated S. Typhi-specific modulation of activation and homing potential of circulating regulatory T cells (Treg by flow and mass cytometry using specimens obtained from a human challenge study. Peripheral blood mononuclear cells were obtained from volunteers pre- and at multiple time-points post-challenge with wild-type S. Typhi. We identified differing patterns of S. Typhi-specific modulation of the homing potential of circulating Treg between volunteers diagnosed with typhoid (TD and those who were not (No TD. TD volunteers demonstrated up-regulation of the gut homing molecule integrin α4ß7 pre-challenge, followed by a significant down-regulation post-challenge consistent with Treg homing to the gut. Additionally, S. Typhi-specific Treg from TD volunteers exhibited up-regulation of activation molecules post-challenge (e.g., HLA-DR, LFA-1. We further demonstrate that depletion of Treg results in increased S. Typhi-specific cytokine production by CD8+ TEM in vitro. These results suggest that the tissue distribution of activated Treg, their characteristics and activation status may play a pivotal role in typhoid fever, possibly through suppression of S. Typhi-specific effector T cell responses. These studies provide important novel insights into the regulation of immune responses that are likely to be critical in protection against typhoid and other enteric infectious diseases.

  18. Activation of Salmonella Typhi-specific regulatory T cells in typhoid disease in a wild-type S. Typhi challenge model.

    Science.gov (United States)

    McArthur, Monica A; Fresnay, Stephanie; Magder, Laurence S; Darton, Thomas C; Jones, Claire; Waddington, Claire S; Blohmke, Christoph J; Dougan, Gordon; Angus, Brian; Levine, Myron M; Pollard, Andrew J; Sztein, Marcelo B

    2015-05-01

    Salmonella Typhi (S. Typhi), the causative agent of typhoid fever, causes significant morbidity and mortality worldwide. Currently available vaccines are moderately efficacious, and identification of immunological responses associated with protection or disease will facilitate the development of improved vaccines. We investigated S. Typhi-specific modulation of activation and homing potential of circulating regulatory T cells (Treg) by flow and mass cytometry using specimens obtained from a human challenge study. Peripheral blood mononuclear cells were obtained from volunteers pre- and at multiple time-points post-challenge with wild-type S. Typhi. We identified differing patterns of S. Typhi-specific modulation of the homing potential of circulating Treg between volunteers diagnosed with typhoid (TD) and those who were not (No TD). TD volunteers demonstrated up-regulation of the gut homing molecule integrin α4ß7 pre-challenge, followed by a significant down-regulation post-challenge consistent with Treg homing to the gut. Additionally, S. Typhi-specific Treg from TD volunteers exhibited up-regulation of activation molecules post-challenge (e.g., HLA-DR, LFA-1). We further demonstrate that depletion of Treg results in increased S. Typhi-specific cytokine production by CD8+ TEM in vitro. These results suggest that the tissue distribution of activated Treg, their characteristics and activation status may play a pivotal role in typhoid fever, possibly through suppression of S. Typhi-specific effector T cell responses. These studies provide important novel insights into the regulation of immune responses that are likely to be critical in protection against typhoid and other enteric infectious diseases.

  19. An evaluation of purified Salmonella Typhi protein antigens for the serological diagnosis of acute typhoid fever.

    Science.gov (United States)

    Tran Vu Thieu, Nga; Trinh Van, Tan; Tran Tuan, Anh; Klemm, Elizabeth J; Nguyen Ngoc Minh, Chau; Voong Vinh, Phat; Pham Thanh, Duy; Ho Ngoc Dan, Thanh; Pham Duc, Trung; Langat, Pinky; Martin, Laura B; Galan, Jorge; Liang, Li; Felgner, Philip L; Davies, D Huw; de Jong, Hanna K; Maude, Rapeephan R; Fukushima, Masako; Wijedoru, Lalith; Ghose, Aniruddha; Samad, Rasheda; Dondorp, Arjen M; Faiz, Abul; Darton, Thomas C; Pollard, Andrew J; Thwaites, Guy E; Dougan, Gordon; Parry, Christopher M; Baker, Stephen

    2017-08-01

    The diagnosis of typhoid fever is a challenge. Aiming to develop a typhoid diagnostic we measured antibody responses against Salmonella Typhi (S. Typhi) protein antigens and the Vi polysaccharide in a cohort of Bangladeshi febrile patients. IgM against 12 purified antigens and the Vi polysaccharide was measured by ELISA in plasma from patients with confirmed typhoid fever (n = 32), other confirmed infections (n = 17), and healthy controls (n = 40). ELISAs with the most specific antigens were performed on plasma from 243 patients with undiagnosed febrile disease. IgM against the S. Typhi protein antigens correlated with each other (rho > 0.8), but not against Vi (rho 0.85, respectively. Applying a dynamic cut-off to patients with undiagnosed febrile disease suggested that 34-58% had an IgM response indicative of typhoid. We evaluated the diagnostic potential of several S. Typhi antigens; our assays give good sensitivity and specificity, but require further assessment in differing patient populations. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  20. Pengaruh Pemberian Pegagan (Centella asiatica terhadap Struktur Mikroskopis Hati Mencit Pasca Infeksi Salmonella typhi

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    Brahma Tusta Bhirawa

    2012-08-01

    Full Text Available Centella asiatica (pegagan merupakan salah satu tanaman obat yang memilikibanyak fungsi, salah satunya adalah sebagai imunostimulator, yang berarti dapatmerangsang tubuh untuk meningkatkan respon imun. Namun efek pegagan terhadapstruktur mikroskopis hati pasca diinfeksi Salmonella typhi (S.typhi belum pernahdilakukan. Penelitian ini bertujuan untuk mengetahui pengaruh ekstrak pegagan terhadapstruktur mikroskopis hati mencit yang diinfeksi S.typhi . Penelitian ini menggunakan 24ekor mencit jantan umur 8-12 minggu, yang dibagi menjadi 4 grup. P0 sebagai kontroldiberikan aquades, dan grup P1,P2, dan P3, masing-masing diberikan 125 mg/bb, 250kg/bb mg, 500 mg kg/bb. perlakuan diulang sebanyak 6 kali. Extrak pegagan diberikansecara oral selama 14 hari, pada hari ke 15 seluruh mencit diinfeksi dengan S.typhi,kemudian pada hari ke 14 semua mencit dinekropsi dan hati diambil untuk diprosespembuatan preparat dengan metode pewarnaan hematoxylin eosin (HE. Pengamatandilakukan terhadap gambaran mikroskopis berupa perdarahan, degenerasi vakuola, dannekrosis. Hasil analisis menunjukkan mencit yang tidak diberikan pegagan dengan mencityang diberikan pegagan dosis 125 mg/kg bb., memberikan hasil yang tidak berbeda nyataNamun mencit yang diberikan pegagan dosis 250 mg/kg bb, dan 500 mg/kg bb memberikangambaran mikroskopis yang berbeda nyata.

  1. An inducible and secreted eukaryote-like serine/threonine kinase of Salmonella enterica serovar Typhi promotes intracellular survival and pathogenesis.

    Science.gov (United States)

    Theeya, Nagaraja; Ta, Atri; Das, Sayan; Mandal, Rahul S; Chakrabarti, Oishee; Chakrabarti, Saikat; Ghosh, Amar N; Das, Santasabuj

    2015-02-01

    Eukaryote-like serine/threonine kinases (eSTKs) constitute an important family of bacterial virulence factors. Genome analysis had predicted putative eSTKs in Salmonella enterica serovar Typhi, although their functional characterization and the elucidation of their role in pathogenesis are still awaited. We show here that the primary sequence and secondary structure of the t4519 locus of Salmonella Typhi Ty2 have all the signatures of eukaryotic superfamily kinases. t4519 encodes a ∼39-kDa protein (T4519), which shows serine/threonine kinase activities in vitro. Recombinant T4519 (rT4519) is autophosphorylated and phosphorylates the universal substrate myelin basic protein. Infection of macrophages results in decreased viability of the mutant (Ty2Δt4519) strain, which is reversed by gene complementation. Moreover, reactive oxygen species produced by the macrophages signal to the bacteria to induce T4519, which is translocated to the host cell cytoplasm. That T4519 may target a host substrate(s) is further supported by the activation of host cellular signaling pathways and the induction of cytokines/chemokines. Finally, the role of T4519 in the pathogenesis of Salmonella Typhi is underscored by the significantly decreased mortality of mice infected with the Ty2Δt4519 strain and the fact that the competitive index of this strain for causing systemic infection is 0.25% that of the wild-type strain. This study characterizes the first eSTK of Salmonella Typhi and demonstrates its role in promoting phagosomal survival of the bacteria within macrophages, which is a key determinant of pathogenesis. This, to the best of our knowledge, is the first study to describe the essential role of eSTKs in the in vivo pathogenesis of Salmonella spp. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  2. Detection of Salmonella typhi agglutinins in sera of patients with ...

    African Journals Online (AJOL)

    Materials and Methods: Sera from 50 healthy blood donors were compared for the presence of Salmonella agglutinins in various groups of patients with other febrile illnesses using Widal test in the division of Serology and Immunology at King Khalid University Hospital, Riyadh. The patient groups of other febrile illnesses ...

  3. 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

  4. The concurrent prevalence of chloramphenicol-sensitive and multi-drug resistant Salmonella typhi in Vellore, S. India.

    OpenAIRE

    Jesudason, M. V.; John, R.; John, T. J.

    1996-01-01

    A multidrug resistant (MDR) variety of Salmonella typhi emerged as the cause of epidemic typhoid fever in some Asian countries including India, during the late 1980s. We faced the epidemic from April 1990 to the first quarter of 1993. However, during this period we continued to isolate chloramphenicol sensitive (CS) S. typhi also. The relative prevalences showed that the frequency of CS variety was unaffected by the epidemic of MDR variety. This is an unusual epidemiological pattern, which in...

  5. VNTR molecular typing of salmonella enterica serovar typhi isolates in Kathmandu valley

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    B Acharya

    2012-03-01

    Full Text Available Background: Typhoid fever continues to be a worldwide health problem, especially in developing countries. Effective epidemiological surveillance is needed to monitor the presence and spread of disease. Materials and Methods: Variable number tandem repeats (VNTR was performed for Salmonella enterica serovar typhi by multiplex-PCR in 28 Nepalese isolates of sporadic typhoid fever. Results: From all 28 total isolates, we could identify 12 VNTR profiles among the isolates, signifying multiple variants in circulation within the region. Conclusion: The VNTR-based typing assay for serovar typhi isolates can be used during an outbreak of enteric fever. The typing could eventually form the basis of an effective epidemiological surveillance system for developing rational strategies to control typhoid fever. DOI: http://dx.doi.org/10.3126/jpn.v2i3.6026 JPN 2012; 2(3: 220-223

  6. Clearance and tissue distribution of intravenously injected Salmonella typhi polysaccharide in rabbits.

    Science.gov (United States)

    Isibasi, A; Jimenez, E; Kumate, J

    1983-01-01

    The interaction of Freeman polysaccharide of Salmonella typhi with blood and tissues of rabbits was studied by radioimmunoassay. After intravenous injection of 1.0 mg of S. typhi Freeman polysaccharide, a rapid clearance phase (t1/2, 6.0 min) was followed by a slower clearance period (t1/2, 55.2 min). These results suggest first, that the distribution of whole lipopolysaccharide is a function of how the polysaccharides are handled by the host; further, that the O side chain determines how and where lipopolysaccharide is cleared from the circulatory system; and finally, that Freeman polysaccharide regulates the toxicity of lipopolysaccharide by influencing its clearance from blood. PMID:6642672

  7. Salmonella enterica serovars Typhimurium and Typhi as model organisms

    Science.gov (United States)

    Garai, Preeti; Gnanadhas, Divya Prakash; Chakravortty, Dipshikha

    2012-01-01

    The lifestyle of intracellular pathogens has always questioned the skill of a microbiologist in the context of finding the permanent cure to the diseases caused by them. The best tool utilized by these pathogens is their ability to reside inside the host cell, which enables them to easily bypass the humoral immunity of the host, such as the complement system. They further escape from the intracellular immunity, such as lysosome and inflammasome, mostly by forming a protective vacuole-bound niche derived from the host itself. Some of the most dreadful diseases are caused by these vacuolar pathogens, for example, tuberculosis by Mycobacterium or typhoid fever by Salmonella. To deal with such successful pathogens therapeutically, the knowledge of a host-pathogen interaction system becomes primarily essential, which further depends on the use of a model system. A well characterized pathogen, namely Salmonella, suits the role of a model for this purpose, which can infect a wide array of hosts causing a variety of diseases. This review focuses on various such aspects of research on Salmonella which are useful for studying the pathogenesis of other intracellular pathogens. PMID:22722237

  8. The Hd, Hj, and Hz66 flagella variants of Salmonella enterica serovar Typhi modify host responses and cellular interactions.

    Science.gov (United States)

    Schreiber, Fernanda; Kay, Sally; Frankel, Gad; Clare, Simon; Goulding, David; van de Vosse, Esther; van Dissel, Jaap T; Strugnell, Richard; Thwaites, Guy; Kingsley, Robert A; Dougan, Gordon; Baker, Stephen

    2015-01-22

    Salmonella Typhi, the causative agent of typhoid fever, is a monophyletic, human-restricted bacterium that exhibits limited phenotypic variation. S. Typhi from Indonesia are a notable exception, with circulating strains expressing diverse flagella antigens including Hj, Hd and Hz66. Hypothesizing that S. Typhi flagella plays a key role during infection, we constructed an S. Typhi fliC mutant and otherwise isogenic S. Typhi strains expressing the Hj, Hd, Hz66 flagella antigens. Phenotyping revealed differences in flagellum structure, strain motility and immunogenicity, but not in the ability of flagellated isolates to induce TLR5 activity. Invasion assays using epithelial and macrophage cell lines revealed differences in the ability of these S. Typhi derivatives to invade cells or induce cellular restructuring in the form of ruffles. Notably, the Hj variant induced substantial ruffles that were not fully dependent on the GTPases that contribute to this process. These data highlight important differences in the phenotypic properties of S. Typhi flagella variation and how they impact on the pathogenesis of S. Typhi.

  9. Interferon-γ and proliferation responses to Salmonella enterica Serotype Typhi proteins in patients with S. Typhi Bacteremia in Dhaka, Bangladesh.

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    Alaullah Sheikh

    2011-06-01

    Full Text Available Salmonella enterica serotype Typhi is a human-restricted intracellular pathogen and the cause of typhoid fever. Cellular immune responses are required to control and clear Salmonella infection. Despite this, there are limited data on cellular immune responses in humans infected with wild type S. Typhi.For this work, we used an automated approach to purify a subset of S. Typhi proteins identified in previous antibody-based immuno-affinity screens and antigens known to be expressed in vivo, including StaF-putative fimbrial protein-STY0202, StbB-fimbrial chaperone-STY0372, CsgF-involved in curli production-STY1177, CsgD- putative regulatory protein-STY1179, OppA-periplasmic oligopeptide binding protein precursor-STY1304, PagC-outer membrane invasion protein-STY1878, and conserved hypothetical protein-STY2195; we also generated and analyzed a crude membrane preparation of S. Typhi (MP. In comparison to samples collected from uninfected Bangladeshi and North American participants, we detected significant interferon-γ responses in PBMCs stimulated with MP, StaF, StbB, CsgF, CsgD, OppA, STY2195, and PagC in patients bacteremic with S. Typhi in Bangladesh. The majority of interferon-γ expressing T cells were CD4 cells, although CD8 responses also occurred. We also assessed cellular proliferation responses in bacteremic patients, and confirmed increased responses in infected individuals to MP, StaF, STY2195, and PagC in convalescent compared to acute phase samples and compared to controls. StaF is a fimbrial protein homologous to E. coli YadK, and contains a Pfam motif thought to be involved in cellular adhesion. PagC is expressed in vivo under the control of the virulence-associated PhoP-regulon required for intra-macrophage survival of Salmonella. STY2195 is a conserved hypothetical protein of unknown function.This is the first analysis of cellular immune responses to purified S. Typhi antigens in patients with typhoid fever. These results indicate

  10. Anti-biofilm efficacy of 100 MeV gold ion irradiated polycarbonate against Salmonella typhi

    Science.gov (United States)

    Joshi, R. P.; Hareesh, K.; Bankar, A.; Sanjeev, G.; Asokan, K.; Kanjilal, D.; Dahiwale, S. S.; Bhoraskar, V. N.; Dhole, S. D.

    2017-12-01

    Polycarbonate (PC) films were irradiated by 100 MeV gold (Au7+) ions and characterized to study changes in its optical, chemical, surface morphology and thermal properties. UV-Visible spectroscopic results revealed the decrease in the optical band gap of PC after ion irradiation due to chain scission mainly at the carbonyl group which is corroborated by Fourier Transform Infrared spectroscopic results. X-ray diffractogram study showed decrease in crystallinity of PC film after irradiation. Scanning electron microscopic results showed the micropores formation in PC which results in surface roughening. Differential scanning calorimetric results revealed decrease in glass transition temperature indicating the decrease in molecular weight of PC corroborated by rheometric studies. PC films irradiated by 100 MeV Au7+ ions showed increased anti-biofilm activity against the human pathogen, Salmonella typhi (S. typhi). Morphology of S. typhi was changed due to stress of Au7+ irradiated PC. Cells length was increased with increasing fluences. The average cell length, cell volume and surface area was increased significantly (P<0.05) with increasing ion fluences. Biofilm formation was inhibited ≈ 20% at lower fluence and 96% at higher fluence, which observed to be enhanced anti-biofilm activity in Au7+ irradiated PC.

  11. Non-crosslinking gold nanoprobe-LAMP for simple, colorimetric, and specific detection of Salmonella typhi

    Energy Technology Data Exchange (ETDEWEB)

    Bozorgmehr, Ali; Yazdanparast, Razieh, E-mail: ryazdan@ut.ac.ir [University of Tehran, Institute of Biochemistry and Biophysics (Iran, Islamic Republic of); Mollasalehi, Hamidreza [Shahid Beheshti University, Protein Research Center (Iran, Islamic Republic of)

    2016-12-15

    In this study, we developed a non-crosslinking gold nanoprobe loop-mediated isothermal amplification (LAMP) method for nanodiagnosis of bacterial typhoid fever source, Salmonella typhi. Therefore, a unique region in the S. typhi genomic DNA was targeted for LAMP amplification using a specific set of four precisely designed primers. Also, for specific colorimetric visualization of the amplicons, a thiolated oligonucleotide probe, complementary to the single-stranded loop region of the amplicons between F2 and F1C segments, was designed. The probe was bound to the surface of gold nanoparticles via covalent bonds. Increasing the salt concentration in the detection reaction medium led to aggregation of nanoprobes in the blank and the negative vessels in a time-dependent form. That was followed by a change in the surface plasmon resonance (SPR) leading to blue/black color that was observable by the naked eyes after about 5 min. Meanwhile, the original pink/red color was retained in the positive sample due to the large interparticle spaces and the stability against the ionic strength elevation which persisted for about 30 min. The whole process of DNA extraction, amplification, and detection took less than 2 h with a sensitivity of 20 CFU/ml. The developed gold nanoprobe-LAMP could serve as a simple, rapid, and cost-effective method for nanodiagnosis of S. typhi in point-of-need applications.

  12. Pengaruh Pemberian Pegagan (Centella asiatica terhadap Gambaran Mikroskopis Usus Halus Mencit yang Diinfeksi Salmonella typhi

    Directory of Open Access Journals (Sweden)

    I Md Chandra Arya PW

    2012-08-01

    Full Text Available Telah dilakukan penelitian mengenai pengaruh pemberian pegagan (Centella asiaticaterhadap gambaran mikroskopis usus halus mencit yang diinfeksi Salmonella typhi.Penelitian ini menggunakan sampel usus halus mencit jantan strain Balb/C yang berumur 2-3 bulan dengan berat badan berkisar antara 20-35 gram. Sebanyak 24 ekor mencit dipakaidalam penelitian, dibagi empat kelompok, yakni kelompok 1 sebagai kontrol diberikanaquades steril, kelompok 2 diberikan pegagan dengan dosis 125 mg/kg bb, kelompok 3diberikan pegagan 250 mg/kg bb, dan kelompok 4 diberikan pegagan 500 mg/kg bb.Setelah 14 hari seluruh mencit diinfeksi dengan S. typhi. Pada hari ke-15 dilakukannekropsi untuk pengambilan sampel berupa usus halus dan dibuat preparat histology,menggunakan pewarnaan Hematoxilin-Eosin. Selanjutnya dilakukan pengamatan diLaboratorium Histologi Fakultas Kedokteran Hewan, terhadap gambaran mikroskopis yangmeliputi perubahan berupa : perdarahan, infiltrasi sel radang, dan deskuamasi epitel usushalus. Metode pewarnaan menggunakan Hematoxilin-Eosin. Data yang diperoleh dianalisisdengan Uji Kruskall-Wallis. Hasil penelitian menunjukkan pemberian pegagan dengandosis 125 mg/kg bb dengan kontrol tidak berbeda nyata (p>0,05, apabila dibandingkandengan dosis pegagan250 mg/kg bb memberikan hasil yang berbeda nyata (p0,05. Hal ini berarti, pemberian ekstrak pegagan (Centella asiaticadengan dosis 250 mg/kg bb dapat mencegah perubahan gambaran mikroskopis usus halusakibat infeksi S. typhi.

  13. Non-crosslinking gold nanoprobe-LAMP for simple, colorimetric, and specific detection of Salmonella typhi

    Science.gov (United States)

    Bozorgmehr, Ali; Yazdanparast, Razieh; Mollasalehi, Hamidreza

    2016-12-01

    In this study, we developed a non-crosslinking gold nanoprobe loop-mediated isothermal amplification (LAMP) method for nanodiagnosis of bacterial typhoid fever source, Salmonella typhi. Therefore, a unique region in the S. typhi genomic DNA was targeted for LAMP amplification using a specific set of four precisely designed primers. Also, for specific colorimetric visualization of the amplicons, a thiolated oligonucleotide probe, complementary to the single-stranded loop region of the amplicons between F2 and F1C segments, was designed. The probe was bound to the surface of gold nanoparticles via covalent bonds. Increasing the salt concentration in the detection reaction medium led to aggregation of nanoprobes in the blank and the negative vessels in a time-dependent form. That was followed by a change in the surface plasmon resonance (SPR) leading to blue/black color that was observable by the naked eyes after about 5 min. Meanwhile, the original pink/red color was retained in the positive sample due to the large interparticle spaces and the stability against the ionic strength elevation which persisted for about 30 min. The whole process of DNA extraction, amplification, and detection took less than 2 h with a sensitivity of 20 CFU/ml. The developed gold nanoprobe-LAMP could serve as a simple, rapid, and cost-effective method for nanodiagnosis of S. typhi in point-of-need applications.

  14. Importance of Salmonella Typhi-Responsive CD8+ T Cell Immunity in a Human Typhoid Fever Challenge Model.

    Science.gov (United States)

    Fresnay, Stephanie; McArthur, Monica A; Magder, Laurence S; Darton, Thomas C; Jones, Claire; Waddington, Claire S; Blohmke, Christoph J; Angus, Brian; Levine, Myron M; Pollard, Andrew J; Sztein, Marcelo B

    2017-01-01

    Typhoid fever, caused by the human-restricted organism Salmonella enterica serovar Typhi (S. Typhi), constitutes a major global health problem. The development of improved attenuated vaccines is pressing, but delayed by the lack of appropriate preclinical models. Herein, we report that high levels of S. Typhi-responsive CD8+ T cells at baseline significantly correlate with an increased risk of disease in humans challenged with a high dose (~104 CFU) wild-type S. Typhi. Typhoid fever development was associated with higher multifunctional S. Typhi-responsive CD8+ T effector memory cells at baseline. Early decreases of these cells in circulation following challenge were observed in both S. Typhi-responsive integrin α4β7- and integrin α4β7+ CD8+ T effector memory (TEM) cells, suggesting their potential to home to both mucosal and extra-intestinal sites. Participants with higher baseline levels of S. Typhi-responsive CD8+ T memory cells had a higher risk of acquiring disease, but among those who acquired disease, those with a higher baseline responses took longer to develop disease. In contrast, protection against disease was associated with low or absent S. Typhi-responsive T cells at baseline and no changes in circulation following challenge. These data highlight the importance of pre-existing S. Typhi-responsive immunity in predicting clinical outcome following infection with wild-type S. Typhi and provide novel insights into the complex mechanisms involved in protective immunity to natural infection in a stringent human model with a high challenge dose. They also contribute important information on the immunological responses to be assessed in the appraisal and selection of new generation typhoid vaccines.

  15. Salmonella enterica serovar Typhi plasmid pR ST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy

    Directory of Open Access Journals (Sweden)

    Peiyan He

    Full Text Available OBJECTIVES: Plasmid pR ST98 is a hybrid resistance-virulence plasmid isolated from Salmonella enterica serovar Typhi (S. typhi. Previous studies demonstrated that pR ST98 could enhance the virulence of its host bacteria. However, the mechanism of pR ST98-increased bacterial virulence is still not fully elucidated. This study was designed to gain further insight into the roles of pR ST98 in host responses. METHODS: Human-derived macrophage-like cell line THP-1 was infected with wild-type (ST8, pR ST98-deletion (ST8-ΔpR ST98, and complemented (ST8-c-pR ST98 S. typhi strains. Macrophage autophagy was performed by extracting the membrane-unbound LC3-I protein from cells, followed by flow cytometric detection of the membrane-associated fraction of LC3-II. Intracellular bacterial growth was determined by colony-forming units (cfu assay. Macrophage cell death was measured by flow cytometry after propidium iodide (PI staining. Autophagy activator rapamycin (RAPA was added to the medium 2 h before infection to investigate the effect of autophagy on intracellular bacterial growth and macrophage cell death after S. typhi infection. RESULTS: Plasmid pR ST98 suppressed autophagy in infected macrophages and enhanced intracellular bacterial growth and S. typhi-induced macrophage cell death. Pretreatment with RAPA effectively restricted intracellular bacterial growth of ST8 and ST8-c-pR ST98, and alleviated ST8 and ST8-c-pR ST98-induced macrophage cell death, but had no significant effect on ST8-ΔpR ST98. CONCLUSIONS: Plasmid pR ST98 enhances intracellular bacterial growth and S. typhi-induced macrophage cell death by suppressing autophagy.

  16. Core-linked LPS expression of Shigella dysenteriae serotype 1 O-antigen in live Salmonella Typhi vaccine vector Ty21a: preclinical evidence of immunogenicity and protection.

    Science.gov (United States)

    Xu, De Qi; Cisar, John O; Osorio, Manuel; Wai, Tint T; Kopecko, Dennis J

    2007-08-14

    Shigella dysenteriae serotype 1 (S. dysenteriae 1) causes severe shigellosis that is typically associated with high mortality. Antibodies against Shigella serotype-specific O-polysaccharide (O-Ps) have been shown to be host protective. In this study, the rfb locus and the rfp gene with their cognate promoter regions were PCR-amplified from S. dysenteriae 1, cloned, and sequenced. Deletion analysis showed that eight rfb ORFs plus rfp are necessary for biosynthesis of this O-Ps. A tandemly-linked rfb-rfp gene cassette was cloned into low copy plasmid pGB2 to create pSd1. Avirulent Salmonella enterica serovar Typhi (S. Typhi) Ty21a harboring pSd1 synthesized S. Typhi 9, 12 LPS as well as typical core-linked S. dysenteriae 1 LPS. Animal immunization studies showed that Ty21a (pSd1) induces protective immunity against high stringency challenge with virulent S. dysenteriae 1 strain 1617. These data further demonstrate the utility of S. Typhi Ty21a as a live, bacterial vaccine delivery system for heterologous O-antigens, supporting the promise of a bifunctional oral vaccine for prevention of shigellosis and typhoid fever.

  17. Antibiotic Resistance of Salmonella enterica Serovar Typhi in Kolkata, India, and In Vitro Experiments on Effect of Combined Chemotherapy

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    Shyamapada Mandal

    2012-01-01

    Full Text Available This communication states the changing patterns of Salmonella enterica serovar Typhi (S. Typhi isolates causing enteric fever in and around Kolkata, India. Among the isolates resistance to ampicillin (A, chloramphenicol (C, cotrimoxazole (Co and tetracycline (T were plasmid mediated; the plasmid was unstable in S. Typhi, and the other enteric bacteria like Escherichia coli, Klebsiella pneumoniae and Proteus vulgaris were found to be the potential source of dissemination of such plasmids into S. Typhi. The infection with such S. Typhi strains were successfully treated with ciprofloxacin (Cp: MICs 0.0075–0.075 μg mL−1 and/or ofloxacin (Ofx: MICs 0.0125–0.075 μg mL−1, but in the later course, the S. Typhi strains, showing resistance to nalidixic acid, developed low level of resistance to Cp and Ofx, causing the treatment failure. Thus, the treatment regimen was shifted to the third generation cephalosporins like ceftriaxone (Ct and cefotaxime (Cf. Keeping in mind the anticipation of development of resistance to Ct/Cf, we prepared the treatment regimen for MDR enteric fever, based on the double-drug synergy tests in vitro; Cp-gentamycin (FICI 0.121–0.216 and Cp-trimethoprim (FICI 0.14–0.483 combinations were found effective against S. Typhi isolates having decreased sensitivity to cp (MICs: 0.5–1.25 μg mL−1.

  18. High-throughput bacterial SNP typing identifies distinct clusters of Salmonella Typhi causing typhoid in Nepalese children

    LENUS (Irish Health Repository)

    Holt, Kathryn E

    2010-05-31

    Abstract Background Salmonella Typhi (S. Typhi) causes typhoid fever, which remains an important public health issue in many developing countries. Kathmandu, the capital of Nepal, is an area of high incidence and the pediatric population appears to be at high risk of exposure and infection. Methods We recently defined the population structure of S. Typhi, using new sequencing technologies to identify nearly 2,000 single nucleotide polymorphisms (SNPs) that can be used as unequivocal phylogenetic markers. Here we have used the GoldenGate (Illumina) platform to simultaneously type 1,500 of these SNPs in 62 S. Typhi isolates causing severe typhoid in children admitted to Patan Hospital in Kathmandu. Results Eight distinct S. Typhi haplotypes were identified during the 20-month study period, with 68% of isolates belonging to a subclone of the previously defined H58 S. Typhi. This subclone was closely associated with resistance to nalidixic acid, with all isolates from this group demonstrating a resistant phenotype and harbouring the same resistance-associated SNP in GyrA (Phe83). A secondary clone, comprising 19% of isolates, was observed only during the second half of the study. Conclusions Our data demonstrate the utility of SNP typing for monitoring bacterial populations over a defined period in a single endemic setting. We provide evidence for genotype introduction and define a nalidixic acid resistant subclone of S. Typhi, which appears to be the dominant cause of severe pediatric typhoid in Kathmandu during the study period.

  19. Quantification of Salmonella Typhi in water and sediments by molecular-beacon based qPCR.

    Science.gov (United States)

    Rani, Neetika; Vajpayee, Poornima; Bhatti, Saurabh; Singh, Smriti; Shanker, Rishi; Gupta, Kailash Chand

    2014-10-01

    A molecular-beacon based qPCR assay targeting staG gene was designed for specific detection and quantification of S. Typhi and validated against water and sediment samples collected from the river Ganga, Yamuna and their confluence on two days during Mahakumbha mela 2012-2013 (a) 18 December, 2012: before six major religious holy dips (Makar Sankranti, Paush Poornima, Mauni Amavasya, Basant Panchami, Maghi Poornima and Mahashivratri) (b) 10 February, 2013: after the holy dip was taken by over 3,00,00,000 devotees led by ascetics of Hindu sects at Sangam on 'Mauni Amavasya' (the most auspicious day of ritualistic mass bathing). The assay could detect linearly lowest 1 genomic equivalent per qPCR and is highly sensitive and selective for S. Typhi detection in presence of non specific DNA from other bacterial strains including S. Paratyphi A and S. Typhimurium. It has been observed that water and sediment samples exhibit S. Typhi. The mass holy dip by devotees significantly affected the water and sediment quality by enhancing the number of S. Typhi in the study area. The qPCR developed in the study might be helpful in planning the intervention and prevention strategies for control of enteric fever outbreaks in endemic regions. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Salmonella Typhi-specific multifunctional CD8+ T cells play a dominant role in protection from typhoid fever in humans.

    Science.gov (United States)

    Fresnay, Stephanie; McArthur, Monica A; Magder, Laurence; Darton, Thomas C; Jones, Claire; Waddington, Claire S; Blohmke, Christoph J; Angus, Brian; Levine, Myron M; Pollard, Andrew J; Sztein, Marcelo B

    2016-03-01

    Typhoid fever, caused by the human-restricted organism Salmonella Typhi (S. Typhi), is a major public health problem worldwide. Development of novel vaccines remains imperative, but is hampered by an incomplete understanding of the immune responses that correlate with protection. Recently, a controlled human infection model was re-established in which volunteers received ~10(3) cfu wild-type S. Typhi (Quailes strain) orally. Twenty-one volunteers were evaluated for their cell-mediated immune (CMI) responses. Ex vivo PBMC isolated before and up to 1 year after challenge were exposed to three S. Typhi-infected targets, i.e., autologous B lymphoblastoid cell-lines (B-LCL), autologous blasts and HLA-E restricted AEH B-LCL cells. CMI responses were evaluated using 14-color multiparametric flow cytometry to detect simultaneously five intracellular cytokines/chemokines (i.e., IL-17A, IL-2, IFN-g, TNF-a and MIP-1b) and a marker of degranulation/cytotoxic activity (CD107a). Herein we provide the first evidence that S. Typhi-specific CD8+ responses correlate with clinical outcome in humans challenged with wild-type S. Typhi. Higher multifunctional S. Typhi-specific CD8+ baseline responses were associated with protection against typhoid and delayed disease onset. Moreover, following challenge, development of typhoid fever was accompanied by decreases in circulating S. Typhi-specific CD8+ T effector/memory (TEM) with gut homing potential, suggesting migration to the site(s) of infection. In contrast, protection against disease was associated with low or no changes in circulating S. Typhi-specific TEM. These studies provide novel insights into the protective immune responses against typhoid disease that will aid in selection and development of new vaccine candidates.

  1. Differential roles for pathogenicity islands SPI-13 and SPI-8 in the interaction of Salmonella Enteritidis and Salmonella Typhi with murine and human macrophages.

    Science.gov (United States)

    Espinoza, Rodrigo A; Silva-Valenzuela, Cecilia A; Amaya, Fernando A; Urrutia, Ítalo M; Contreras, Inés; Santiviago, Carlos A

    2017-02-15

    Salmonella pathogenicity island (SPI)-13 is conserved in many serovars of S. enterica, including S. Enteritidis, S. Typhimurium and S. Gallinarum. However, it is absent in typhoid serovars such as S. Typhi and Paratyphi A, which carry SPI-8 at the same genomic location. Because the interaction with macrophages is a critical step in Salmonella pathogenicity, in this study we investigated the role played by SPI-13 and SPI-8 in the interaction of S. Enteritidis and S. Typhi with cultured murine (RAW264.7) and human (THP-1) macrophages. Our results showed that SPI-13 was required for internalization of S. Enteritidis in murine but not human macrophages. On the other hand, SPI-8 was not required for the interaction of S. Typhi with human or murine macrophages. Of note, the presence of an intact copy of SPI-13 in a S. Typhi mutant carrying a deletion of SPI-8 did not improve its ability to be internalized by, or survive in human or murine macrophages. Altogether, our results point out to different roles for SPI-13 and SPI-8 during Salmonella infection. While SPI-13 contributes to the interaction of S. Enteritidis with murine macrophages, SPI-8 is not required in the interaction of S. Typhi with murine or human macrophages. We hypothesized that typhoid serovars have lost SPI-13 and maintained SPI-8 to improve their fitness during another phase of human infection.

  2. 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.

  3. Vi-CRM 197 as a new conjugate vaccine against Salmonella Typhi.

    Science.gov (United States)

    Micoli, F; Rondini, S; Pisoni, I; Proietti, D; Berti, F; Costantino, P; Rappuoli, R; Szu, S; Saul, A; Martin, L B

    2011-01-17

    An efficacious, low cost vaccine against typhoid fever, especially for young children, would make a major impact on disease burden in developing countries. The virulence capsular polysaccharide of Salmonella Typhi (Vi) coupled to recombinant mutant Pseudomonas aeruginosa exoprotein A (Vi-rEPA) has been shown to be highly efficacious. We investigated the use of carrier proteins included in infant vaccines, standardized the conjugation process and developed key assays required for routine lot release at production scale. Vi from a BSL1 organism, Citrobacter freundii, strain WR7011, was used as an alternative to Vi from S. Typhi. We showed that Vi conjugated to CRM(197), a non-toxic mutant of diphtheria toxin, widely used in commercial vaccines, was produced at high yield. Vi-CRM(197) proved immunogenic in animal studies, even without adjuvant. Thus, Vi-CRM(197) appears to be a suitable candidate for the development of a commercially viable, effective typhoid vaccine for developing countries. Copyright © 2010 Elsevier Ltd. All rights reserved.

  4. Pengaruh Pemberian Pegagan (Centella asiatica terhadap Gambaran Mikroskopis Limpa Mencit yang Diinfeksi Salmonella typhi

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    I Gede Oka Budiawan

    2013-02-01

    Full Text Available Penelitian ini bertujuan untuk mengetahui pengaruh pemberian ekstrak pegagan terhadap gambaran histologis limpa mencit yang diinfeksi Salmonella typhi. Mencit sebanyak 24 ekor dibagi empat kelompok, yakni kelompok I (P0 sebagai kontrol diberikan aquades steril, kelompok II (P1 diberikan pegagan dengan dosis 125 mg/kg bb, kelompok III (P2 diberikan pegagan 250 mg/kg bb, dan kelompok IV (P3 diberikan pegagan 500 mg/kg bb. Setiap perlakuan diulang sebanyak 6 kali. Pegagan di berikan setiap hari selama 14 hari. Setelah 14 hari seluruh mencit diinfeksi S.typhi. Pada hari ke-15 dilakukan nekropsi untuk pengambilan sampel berupa limpa dan dibuat preparat histologi. Pengamatan preparat dilakukan di Laboratorium Histologi, meliputi persentase nekrosis. Metote pewarnaan menggunakan Haematoxylin Eosin (HE. Data yang diperoleh dianalisis dengan Uji Kruskal-Wallis. Hasil analisis menunjukan gambaran  mikroskopis limpa mencit yang diberikan pegagan dosis 500 mg/kg bb berbeda nyata (P<0,05 dengan limpa mencit yang diberikan pegagan dosis 125 mg/kg bb, 250 mg/kg bb dan mencit yang tidak diberikan pegagan.

  5. Pengaruh Pemberian Pegagan (Centella asiatica Terhadap Gambaran Mikroskopis Ginjal Mencit yang Diinfeksi Salmonella typhi

    Directory of Open Access Journals (Sweden)

    Komang Ariya Hendrayana

    2013-02-01

    Full Text Available Telah dilakukan penelitian mengenai pengaruh pemberian pegagan (Centella asiatica terhadap gambaran mikroskopis ginjal mencit yang telah diinfeksi  Salmonella typhi. Penelitian menggunakan 24 ekor  mencit jantan berumur 8 – 12 minggu, dengan berat badan 20 – 35 gram, dibagi menjadi empat perlakuan dengan enam kali ulangan, yaitu kelompok P0 dengan pemberian aquades steril, kelompok P1 dengan pemberian pegagan 125 mg/kg bb, kelompok P2 dengan pemberian pegagan 250 mg/kg bb, dan kelompok P3 dengan pemberian 500 mg/kg bb. Setelah 14 hari seluruh mencit diinfeksi dengan S. typhi. Pada hari ke 15 dilakukan nekropsi untuk pengambilan sampel berupa ginjal dan dibuat preparat histologi. Selanjutnya dilakukan pengamatan terhadap gambaran mikroskopis yang meliputi perubahan : infiltrasi sel radang, perdarahan, degenerasi vakuola dan nekrosis. Metode pewarnaan menggunakan Haematoxylin dan Eosin (HE. Hasil penelitian menunjukkan perlakuan P0 berbeda nyata dengan P1, P2, dan P3. Demikian juga dengan derajat kerusakan ginjal tersebut. Semakin tinggi dosis pegagan yang diberikan semakin berpengaruh terhadap perbaikan struktur mikroskopis ginjal.

  6. Enhancing chloramphenicol and trimethoprim in vitro activity by Ocimum sanctum Linn. (Lamiaceae) leaf extract against Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Mandal, Shyamapada; Mandal, Manisha Deb; Pal, Nishith Kumar

    2012-03-01

    To evaluate the antibacterial activity of Ocimum sanctum (O. sanctum) leaf extract, alone, and in combination with chloramphenicol (C) and trimethoprim (Tm) against Salmonella enterica serovar Typhi (S. typhi). The antibacterial activity of ethanolic extract of tulsi, O. sanctum, leaf (TLE; 500 μg) for 23 S. typhi isolates was determined following agar diffusion. The C (30 μg) and Tm (5 μg) activity alone and in combination with TLE (250 μg) was determined by disk diffusion. The zone diameter of inhibition (ZDI) for the agents was recorded, and growth inhibitory indices (GIIs) were calculated. The S. typhi isolates (n=23), which were resistant to both C (ZDI 6 mm) and Tm (ZDI 6 mm), had TLE (500 μg) ZDIs 16-24 mm. The ZDIs of C and Tm were increased up to 15-21 mm and 17-23 mm, respectively, when TLE (250 μg) was added to the C and Tm discs. The GIIs ranged 0.789-1.235 and 0.894-1.352, due to combined activity against S. typhi isolates, of C and TLE and Tm and TLE, respectively. The data suggest that TLE, in combination with C and Tm, had synergistic activity for S. typhi isolates, and hence O. sanctum is potential in combating S. typhi drug resistance, as well promising in the development of non-antibiotic drug for S. typhi infection. Copyright © 2012 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  7. Generation of a murine monoclonal antibody to capsular polysaccharide Vi from Salmonella Typhi

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    Fátima Reyes-López

    2015-11-01

    Full Text Available The conventional hybridoma technology has enabled the development of monoclonal antibodies (Mabs against many antigens. Mabs have several applications in the field of basic research, diagnosis, immunotherapy and vaccine manufacturing processes. Mabs-producing hybridomas against the capsular polysaccharide from Salmonella Typhi were obtained, after intraperitoneal immunization of BALB/c mice with 10 µg of capsular polysaccharide Vi conjugated to diphtheria toxoid, and subsequent fusion of lymphocytes isolated of the spleen and myeloma cells SP2/O. A Mab was selected, partially characterized, and named as 4G3E11. The isotype of this Mab was IgG1. It was proved by means of a sandwich ELISA that the 4G3E11 Mab reacts with different concentrations of polysaccharide in samples of the vax-TyVi® vaccine. The Mab obtained in this research could be useful as reagent for the detection and quantitation of polysaccharide Vi in typhoid vaccines.

  8. POTENSI Salmonella typhi YANG DILEMAHKAN DENGAN SINAR ULTRAVIOLET SEBAGAI VAKSIN ALTERNATIF

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    Andreas Putro Ragil Santoso

    2016-03-01

    Full Text Available Salmonella typhi is a Gram-negative intracellular bacterium and causes typhoid fever in humans. The success rate of Berma Vivotif Ty21a vaccine in Indonesia is only 33-66%, while in other countries have been reached up to 100%. The research was conducted in order to determine the potency of local isolate bacteria to stimulate the immune response and the impact of different exposure frequencies on the immune response and the different immune response time when administered by UV-inactivated vaccine.. The results showed that the antibody titer of local isolates irradiated by UV light 10x was 88.76 ± 33.06 IU/mL at week 4 with the lowest antibody titer values about 11.15 ± 9.18 IU/mL was found in the negative control.

  9. Molecular typing of Salmonella typhi strains from Dhaka (Bangladesh) and development of DNA probes identifying plasmid-encoded multidrug-resistant isolates

    NARCIS (Netherlands)

    P.W.M. Hermans (Peter); S.K. Saha; W.J. van Leeuwen (Wibeke); H.A. Verbrugh (Henri); A.F. van Belkum (Alex); W.H.F. Goessens (Wil)

    1996-01-01

    textabstractSeventy-eight Salmonella typhi strains isolated in 1994 and 1995 from patients living in Dhaka, Bangladesh, were subjected to phage typing, ribotyping, IS200 fingerprinting, and PCR fingerprinting. The collection displayed a high degree of genetic

  10. 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

    2014-09-02

    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. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Antibacterial effect of mango (Mangifera indica Linn.) leaf extract against antibiotic sensitive and multi-drug resistant Salmonella typhi.

    Science.gov (United States)

    Hannan, Abdul; Asghar, Samra; Naeem, Tahir; Ikram Ullah, Muhammad; Ahmed, Ijaz; Aneela, Syeda; Hussain, Shabbir

    2013-07-01

    Alternative herbal medicine has been used to treat various infections from centuries. Natural plants contain phytoconstituents having similar chemical properties as of synthetic antibiotics. Typhoid fever is a serious infection and failure of its treatment emerged multi-drug resistant (MDR) bugs of Salmonella typhi. Due to multiple and repeated issues with antibiotics efficacy, it became essential to evaluate biological properties of plants from different geographical origins. Mango leaves have been Reported for various medicinal effects like antioxidant, antimicrobial, antihelminthic, antidiabetic and antiallergic etc. Objective of present study was to investigate anti-typhoid properties of acetone mango leaf extract (AMLE) against antibiotic sensitive and MDR S. typhi isolates. A total of 50 isolates of S. typhi including MDR (n=30) and antibiotic sensitive (n=20) were investigated. Staphylococcus aureus (ATCC 25923) and Salmonella typhimurium (ATCC14028) were used as quality control strains. AMLE was prepared and its antibacterial activity was evaluated by agar well diffusion screening method and minimum inhibitory concentration (MIC), by agar dilution technique. Zone of inhibition (mm) of AMLE against MDR and antibiotic sensitive isolates was 18±1.5mm (Mean±S.D). Zone of S. aureus (ATCC 25923) and S. typhimurium (ATCC14028) was 20±1.5mm (Mean±S.D). MIC of AMLE was Reported in range from 10-50 mg/ml. The present study described the inhibitory effects of mango leaves against S. typhi.

  12. [A case with fever of unknown origin during treatment for malaria: multi-drug resistant Salmonella typhi infection].

    Science.gov (United States)

    Akin, Kemal; Yavuzdemir, Sükran; Cesur, Salih; Levent, Belkis; Esen, Berrin; Willke, Ayşe

    2005-01-01

    Typhoid fever is an acute infectious disease caused by Salmonella serotype Typhi, leading to endemic or epidemic outbreaks in tropical/ subtropical countries (especially in India, Southeast Asia, Central and South Africa). In this report, a 27 years old male patient with malaria has been presented. The patient was diagnosed to have malaria while working in Afghanistan, and received malaria treatment since one month. He admitted to our hospital because of still continuing high fever, and other complaints (weight loss, night sweats, weakness, anorexia). His fever was 39.5 degrees C at admission, and blood smears were negative for Plasmodium sp. On the third day of admission, rose spots were detected on the skin of the abdomen and chest, and group agglutination tests gave positive results for S. Typhi O (titer: 1/800), and S. Typhi H (titer: 1/3200). Blood cultures revealed growth of Salmonella enterica serotip Typhi. The isolate was found to be resistant to ampicillin, chloramphenicol, tetracyclin and trimethoprimsulfamethoxazole, and sensitive to ciprofloxacin. The patient was treated successfully with ciprofloxacin for 14 days.

  13. Whole Genome Sequence Analysis of Salmonella Typhi Isolated in Thailand before and after the Introduction of a National Immunization Program.

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    Zoe A Dyson

    2017-01-01

    Full Text Available Vaccines against Salmonella Typhi, the causative agent of typhoid fever, are commonly used by travellers, however, there are few examples of national immunization programs in endemic areas. There is therefore a paucity of data on the impact of typhoid immunization programs on localised populations of S. Typhi. Here we have used whole genome sequencing (WGS to characterise 44 historical bacterial isolates collected before and after a national typhoid immunization program that was implemented in Thailand in 1977 in response to a large outbreak; the program was highly effective in reducing typhoid case numbers. Thai isolates were highly diverse, including 10 distinct phylogenetic lineages or genotypes. Novel prophage and plasmids were also detected, including examples that were previously only reported in Shigella sonnei and Escherichia coli. The majority of S. Typhi genotypes observed prior to the immunization program were not observed following it. Post-vaccine era isolates were more closely related to S. Typhi isolated from neighbouring countries than to earlier Thai isolates, providing no evidence for the local persistence of endemic S. Typhi following the national immunization program. Rather, later cases of typhoid appeared to be caused by the occasional importation of common genotypes from neighbouring Vietnam, Laos, and Cambodia. These data show the value of WGS in understanding the impacts of vaccination on pathogen populations and provide support for the proposal that large-scale typhoid immunization programs in endemic areas could result in lasting local disease elimination, although larger prospective studies are needed to test this directly.

  14. The Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for the Rapid Detection of Salmonella enterica serovar Typhi.

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    Fenxia Fan

    Full Text Available Typhoid fever remains a public health threat in many countries. A positive result in traditional culture is a gold-standard for typhoid diagnosis, but this method is time consuming and not sensitive enough for detection of samples containing a low copy number of the target organism. The availability of the loop-mediated isothermal amplification (LAMP assay, which offers high speed and simplicity in detection of specific targets, has vastly improved the diagnosis of numerous infectious diseases. However, little research efforts have been made on utilizing this approach for diagnosis of Salmonella enterica serovar Typhi by targeting a single and specific gene. In this study, a LAMP assay for rapid detection of S. Typhi based on a novel marker gene, termed STY2879-LAMP, was established and evaluated with real-time PCR (RT-PCR. The specificity tests showed that STY2879 could be amplified in all S. Typhi strains isolated in different years and regions in China, whereas no amplification was observable in non-typhoidal strains covering 34 Salmonella serotypes and other pathogens causing febrile illness. The detection limit of STY2879-LAMP for S. Typhi was 15 copies/reaction in reference plasmids, 200 CFU/g with simple heat-treatment of DNA extracted from simulated stool samples and 20 CFU/ml with DNA extracted from simulated blood samples, which was 10 fold more sensitive than the parallel RT-PCR control experiment. Furthermore, the sensitivity of STY2879-LAMP and RT-PCR combining the traditional culture enrichment method for simulated stool and blood spiked with lower S. Typhi count during the 10 h enrichment time was also determined. In comparison with LAMP, the positive reaction time for RT-PCR required additional 2-3 h enrichment time for either simulated stool or blood specimens. Therefore, STY2879-LAMP is of practical value in the clinical settings and has a good potential for application in developing regions due to its easy-to-use protocol.

  15. Detection of blaCTX-M Extended Spectrum Beta-lactamase Producing Salmonella enterica Serotype Typhi in a Tertiary Care Centre.

    Science.gov (United States)

    Ramachandran, Aishwarya; Shanthi, Mariappan; Sekar, Uma

    2017-09-01

    Infections caused by Salmonella are an important public health threat in tropical and subtropical countries. Due to the emergence of resistance to ampicillin, chloramphenicol and trimethoprim/sulfamethoxazole (multidrug resistant salmonellae) in the late 1980s, fluoroquinolones and extended spectrum cephalosporins became the drugs of choice. Resistance to cefotaxime and ceftriaxone due to the production of Extended Spectrum Beta-Lactamase (ESBL) and reduced susceptibility to ciprofloxacin have emerged resulting in treatment failure. The Cefotaximase (CTX-M) type ESBLs are the most widespread beta lactamase among Enterobacteriaceae including salmonellae. To detect the presence of blaCTX-M in salmonellae causing human infections. Detection of qnr genes to identify the coexistence of blaCTX-M and qnr gene. The study included 103 consecutive, non-repetitive salmonellae isolated from clinical specimens obtained from July 2015- June 2016 which were identified up to species level by conventional/automated methods. Susceptibility to various classes of antimicrobial agents was determined by disc diffusion method. Minimum Inhibitory Concentration (MIC) to cefotaxime and ceftriaxone was determined by agar dilution method. The results were interpreted in accordance with Clinical & Laboratory Standard Institute (CLSI) (guidelines 2015. Detection of the ESBL phenotype was performed by the combined disk method. Polymerase Chain Reaction (PCR) amplification of all isolates was performed using group specific primers to characterize the presence of blaCTX-M, qnrA, qnrB and qnrS. Of the 103 study isolates two isolates of Salmonella typhi were resistant to cefotaxime and ceftriaxone and had a MIC of 128μg/ml. PCR amplification and sequencing detected the presence of blaCTX-M-15 in these two isolates. These two isolates exhibited resistance to ciprofloxacin in vitro but qnr gene was not detected in these isolates. Resistance to third generation cephalosporins among salmonellae is a

  16. Cross-Reactivity of Rapid Salmonella Typhi IgM Immunoassay in Dengue Fever Without Co-Existing Infection.

    Science.gov (United States)

    Bhatti, Adnan Bashir; Ali, Farhan; Satti, Siddique Akbar

    2015-12-04

    Dengue fever is endemic in developing nations worldwide with as many as 500,000 annual cases of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). A prompt and accurate diagnosis early in the disease course is essential for prompt identification and treatment of severe complications of the dengue virus infection (DVI). We identified cross-reactivity of a rapid IgM test for typhoid fever in patients with febrile illnesses that were determined to be due to dengue virus. All patients with documented DVI during a recent epidemic in Pakistan also underwent diagnostic testing for Salmonella enterica serovar Typhi. The diagnosis of DVI was made based on clinical findings and the positive results for dengue non-structural protein 1 antigen (NS1Ag) and/or dengue IgM antibody (anti-D IgM) during the acute phase of febrile illness. Patients with positive test results for Salmonella typhi (S. Typhi) IgM also had their blood cultures done. In the group of 322 patients with clinical and serological evidence of DVI, 107 also tested positive for S. Typhi IgM. Blood cultures were negative for S. Typhi bacteria in all patients. Principal disease features included fever, headache, myalgia, retro-orbital pain, and a rash accompanied by thrombocytopenia and leukopenia. Comparisons of clinical and routine laboratory findings between the S. Typhi-positive and negative groups showed no significant differences. Patients testing positive for both NS1Ag and anti-D IgM were significantly more likely to test positive for S. Typhi IgM, even in the absence of typhoid fever. No routine antibiotics were used and all patients survived. One-third of a large group of patients with primary DVI also demonstrated false positive results for typhoid fever. Cross-reactivity of a rapid immunoassay for typhoid fever has not been previously reported in DVI or any other flavivirus infections. Until these findings can be further evaluated, clinicians should be cautious in

  17. Factors Associated with Encephalopathy in Patients with Salmonella enterica Serotype Typhi Bacteremia Presenting to a Diarrheal Hospital in Dhaka, Bangladesh

    Science.gov (United States)

    Leung, Daniel T.; Bogetz, Jori; Itoh, Megumi; Ganapathi, Lakshmi; Pietroni, Mark A. C.; Ryan, Edward T.; Chisti, Mohammod Jobayer

    2012-01-01

    To characterize clinical correlates of typhoid fever-associated encephalopathy, we performed a retrospective chart review of patients with Salmonella enterica serotype Typhi bacteremia who were hospitalized at the International Centre for Diarrhoeal Disease Research, Bangladesh, from February of 2009 to June of 2011. Of 207 patients bacteremic with Salmonella Typhi who were ≥ 5 years of age, we identified 43 (21%) patients with encephalopathy. Univariate analysis revealed that patients with encephalopathy more often presented at ages of 10–24 years and had severe dehydration, low oxygen saturation, high respiratory rate, low leukocyte count, low platelet count, and Widal flagellar H agglutinin (TH) titer ≥ 1:640 compared with typhoid patients without encephalopathy. Multivariate analysis using logistic regression showed that age, dehydration, leukocyte count, and Widal TH titer were independently associated with encephalopathy. Our findings suggest that age, severity of disease, and immune responses are associated with encephalopathy during Salmonella Typhi bacteremia, perhaps reflecting the impact of prominent inflammatory responses. PMID:22492156

  18. Isolasi karakterisasi dan pengelompokan strain Salmonella typhi asal Kabupaten Sumba Barat Daya Nusa Tenggara Timur berdasarkan sifat-sifat fenotip

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    Charis Amarantini

    2012-02-01

    Full Text Available Typhoid fever is highly endemic in the South-West Sumba Regency, East Nusa Tenggara. The incidence rate of the diseases is high estimated at 725/100,000. It is an acute systemic infection caused by Salmonella typhi. The clinical symptoms of the disease are extremely diverse, starting from the mild form to severe ones with the most feared complication being perforation within the small intestine. Therefore, it is important to perform isolation, characterization, and grouping of S. typhi strains from the blood culture in order to determined definitely diagnosis and the different phenotypic characteristics in the community. Isolation was done in selective and differential media: BacT/ALERT FA culture media, Selenite Cystine Broth, Chromocult Coliform Agar, MacConkey Agar, andSalmonella Shigella Agar. The typical colony of Salmonella was confirmed on Triple Sugar Iron Agar, Urea agar, and L-Lysinedecarboxylation media. Phenotypic characteristics of all isolates were identified using API 20E and API 50CHE diagnostics. Based on biochemical characteristics the result showed that 18 strains obtained from different geographical origins were diverse. Four strains have similarity value 100% while the remained strains have similarity value 86.3-98.4%. All of the strains were categorized in the species of S. typhi.

  19. Inhibition of Salmonella typhi growth using extremely low frequency electromagnetic (ELF-EM) waves at resonance frequency.

    Science.gov (United States)

    Fadel, M A; Mohamed, S A; Abdelbacki, A M; El-Sharkawy, A H

    2014-08-01

    Typhoid is a serious disease difficult to be treated with conventional drugs. The aim of this study was to demonstrate a new method for the control of Salmonella typhi growth, through the interference with the bioelectric signals generated from the microbe during cell division by extremely low frequency electromagnetic waves (ELF-EMW-ELF-EM) at resonance frequency. Isolated Salmonella typhi was subjected to square amplitude modulated waves (QAMW) with different modulation frequencies from two generators with constant carrier frequency of 10 MHz, amplitude of 10 Vpp, modulating depth ± 2 Vpp and constant field strength of 200 V m(-1) at 37°C. Both the control and exposed samples were incubated at the same conditions during the experiment. The results showed that there was highly significant inhibition effect for Salm. typhi exposed to 0·8 Hz QAMW for a single exposure for 75 min. Dielectric relaxation, TEM and DNA results indicated highly significant changes in the molecular structure of the DNA and cellular membrane resulting from the exposure to the inhibiting EM waves. It was concluded that finding out the inhibiting resonance frequency of ELF-EM waves that deteriorates Salm. typhi growth will be promising method for the treatment of Salm. typhi infection either in vivo or in vitro. This new non-invasive technique for treatment of bacterial infections is of considerable interest for the use in medical and biotechnological applications. © 2014 The Society for Applied Microbiology.

  20. The Prevalence and Antibiotic Susceptibility Pattern of Salmonella typhi among Patients Attending a Military Hospital in Minna, Nigeria

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    N. U. Adabara

    2012-01-01

    Full Text Available The threat to human health posed by antibiotic-resistant bacterial pathogens is of growing concern to medical practice. This study investigated the antibiotic sensitivity pattern of Salmonella typhi isolated from blood specimen. One hundred blood samples were collected from suspected typhoid fever patients in 31 Artillery Brigade Medical Centre, Minna, and were analyzed for S. typhi while antibiotic sensitivity testing was done Kirby-Bauer method. Sixty (60.0% samples out of the total 100 were positive for bacterial growth. The organisms isolated 2 include Salmonella typhi; 45 (75.0%, Shigella; 6 (10.0%, E. coli; 3 (5.0%, Klebsiella; 3 (5.0%, Enterobacter; 2 (3.3%, and Citrobacter; 1 (1.7%. Result of the sensitivity test showed that the isolates were resistant to all the antibiotics; ceftriaxone, cefuroxime, amoxicillin, ampicillin, ciprofloxacin, and augmentin, which are the drug of choice routinely used in the study area for the treatment of typhoid fever. They were however sensitive to chloramphenicol and ofloxacin, which, unfortunately, are not used in this study area for the treatment of typhoid fever. There appear to be multiple drug resistant (MDR strain of S. typhi in the study area. These may be as a result of overdependence or uncontrolled use of the few available antibiotics and/or inaccurate or inconclusive diagnosis resulting in the development and spread of resistant strains of S. typhi. The study, therefore, highlights the need for a strong collaboration between the physicians and the laboratory in the choice of antibiotics for the treatment of bacterial diseases in order to discourage the development of resistant strain of bacterial pathogen.

  1. Priming of Salmonella enterica serovar typhi-specific CD8(+ T cells by suicide dendritic cell cross-presentation in humans.

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    Rosângela Salerno-Goncalves

    2009-06-01

    Full Text Available The emergence of antibiotic-resistant strains of Salmonella enterica serovar Typhi (S. Typhi, the etiologic agent of typhoid fever, has aggravated an already important public health problem and added new urgency to the development of more effective typhoid vaccines. To this end it is critical to better understand the induction of immunity to S. Typhi. CD8(+ T cells are likely to play an important role in host defense against S. Typhi by several effector mechanisms, including killing of infected cells and IFN-gamma secretion. However, how S. Typhi regulates the development of specific CD8(+ responses in humans remains unclear. Recent studies in mice have shown that dendritic cells (DC can either directly (upon uptake and processing of Salmonella or indirectly (by bystander mechanisms elicit Salmonella-specific CD8(+ T cells.We report here that upon infection with live S. Typhi, human DC produced high levels of pro-inflammatory cytokines IL-6, IL-8 and TNF-alpha, but low levels of IL-12 p70 and IFN-gamma. In contrast, DC co-cultured with S. Typhi-infected cells, through suicide cross-presentation, uptake S. Typhi-infected human cells and release high levels of IFN-gamma and IL-12p70, leading to the subsequent presentation of bacterial antigens and triggering the induction of memory T cells, mostly CD3(+CD8(+CD45RA(-CD62L(- effector/memory T cells.This study is the first to demonstrate the effect of S. Typhi on human DC maturation and on their ability to prime CD8(+ cells and highlights the significance of these phenomena in eliciting adaptive immunity to S. Typhi.

  2. Anthrax protective antigen delivered by Salmonella enterica serovar Typhi Ty21a protects mice from a lethal anthrax spore challenge.

    Science.gov (United States)

    Osorio, Manuel; Wu, Yanping; Singh, Sunil; Merkel, Tod J; Bhattacharyya, Siba; Blake, Milan S; Kopecko, Dennis J

    2009-04-01

    Bacillus anthracis, the etiological agent of anthrax disease, is a proven weapon of bioterrorism. Currently, the only licensed vaccine against anthrax in the United States is AVA Biothrax, which, although efficacious, suffers from several limitations. This vaccine requires six injectable doses over 18 months to stimulate protective immunity, requires a cold chain for storage, and in many cases has been associated with adverse effects. In this study, we modified the B. anthracis protective antigen (PA) gene for optimal expression and stability, linked it to an inducible promoter for maximal expression in the host, and fused it to the secretion signal of the Escherichia coli alpha-hemolysin protein (HlyA) on a low-copy-number plasmid. This plasmid was introduced into the licensed typhoid vaccine strain, Salmonella enterica serovar Typhi strain Ty21a, and was found to be genetically stable. Immunization of mice with three vaccine doses elicited a strong PA-specific serum immunoglobulin G response with a geometric mean titer of 30,000 (range, 5,800 to 157,000) and lethal-toxin-neutralizing titers greater than 16,000. Vaccinated mice demonstrated 100% protection against a lethal intranasal challenge with aerosolized spores of B. anthracis 7702. The ultimate goal is a temperature-stable, safe, oral human vaccine against anthrax infection that can be self-administered in a few doses over a short period of time.

  3. Recognition of nomenclatural standing of Salmonella typhi (Approved Lists 1980), Salmonella enteritidis (Approved Lists 1980) and Salmonella typhimurium (Approved Lists 1980), and conservation of the specific epithets enteritidis and typhimurium. Request for an opinion.

    Science.gov (United States)

    Ezaki, T; Kawamura, Y; Yabuuchi, E

    2000-03-01

    In 1994, the Judicial Commission of the ICSB announced that Le Minor and Popoff's Request for an Opinion in 1987 to designate Salmonella enterica sp. nov., nom. rev. as the type and only species of the genus Salmonella was denied. Thus, the current species of the genus Salmonella are Salmonella typhimurium, Salmonella enteritidis, Salmonella typhi, Salmonella choleraesuis (including six subspecies) and Salmonella bongori, with the type species, S. choleraesuis (Smith 1894) Weldin 1927 (Approved Lists 1980). Because the decision of the Judicial Commission about the request by Le Minor in 1987 was suspended for 7 years, the non-validated name 'S. enterica' has been used among microbiologists and has caused confusion in the nomenclature of Salmonella. In order to overcome such confusion, and because of their importance as human pathogens, we herein propose to recognize the nomenclatural status of S. typhi, S. enteritidis and S. typhimurium as species and request to issue an Opinion to conserve the specific epithets enteritidis and typhimurium in the species names Salmonella enteritidis and Salmonella typhimurium.

  4. Clinical application of a dot blot test for diagnosis of enteric fever due to Salmonella enterica serovar typhi in patients with typhoid fever from Colombia and Peru.

    Science.gov (United States)

    Cardona-Castro, N; Gotuzzo, E; Rodriguez, M; Guerra, H

    2000-03-01

    Clinical application of a dot blot test to detect immunoglobulin G (IgG) (88% sensitivity and specificity) and IgM (12.1% sensitivity and 97% specificity) against flagellar antigen from Salmonella enterica serovar Typhi was performed in Peruvian and Colombian patients with typhoid fever. This test can be used as a good predictor of serovar Typhi infection in regions lacking laboratory facilities and in field studies.

  5. The Ecological Dynamics of Fecal Contamination and Salmonella Typhi and Salmonella Paratyphi A in Municipal Kathmandu Drinking Water

    Science.gov (United States)

    Walker, Alan W.; Thompson, Corinne N.; Torres, Andres; Dongol, Sabina; Tran Vu Thieu, Nga; Pham Thanh, Duy; Tran Thi Ngoc, Dung; Voong Vinh, Phat; Singer, Andrew C.; Parkhill, Julian; Thwaites, Guy; Basnyat, Buddha; Ferguson, Neil; Baker, Stephen

    2016-01-01

    One of the UN sustainable development goals is to achieve universal access to safe and affordable drinking water by 2030. It is locations like Kathmandu, Nepal, a densely populated city in South Asia with endemic typhoid fever, where this goal is most pertinent. Aiming to understand the public health implications of water quality in Kathmandu we subjected weekly water samples from 10 sources for one year to a range of chemical and bacteriological analyses. We additionally aimed to detect the etiological agents of typhoid fever and longitudinally assess microbial diversity by 16S rRNA gene surveying. We found that the majority of water sources exhibited chemical and bacterial contamination exceeding WHO guidelines. Further analysis of the chemical and bacterial data indicated site-specific pollution, symptomatic of highly localized fecal contamination. Rainfall was found to be a key driver of this fecal contamination, correlating with nitrates and evidence of S. Typhi and S. Paratyphi A, for which DNA was detectable in 333 (77%) and 303 (70%) of 432 water samples, respectively. 16S rRNA gene surveying outlined a spectrum of fecal bacteria in the contaminated water, forming complex communities again displaying location-specific temporal signatures. Our data signify that the municipal water in Kathmandu is a predominant vehicle for the transmission of S. Typhi and S. Paratyphi A. This study represents the first extensive spatiotemporal investigation of water pollution in an endemic typhoid fever setting and implicates highly localized human waste as the major contributor to poor water quality in the Kathmandu Valley. PMID:26735696

  6. Development of a multiplex polymerase chain reaction protocol for the simultaneous detection of Salmonella enterica serovar Typhi and Class 1 integron

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    Juthika Mandal

    2014-09-01

    Full Text Available Objective: To develop a multiplex polymerase chain reaction (PCR protocol for the simultaneous detection of Salmonella enterica serovar Typhi (S. Typhi and Class 1 integron, so as to aid rapid diagnosis of S. Typhi cases and help in the selection of treatment options based on the presence of the Class 1 integron that can carry resistance cassettes to a range of antibiotics. Methods: PCR for amplification of specific regions was done using fliC-d and intl primers and agarose gel electrophoresis was used for resolution of PCR products. Results: The fliC-d primer (S. Typhi specific amplified a 587 bp region and the intl primer (Class 1 integron specific amplified two bands approximately 500 and 550 bps. The developed method was specific for S. Typhi and did not amplify any products with Salmonella enterica serovar Typhimurium ATCC 14028, Salmonella enterica serovar Paratyphi and Escherichia coli O157:H7. Conclusions: The developed multiplex PCR protocol can be used for rapid diagnosis and aid in proper treatment strategies for patients infected with S. Typhi.

  7. Development of an Acid-Resistant Salmonella Typhi Ty21a Attenuated Vector For Improved Oral Vaccine Delivery.

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    Madushini N Dharmasena

    Full Text Available The licensed oral, live-attenuated bacterial vaccine for typhoid fever, Salmonella enterica serovar Typhi strain Ty21a, has also been utilized as a vaccine delivery platform for expression of diverse foreign antigens that stimulate protection against shigellosis, anthrax, plague, or human papilloma virus. However, Ty21a is acid-labile and, for effective oral immunization, stomach acidity has to be either neutralized with buffer or by-passed with Ty21a in an enteric-coated capsule (ECC. Several studies have shown that efficacy is reduced when Ty21a is administered in an ECC versus as a buffered liquid formulation, the former limiting exposure to GI tract lymphoid tissues. However, the ECC was selected as a more practical delivery format for both packaging/shipping and vaccine administration ease. We have sought to increase Ty21a acid-resistance to allow for removal from the ECC and immune enhancement. To improve Ty21a acid-resistance, glutamate-dependent acid resistance genes (GAD; responsible for Shigella spp. survival at very low pH were cloned on a multi-copy plasmid (pGad under a controllable arabinose-inducible promoter. pGad enhanced acid survival of Ty21a by 5 logs after 3 hours at pH 2.5, when cells were pre-grown in arabinose and under conditions that promote an acid-tolerance response (ATR. For genetically 100% stable expression, we inserted the gad genes into the Ty21a chromosome, using a method that allowed for subsequent removal of a selectable antibiotic resistance marker. Further, both bacterial growth curves and survival assays in cultured human monocytes/macrophages suggest that neither the genetic methods employed nor the resulting acid-resistance conferred by expression of the Gad proteins in Ty21a had any effect on the existing attenuation of this vaccine strain.

  8. Development of an Acid-Resistant Salmonella Typhi Ty21a Attenuated Vector For Improved Oral Vaccine Delivery.

    Science.gov (United States)

    Dharmasena, Madushini N; Feuille, Catherine M; Starke, Carly Elizabeth C; Bhagwat, Arvind A; Stibitz, Scott; Kopecko, Dennis J

    The licensed oral, live-attenuated bacterial vaccine for typhoid fever, Salmonella enterica serovar Typhi strain Ty21a, has also been utilized as a vaccine delivery platform for expression of diverse foreign antigens that stimulate protection against shigellosis, anthrax, plague, or human papilloma virus. However, Ty21a is acid-labile and, for effective oral immunization, stomach acidity has to be either neutralized with buffer or by-passed with Ty21a in an enteric-coated capsule (ECC). Several studies have shown that efficacy is reduced when Ty21a is administered in an ECC versus as a buffered liquid formulation, the former limiting exposure to GI tract lymphoid tissues. However, the ECC was selected as a more practical delivery format for both packaging/shipping and vaccine administration ease. We have sought to increase Ty21a acid-resistance to allow for removal from the ECC and immune enhancement. To improve Ty21a acid-resistance, glutamate-dependent acid resistance genes (GAD; responsible for Shigella spp. survival at very low pH) were cloned on a multi-copy plasmid (pGad) under a controllable arabinose-inducible promoter. pGad enhanced acid survival of Ty21a by 5 logs after 3 hours at pH 2.5, when cells were pre-grown in arabinose and under conditions that promote an acid-tolerance response (ATR). For genetically 100% stable expression, we inserted the gad genes into the Ty21a chromosome, using a method that allowed for subsequent removal of a selectable antibiotic resistance marker. Further, both bacterial growth curves and survival assays in cultured human monocytes/macrophages suggest that neither the genetic methods employed nor the resulting acid-resistance conferred by expression of the Gad proteins in Ty21a had any effect on the existing attenuation of this vaccine strain.

  9. Salmonella typhi vaccine strain CVD 908 expressing the circumsporozoite protein of Plasmodium falciparum: strain construction and safety and immunogenicity in humans.

    Science.gov (United States)

    Gonzalez, C; Hone, D; Noriega, F R; Tacket, C O; Davis, J R; Losonsky, G; Nataro, J P; Hoffman, S; Malik, A; Nardin, E; Sztein, M B; Heppner, D G; Fouts, T R; Isibasi, A; Levine, M M

    1994-04-01

    rcsp, encoding amino acids 21-398 of Plasmodium falciparum circumsporozoite protein (CSP), under control of tacP was integrated into the chromosomal delta aroC locus of attenuated delta aroC, delta aroD Salmonella typhi CVD 908. By immunoblot and ELISA, rCSP expression was greater from a multicopy plasmid than from the single chromosomal gene. CVD 908 omega (delta aroC1019::tacP-rcsp) was well tolerated by 10 volunteers who were fed two doses of 5 x 10(7) organisms 8 days apart. Seven subjects excreted the vaccine strain for 1-3 days. All subjects developed serologic responses to O and H antigens of the live vector, whereas 3 vaccinees responded to the foreign antigen: 1 developed an 80-fold rise in serum anti-sporozoite antibody, another had a 4-fold rise in antibody to a recombinant portion of CSP (residues 309-345), while a third vaccinee developed CSP-specific CD8+ cytotoxic T lymphocyte activity. This is the first report of attenuated S. typhi eliciting a human serologic or a cytotoxic T lymphocyte response to a foreign protein. Improved foreign gene expression should enhance immunogenicity.

  10. Antimicrobial susceptibility pattern and sequence analysis of DNA gyrase and DNA topoisomerase IV in Salmonella enterica serovars Typhi and Paratyphi A isolates with decreased susceptibility to ciprofloxacin.

    Science.gov (United States)

    Misra, Richa; Thakare, Ritesh; Amrin, N; Prasad, Kashi Nath; Chopra, Sidharth; Dhole, Tapan Nirodhechand

    2016-08-01

    We describe the antimicrobial susceptibility pattern of 100 typhoidal Salmonella isolates recovered from blood cultures and also investigate the association of decreased ciprofloxacin susceptibility with mutations in the genes coding for DNA gyrase and topoisomerase IV in 55 isolates. The study was conducted between January 2013 and December 2015 at a tertiary care centre in north India. Antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion and E-test. Genotypic characterization included the screening of mutations in the quinolone resistance-determining region of gyrA, gyrB, parC, and parE by PCR. DNA sequence analysis was done for 55 isolates. Out of 100 isolates recovered 80 were S. Typhi, 18 were Paratyphi A and two were Paratyphi B. Eighty two percent (66/80) of S. Typhi and 15/18 S. Paratyphi A showed decreased ciprofloxacin susceptibility. The most common mutation in gyrA led to a change at codon 83 of serine to phenylalanine (n=37) or tyrosine (n=12). Five S. Typhi isolates that were resistant to ciprofloxacin (MICs of 12, 16, 24 and 32 μg/ml) had a second mutation at codon 87 in the gyrA gene changing aspartate to asparagine. There is a need to urgently review the use of fluoroquinolones for the management of enteric fever in endemic areas. © The Author 2016. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Molecular structure of alpha-D-glucose-1-phosphate cytidylyltransferase from Salmonella typhi.

    Science.gov (United States)

    Koropatkin, Nicole M; Holden, Hazel M

    2004-10-15

    Dideoxysugars, which display biological activities ranging from mediating cell-cell interactions to serving as components in some antibiotics, are synthesized in various organisms via complex biochemical pathways that begin with the attachment of alpha-D-glucose 1-phosphate to either CTP or dTTP. Here we describe the three-dimensional structure of the alpha-D-glucose-1-phosphate cytidylyltransferase from Salmonella typhi, which catalyzes the first step in the production of CDP-tyvelose. For this investigation, the enzyme was crystallized in the presence of its product, CDP-glucose. In contrast to previous reports, the enzyme exists as a fully integrated hexamer with 32-point group symmetry. Each subunit displays a "bird-like" appearance with the "body" composed predominantly of a seven-stranded mixed beta-sheet and the two "wings" formed by beta-hairpin motifs. These two wings mediate subunit-subunit interactions along the 3-fold and 2-fold rotational axes, respectively. The six active sites of the hexamer are situated between the subunits related by the 2-fold rotational axes. CDP-glucose is anchored to the protein primarily by hydrogen bonds with backbone carbonyl oxygens and peptidic NH groups. The side chains of Arg111 and Asn188 from one subunit and Glu178 and Lys179 from the second subunit are also involved in hydrogen bonding with the ligand. The topology of the main core domain bears striking similarity to that observed for glucose-1-phosphate thymidylyltransferase and 4-diphosphocytidyl-2-C-methylerythritol synthetase.

  12. [Elaboration of an immunosorbent for the purification of porins from Salmonella typhi 9, 12, Vi:d].

    Science.gov (United States)

    Pelayo, R; Isibasi, A; Paniagua, J; Ortíz, V; Muy, M; González, C; Blanco, F; Islas, S; Kumate, J

    1989-01-01

    The current work was undertaken to purify porins of Salmonella typhi, which are outer membrane proteins (OMPs) that induce protection in mice against challenge with the bacteria in mucin. OMPs, isolated with a non-ionic detergent, had a 4% contamination with LPS (endotoxin) and molecular sizes ranging from 17 to 70 KDa. Porins (Mw 38-41 KDa) were isolated from OMPs preparative SDS-PAGE. Anti-porins antisera were raised in rabbits and specific IgG was purified, which was coupled to Sepharose-CNBr. This immunosorbent was used to purify LPS-free porins.

  13. Aktivitas Ekstrak Daun Kelor terhadap Respons Imun Humoral pada Mencit yang Diinfeksi Salmonella typhi (ACTIVITY OF KELOR LEAF EXTRACT ON HUMORAL IMMUNE RESPONSE IN MICE POST SALMONELLA TYPHI INFECTION

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    Mohammad Hefni

    2013-12-01

    Full Text Available The aim of this research was to analyze the activity of  kelor (Moringa oleifera Lam leaf extract onhumoral immune response in mice infected with Salmonella typhi. Mice were divided into two groups : non-infected and infectedS. typhi groups. Each group was administered orally for 20 days with varied doses ofkelor leaf extract i.e. dose (0 mg/kg BW, dose 1 (14 mg/kg BW, dose 2 (42 mg /kg BW, and dose 3 (84 mg/kg BW.  Then all of the sample in infected groups were injected with 108 cells S. typhi. The humoralimmunity responses were determined by observing the number of lymphoid B cell (B220 and naive Thecell (CD4+CD62L+ by using software BD CellQuest Flowcytometry. The data were analysed using Two-Way ANOVA (P<0.05, with SPSS 16.0 for Windows.  The kelor leaf extract showed imunostimulatoryactivity by significantly improved the number of lymphocyte B cell (B220,  and naive Th Cell (CD4+CD62L+in mice infected with S. typhi.  The lower doses (dose of 14 mg/kg BW, and 42 mg/kg BW of kelor leafextracts was more effective than the highest dose (84 mg/kg BW. On the other  hand, the high dose showedimunosupresor activity on naive Teessor Th Cell.  However, immunosupressor activity on naïve Th cell wasobserved on the mice given the highest dose of extract.

  14. Salmonella enterica Serotype Napoli is the First Cause of Invasive Nontyphoidal Salmonellosis in Lombardy, Italy (2010-2014), and Belongs to Typhi Subclade.

    Science.gov (United States)

    Huedo, Pol; Gori, Maria; Zolin, Anna; Amato, Ettore; Ciceri, Giulia; Bossi, Anna; Pontello, Mirella

    2017-03-01

    Salmonella enterica serotype Napoli (S. Napoli) is currently emerging in Europe and particularly in Italy, where in 2014 it caused a large outbreak associated with elevated rates of bacteremia. However, no study has yet investigated its invasive ability and phylogenetic classification. Here, we show that between 2010 and 2014, S. Napoli was the first cause of invasive salmonellosis affecting 40 cases out of 687 (invasive index: 5.8%), which is significantly higher than the invasive index of all the other nontyphoidal serotypes (2.0%, p < 0.05). Genomic and phylogenetic analyses of an invasive isolate revealed that S. Napoli belongs to Typhi subclade in clade A, Paratyphi A being the most related serotype and carrying almost identical pattern of typhoid-associated genes. This work presents evidence of invasive capacity of S. Napoli and argues for reconsideration of its nontyphoidal category.

  15. CRISPR is an optimal target for the design of specific PCR assays for salmonella enterica serotypes Typhi and Paratyphi A.

    Directory of Open Access Journals (Sweden)

    Laetitia Fabre

    Full Text Available BACKGROUND: Serotype-specific PCR assays targeting Salmonella enterica serotypes Typhi and Paratyphi A, the causal agents of typhoid and paratyphoid fevers, are required to accelerate formal diagnosis and to overcome the lack of typing sera and, in some situations, the need for culture. However, the sensitivity and specificity of such assays must be demonstrated on large collections of strains representative of the targeted serotypes and all other bacterial populations producing similar clinical symptoms. METHODOLOGY: Using a new family of repeated DNA sequences, CRISPR (clustered regularly interspaced short palindromic repeats, as a serotype-specific target, we developed a conventional multiplex PCR assay for the detection and differentiation of serotypes Typhi and Paratyphi A from cultured isolates. We also developed EvaGreen-based real-time singleplex PCR assays with the same two sets of primers. PRINCIPAL FINDINGS: We achieved 100% sensitivity and specificity for each protocol after validation of the assays on 188 serotype Typhi and 74 serotype Paratyphi A strains from diverse genetic groups, geographic origins and time periods and on 70 strains of bacteria frequently encountered in bloodstream infections, including 29 other Salmonella serotypes and 42 strains from 38 other bacterial species. CONCLUSIONS: The performance and convenience of our serotype-specific PCR assays should facilitate the rapid and accurate identification of these two major serotypes in a large range of clinical and public health laboratories with access to PCR technology. These assays were developed for use with DNA from cultured isolates, but with modifications to the assay, the CRISPR targets could be used in the development of assays for use with clinical and other samples.

  16. [Sensitivity to chlorine-containing disinfectants of Salmonella typhi strains with transmissible drug resistance].

    Science.gov (United States)

    Karmanova, G I; Morozova, N S; Emel'ianova, O I; Kibardina, N N

    1987-02-01

    Sensitivity of 201 S. typhi strains to antibiotics and chlorine containing disinfectants was studied. The strains were isolated in the regions of the Left Bank Ukraine. Among the typhoid cultures the proportion of R+ strains was on the whole equal to 29.3 per cent. Chloramine resistant strains amounted to 21.8 per cent. Among chloramine resistant strains the number of the cultures carrying R plasmids was 3 times higher than that of the strains sensitive to disinfectant. Simultaneous transfer of the R plasmid and chloramine resistance to the recipient strain suggested that resistance of the S. typhi strains to chloramine was of plasmid nature.

  17. Differential epidemiology of Salmonella Typhi and Paratyphi A in Kathmandu, Nepal: a matched case control investigation in a highly endemic enteric fever setting.

    Science.gov (United States)

    Karkey, Abhilasha; Thompson, Corinne N; Tran Vu Thieu, Nga; Dongol, Sabina; Le Thi Phuong, Tu; Voong Vinh, Phat; Arjyal, Amit; Martin, Laura B; Rondini, Simona; Farrar, Jeremy J; Dolecek, Christiane; Basnyat, Buddha; Baker, Stephen

    2013-01-01

    Enteric fever, a systemic infection caused by the bacteria Salmonella Typhi and Salmonella Paratyphi A, is endemic in Kathmandu, Nepal. Previous work identified proximity to poor quality water sources as a community-level risk for infection. Here, we sought to examine individual-level risk factors related to hygiene and sanitation to improve our understanding of the epidemiology of enteric fever in this setting. A matched case-control analysis was performed through enrollment of 103 blood culture positive enteric fever patients and 294 afebrile community-based age and gender-matched controls. A detailed questionnaire was administered to both cases and controls and the association between enteric fever infection and potential exposures were examined through conditional logistic regression. Several behavioral practices were identified as protective against infection with enteric fever, including water storage and hygienic habits. Additionally, we found that exposures related to poor water and socioeconomic status are more influential in the risk of infection with S. Typhi, whereas food consumption habits and migration play more of a role in risk of S. Paratyphi A infection. Our work suggests that S. Typhi and S. Paratyphi A follow different routes of infection in this highly endemic setting and that sustained exposure to both serovars probably leads to the development of passive immunity. In the absence of a polyvalent vaccine against S. Typhi and S. Paratyphi A, we advocate better systems for water treatment and storage, improvements in the quality of street food, and vaccination with currently available S. Typhi vaccines.

  18. Níveis de sensibilidade de enterobactérias, em particular Salmonella typhi, a rifamicina S.V.

    Directory of Open Access Journals (Sweden)

    Ivone R. Suassuna

    1969-04-01

    Full Text Available Em face à predominante eliminação biliar da rifamicina S.V. atingindo concentrações muitas vêzes superiores aos níveis séricos obtidos com as doses terapêuticas, e pelo possível interêsse dessa verificação para o tratamento dos portadores biliares crônicos de Salmonella typhi determinou-se a concentração mínima inibitória de 165 estirpes de enterobactérias, incluindo 77 amostras de S. typhi. Foi verificado que a maioria das cepas de Escherichia coli, Shigella e Proteus mirabilis correspondiam a uma concentração inibitória mínima entre 33 a 65 μg/ml. Entre 65 e 128 μg/ml foram determinadas as concetrações inibitórias mínimas da maioria das outras espécies de Proteus, de Providencia e de Klebsiella. Para Salmonella e Enterobacter o limite mínino de sensibilidade foi, em regra, igual ou superior a 128 μg/ml. Diferenças mais acentuadas de comportamento entre as enterobactérias foram observadas quanto à ação bactericida da rifamicidas S.V. De uma maneira geral, para E. coli e Shigella, as concentrações inibitórias mínimas já referidas. Para as espécies de Proteus e Providencia houve variação maior de comportamento, mas tendência a que o efeito bactericidas fôsse encontrado em concentrações que correspondiam a 4 vêzes as bacteriostáticas para as mesmas espécies. Finalmente, de modo pouco feliz para os propósitos visados, em Salmonella, com a inclusão de S. typhi, não foi atingido um efeito bactericida, com as mais altas concentrações usadas as quais corresponderam em média a 6 vêzes as concentrações bacteriostáticas para esse gênero.

  19. Oral Challenge with Wild-Type Salmonella Typhi Induces Distinct Changes in B Cell Subsets in Individuals Who Develop Typhoid Disease.

    Directory of Open Access Journals (Sweden)

    Franklin R Toapanta

    2016-06-01

    Full Text Available A novel human oral challenge model with wild-type Salmonella Typhi (S. Typhi was recently established by the Oxford Vaccine Group. In this model, 104 CFU of Salmonella resulted in 65% of participants developing typhoid fever (referred here as typhoid diagnosis -TD- 6-9 days post-challenge. TD was diagnosed in participants meeting clinical (oral temperature ≥38°C for ≥12h and/or microbiological (S. Typhi bacteremia endpoints. Changes in B cell subpopulations following S. Typhi challenge remain undefined. To address this issue, a subset of volunteers (6 TD and 4 who did not develop TD -NoTD- was evaluated. Notable changes included reduction in the frequency of B cells (cells/ml of TD volunteers during disease days and increase in plasmablasts (PB during the recovery phase (>day 14. Additionally, a portion of PB of TD volunteers showed a significant increase in activation (CD40, CD21 and gut homing (integrin α4β7 molecules. Furthermore, all BM subsets of TD volunteers showed changes induced by S. Typhi infections such as a decrease in CD21 in switched memory (Sm CD27+ and Sm CD27- cells as well as upregulation of CD40 in unswitched memory (Um and Naïve cells. Furthermore, changes in the signaling profile of some BM subsets were identified after S. Typhi-LPS stimulation around time of disease. Notably, naïve cells of TD (compared to NoTD volunteers showed a higher percentage of cells phosphorylating Akt suggesting enhanced survival of these cells. Interestingly, most these changes were temporally associated with disease onset. This is the first study to describe differences in B cell subsets directly related to clinical outcome following oral challenge with wild-type S. Typhi in humans.

  20. Oral Challenge with Wild-Type Salmonella Typhi Induces Distinct Changes in B Cell Subsets in Individuals Who Develop Typhoid Disease.

    Science.gov (United States)

    Toapanta, Franklin R; Bernal, Paula J; Fresnay, Stephanie; Magder, Laurence S; Darton, Thomas C; Jones, Claire; Waddington, Claire S; Blohmke, Christoph J; Angus, Brian; Levine, Myron M; Pollard, Andrew J; Sztein, Marcelo B

    2016-06-01

    A novel human oral challenge model with wild-type Salmonella Typhi (S. Typhi) was recently established by the Oxford Vaccine Group. In this model, 104 CFU of Salmonella resulted in 65% of participants developing typhoid fever (referred here as typhoid diagnosis -TD-) 6-9 days post-challenge. TD was diagnosed in participants meeting clinical (oral temperature ≥38°C for ≥12h) and/or microbiological (S. Typhi bacteremia) endpoints. Changes in B cell subpopulations following S. Typhi challenge remain undefined. To address this issue, a subset of volunteers (6 TD and 4 who did not develop TD -NoTD-) was evaluated. Notable changes included reduction in the frequency of B cells (cells/ml) of TD volunteers during disease days and increase in plasmablasts (PB) during the recovery phase (>day 14). Additionally, a portion of PB of TD volunteers showed a significant increase in activation (CD40, CD21) and gut homing (integrin α4β7) molecules. Furthermore, all BM subsets of TD volunteers showed changes induced by S. Typhi infections such as a decrease in CD21 in switched memory (Sm) CD27+ and Sm CD27- cells as well as upregulation of CD40 in unswitched memory (Um) and Naïve cells. Furthermore, changes in the signaling profile of some BM subsets were identified after S. Typhi-LPS stimulation around time of disease. Notably, naïve cells of TD (compared to NoTD) volunteers showed a higher percentage of cells phosphorylating Akt suggesting enhanced survival of these cells. Interestingly, most these changes were temporally associated with disease onset. This is the first study to describe differences in B cell subsets directly related to clinical outcome following oral challenge with wild-type S. Typhi in humans.

  1. Oral Wild-Type Salmonella Typhi Challenge Induces Activation of Circulating Monocytes and Dendritic Cells in Individuals Who Develop Typhoid Disease.

    Science.gov (United States)

    Toapanta, Franklin R; Bernal, Paula J; Fresnay, Stephanie; Darton, Thomas C; Jones, Claire; Waddington, Claire S; Blohmke, Christoph J; Dougan, Gordon; Angus, Brian; Levine, Myron M; Pollard, Andrew J; Sztein, Marcelo B

    2015-06-01

    A new human oral challenge model with wild-type Salmonella Typhi (S. Typhi) was recently developed. In this model, ingestion of 104 CFU of Salmonella resulted in 65% of subjects developing typhoid fever (referred here as typhoid diagnosis -TD-) 5-10 days post-challenge. TD criteria included meeting clinical (oral temperature ≥38°C for ≥12 h) and/or microbiological (S. Typhi bacteremia) endpoints. One of the first lines of defense against pathogens are the cells of the innate immune system (e.g., monocytes, dendritic cells -DCs-). Various changes in circulating monocytes and DCs have been described in the murine S. Typhimurium model; however, whether similar changes are present in humans remains to be explored. To address these questions, a subset of volunteers (5 TD and 3 who did not develop typhoid despite oral challenge -NoTD-) were evaluated for changes in circulating monocytes and DCs. Expression of CD38 and CD40 were upregulated in monocytes and DCs in TD volunteers during the disease days (TD-0h to TD-96h). Moreover, integrin α4β7, a gut homing molecule, was upregulated on monocytes but not DCs. CD21 upregulation was only identified in DCs. These changes were not observed among NoTD volunteers despite the same oral challenge. Moreover, monocytes and DCs from NoTD volunteers showed increased binding to S. Typhi one day after challenge. These monocytes showed phosphorylation of p38MAPK, NFkB and Erk1/2 upon stimulation with S. Typhi-LPS-QDot micelles. In contrast, monocytes from TD volunteers showed only a moderate increase in S. Typhi binding 48 h and 96 h post-TD, and only Erk1/2 phosphorylation. This is the first study to describe different activation and migration profiles, as well as differential signaling patterns, in monocytes and DCs which relate directly to the clinical outcome following oral challenge with wild type S. Typhi.

  2. The influence of MAP condition and active compounds on the radiosensitization of Escherichia coli and Salmonella typhi present in chicken breast

    Energy Technology Data Exchange (ETDEWEB)

    Lacroix, M. E-mail: monique.lacroix@inrs-iaf.uquebec.ca; Chiasson, F

    2004-10-01

    The efficiency of carvacrol, thymol, trans-cinnamaldehyde (Tc) and tetrasodium pyrophosphate (Tp) on the radiosensitization of Escherichia coli and Salmonella typhi in chicken breast was determined. Chicken breast were dipped in a bath of working cultures of E. coli or S. typhi (5x10{sup 7} CFU/ml). Active compounds were added at the concentration corresponding to ((1)/(30)) of the minimal inhibitory concentration. Samples were packed under air and gamma irradiation was done at doses from 0.1 to 0.7 kGy. The efficiencies of the active compounds against E. coli were 32%, 10%, 3% and 0% for thymol, Tp and carvacrol, respectively. For S. typhi, the efficiencies in the chicken breast were 47%, 19%, 17% and 11% for Tc, Tp, carvacrol and thymol, respectively. Without active compounds, D{sub 10} values were 0.145 kGy for E. coli and 0.64 kGy for S. typhi as compared to 0.098 kGy for E. coli and 0.341 kGy for S. typhi in presence of Tc. Under modified atmospheric packaging condition and in presence of Tc, D{sub 10} values were reduced to 0.046 for E. coli and to 0.110 for S. typhi.

  3. [About problems of carriers of Salmonella typhi and paratyphi B under hygienic, epidemiologic and sociologic view (authors transl)].

    Science.gov (United States)

    Brossmann, D

    1977-01-01

    The carriers of salmonella typhi and paratyphi B are the central figures of the epidemiology of typhoid illnesses. The files available at the medical authorities' office of the Hansestadt Lübeck, with regards to all 543 carriers registered up to 1972, have been analysed, and 102 of 108 carriers who were inhabitants of Lübeck on 31.10.72, have been interviewed in their lodgings.--Following results are mentionable: 1. 55 carriers have been traced during a hospital stay, 22 of them were under hospital treatment because of cholecystectomy. 2. The circle of persons to be obligated for medical examination as per $ 17 of the Federal Republic of Germany's law of epidemics has proved as too small. For instance two female carriers--known since years--did their job without any hindering as plates washer or waitress in restaurants. 3. 21 persons=4% of the patients excreted salmonellae in urine only. 4. 10% of the carriers of salmonella typhi and 16% of the carriers of salmonella paratypi B had in their faeces as least 5-years intervals of non-excretion. 5. The precentage of carriers with at least 5-years intervals of non-excretion in urine was significantly larger than the corresponding percentage in faeces. 6. Falling ill of 280 persons out of the environs of the carriers have been lead back to the latters. 73% of these infections were placed by the carriers before their detection through the medical authorities' office. 7. For 10 patients whose health was recreated excretion did not cease promptly after the cholecystecystectomy. Salmonellae had been found in the faeces for several months more, at two persons even two years after the operation. 8. For 8 carriers a final decision with regards to the success of cholecystectomy is impossible because they denied a duodenal soundation. 9. 123 persons of 23% of total 543 carriers had--before their ascertation--professions not allowed for those persons. Efforts for a successful job arrangement, possibly combined with a change in

  4. Stochastic simulation of endemic Salmonella enterica serovar Typhi: the importance of long lasting immunity and the carrier state.

    Directory of Open Access Journals (Sweden)

    Allan Saul

    Full Text Available BACKGROUND: Typhoid fever caused by Salmonella enterica serovar Typhi (S. Typhi remains a serious burden of disease, especially in developing countries of Asia and Africa. It is estimated that it causes 200,000 deaths per year, mainly in children. S. Typhi is an obligate pathogen of humans and although it has a relatively complex life cycle with a long lived carrier state, the absence of non-human hosts suggests that well targeted control methods should have a major impact on disease. Newer control methods including new generations of vaccines offer hope but their implementation would benefit from quantitative models to guide the most cost effective strategies. This paper presents a quantitative model of Typhoid disease, immunity and transmission as a first step in that process. METHODOLOGY/PRINCIPAL FINDINGS: A stochastic agent-based model has been developed that incorporates known features of the biology of typhoid including probability of infection, the consequences of infection, treatment options, acquisition and loss of immunity as a result of infection and vaccination, the development of the carrier state and the impact of environmental or behavioral factors on transmission. The model has been parameterized with values derived where possible from the literature and where this was not possible, feasible parameters space has been determined by sensitivity analyses, fitting the simulations to age distribution of field data. The model is able to adequately predict the age distribution of typhoid in two settings. CONCLUSIONS/SIGNIFICANCE: The modeling highlights the importance of variations in the exposure/resistance of infants and young children to infection in different settings, especially as this impacts on design of control programs; it predicts that naturally induced clinical and sterile immunity to typhoid is long lived and highlights the importance of the carrier state especially in areas of low transmission.

  5. The effects of Brazilian and Bulgarian propolis in vitro against Salmonella Typhi and their synergism with antibiotics acting on the ribosome.

    Science.gov (United States)

    Orsi, R O; Fernandes, A; Bankova, V; Sforcin, J M

    2012-01-01

    Salmonella enterica serovar Typhi is the causative agent of typhoid fever in humans, and the use of antibiotics is essential for controlling this infection; however, the excessive use of antibiotics may select resistant strains. Propolis is a honeybee product and its antimicrobial activity has been intensively investigated. Thus, the objective of this study was to investigate a possible synergism between propolis (collected in Brazil and Bulgaria) and antibiotics acting on the ribosome (chloramphenicol, tetracycline and neomycin) against Salmonella Typhi in vitro. The synergism was investigated by using ½ and ¼ of the minimum inhibitory concentration for propolis and these antimicrobial agents, evaluating the number of viable cells according to the incubation time. Brazilian propolis showed a bacteriostatic action against S. Typhi, while Bulgarian propolis showed a bactericidal activity and a synergistic effect with the three antibiotics. Variations in the biological assays might be due to the differences in their chemical compositions. Based on the results, one may conclude that Bulgarian propolis showed an important antibacterial action, as well as a synergistic effect with antibiotics acting on the ribosome, which points out a possible therapeutic strategy evaluating the use of propolis preparations for the treatment of Salmonella Typhi infection.

  6. PRODUCTION OF BACTERIOCIN EC2 AND ITS INTERFERENCE IN THE GROWTH OF SALMONELLA TYPHI IN A MILK MATRIX

    Directory of Open Access Journals (Sweden)

    Yuri de Jesus Lopes de Abreu

    2013-08-01

    Full Text Available Bacterial interference can occur through various mechanisms, including the production of peroxides, acids, ammonia, bacteriolytic enzymes or bacteriocins. The strain Escherichia coli EC2 produces the antimicrobial substance (AMS EC2, able to inhibit different strains of Gram-negative bacteria isolated from food, as E. coli and Salmonella sp. The activity of AMS EC2 was lost after treatment with proteolytic enzymes, indicating the presence of an active proteinaceous compound, suggesting that it is a bacteriocin. The substance, renamed bacteriocin EC2, has its better production when the producer strain is grown on Casoy medium, at 37ºC and pH 6.0, without NaCl addition, but it is also able to be produced in milk. When co-cultivated in UHT milk with the producer strain E. coli EC2, the growth of the indicator strain Salmonella Typhi is totally inhibited within the first 4 hours of incubation, suggesting a potential application of bacteriocin EC2 in the control of Salmonella sp. e.g. in foods.

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

    Directory of Open Access Journals (Sweden)

    Angela M Jansen

    2011-07-01

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

  8. Genomic Dissection of Travel-Associated Extended-Spectrum-Beta-Lactamase-Producing Salmonella enterica Serovar Typhi Isolates Originating from the Philippines: a One-Off Occurrence or a Threat to Effective Treatment of Typhoid Fever?

    DEFF Research Database (Denmark)

    Hendriksen, Rene S.; Leekitcharoenphon, Pimlapas; Mikoleit, Matthew

    2015-01-01

    One unreported case of extended-spectrum-beta-lactamase (ESBL)-producing Salmonella enterica serovar Typhi was identified, whole-genome sequence typed, among other analyses, and compared to other available genomes of S. Typhi. The reported strain was similar to a previously published strain harbo...

  9. Phylogeographical analysis of the dominant multidrug-resistant H58 clade of Salmonella Typhi identifies inter- and intracontinental transmission events

    Science.gov (United States)

    Wong, Vanessa K; Baker, Stephen; Pickard, Derek J; Parkhill, Julian; Page, Andrew J; Feasey, Nicholas A; Kingsley, Robert A; Thomson, Nicholas R; Keane, Jacqueline A; Weill, François-Xavier; Edwards, David J; Hawkey, Jane; Harris, Simon R; Mather, Alison E; Cain, Amy K; Hadfield, James; Hart, Peter J; Thieu, Nga Tran Vu; Klemm, Elizabeth J; Glinos, Dafni A; Breiman, Robert F; Watson, Conall H; Kariuki, Samuel; Gordon, Melita A; Heyderman, Robert S; Okoro, Chinyere; Jacobs, Jan; Lunguya, Octavie; Edmunds, W John; Msefula, Chisomo; Chabalgoity, Jose A; Kama, Mike; Jenkins, Kylie; Dutta, Shanta; Marks, Florian; Campos, Josefina; Thompson, Corinne; Obaro, Stephen; MacLennan, Calman A; Dolecek, Christiane; Keddy, Karen H; Smith, Anthony M; Parry, Christopher M; Karkey, Abhilasha; Mulholland, E Kim; Campbell, James I; Dongol, Sabina; Basnyat, Buddha; Dufour, Muriel; Bandaranayake, Don; Naseri, Take Toleafoa; Singh, Shalini Pravin; Hatta, Mochammad; Newton, Paul; Onsare, Robert S; Isaia, Lupeoletalalei; Dance, David; Davong, Viengmon; Thwaites, Guy; Wijedoru, Lalith; Crump, John A; De Pinna, Elizabeth; Nair, Satheesh; Nilles, Eric J; Thanh, Duy Pham; Turner, Paul; Soeng, Sona; Valcanis, Mary; Powling, Joan; Dimovski, Karolina; Hogg, Geoff; Farrar, Jeremy; Holt, Kathryn E; Dougan, Gordon

    2016-01-01

    The emergence of multidrug-resistant (MDR) typhoid is a major global health threat affecting many countries where the disease is endemic. Here whole-genome sequence analysis of 1,832 Salmonella enterica serovar Typhi (S. Typhi) identifies a single dominant MDR lineage, H58, that has emerged and spread throughout Asia and Africa over the last 30 years. Our analysis identifies numerous transmissions of H58, including multiple transfers from Asia to Africa and an ongoing, unrecognized MDR epidemic within Africa itself. Notably, our analysis indicates that H58 lineages are displacing antibiotic-sensitive isolates, transforming the global population structure of this pathogen. H58 isolates can harbor a complex MDR element residing either on transmissible IncHI1 plasmids or within multiple chromosomal integration sites. We also identify new mutations that define the H58 lineage. This phylogeographical analysis provides a framework to facilitate global management of MDR typhoid and is applicable to similar MDR lineages emerging in other bacterial species. PMID:25961941

  10. Structural basis of typhoid: Salmonella typhi type IVb pilin (PiLS) and cystic fibrosis transmembrane conductance regulator interaction

    Energy Technology Data Exchange (ETDEWEB)

    Balakrishna, A.M.; Saxena, A.; Mok, H. Y.-K.; Swaminathan, K.

    2009-11-01

    The type IVb pilus of the enteropathogenic bacteria Salmonella typhi is a major adhesion factor during the entry of this pathogen into gastrointestinal epithelial cells. Its target of adhesion is a stretch of 10 residues from the first extracellular domain of cystic fibrosis transmembrane conductance regulator (CFTR). The crystal structure of the N-terminal 25 amino acid deleted S. typhi native PilS protein ({Delta}PilS), which makes the pilus, was determined at 1.9 {angstrom} resolution by the multiwavelength anomalous dispersion method. Also, the structure of the complex of {Delta}PilS and a target CFTR peptide, determined at 1.8 {angstrom}, confirms that residues 113-117 (NKEER) of CFTR are involved in binding with the pilin protein and gives us insight on the amino acids that are essential for binding. Furthermore, we have also explored the role of a conserved disulfide bridge in pilus formation. The subunit structure and assembly architecture are crucial for understanding pilus functions and designing suitable therapeutics against typhoid.

  11. Vaccination with Salmonella Typhi recombinant outer membrane protein 28 induces humoral but non-protective immune response in rabbit

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    Anjani Saxena

    2017-08-01

    Full Text Available Aim: Typhoid is one of the most important food and water borne disease causing millions of deaths over the world. Presently, there is no cost effective vaccine available in India. The outer-membrane proteins (Omps of Salmonella have been exhibited as a potential candidate for development of subunit vaccine against typhoid. The objective of the present study was to evaluate the use of recombinant Omp 28 protein for immunization of rabbit to elucidate its protection against virulent Salmonella Typhi. Materials and Methods: Immune potential of recombinant Omp28 was tested in New Zealand Rabbits. Rabbits were divided into two groups, i.e., control and test group. Control group was injected with phosphate buffer saline with adjuvant while test group were injected with recombinant Omp28 along with adjuvant. Rabbits were bleed and serum was collected from each rabbit. Serum was tested by Enzyme-linked immunosorbent assay (ELISA for humoral response. Rabbits were challenged with virulent culture to test the protective immunity. Results: Humoral response was provoked at 15th day and maintained till 30th day. The mean ELISA titer at 15th day was 1 : 28000 (mean titer log 10 : 4.4472 and on the 30th day was 1 : 25866 (mean titer log 10 : 4.4127. Protective immune potential of Omp 28 was assessed by challenge studies in rabbits for which vaccinated and control rabbits were challenged with 109 cells of virulent culture of S. Typhi. In control group, out of six, no rabbit could survive after 48 days while in vaccinated group, three out of six rabbit were survived. Conclusion: Immunization of rabbit with recombinant Omp 28 induced a strong humoral response which was exhibited by high antibody titer in ELISA. Subsequently, intraperitoneal homologous challenge of the immunized New Zealand rabbit resulted in lack of significant protection. These findings indicate that Omp 28 though provoked the humoral immunity but could not provide the protective immunity in

  12. Characterization of putative multidrug resistance transporters of the major facilitator-superfamily expressed in Salmonella Typhi

    DEFF Research Database (Denmark)

    Shaheen, Aqsa; Ismat, Fouzia; Iqbal, Mazhar

    2015-01-01

    Multidrug resistance mediated by efflux pumps is a well-known phenomenon in infectious bacteria. Although much work has been carried out to characterize multidrug efflux pumps in Gram-negative and Gram-positive bacteria, such information is still lacking for many deadly pathogens. The aim...... conferred resistance to at least ten of the tested antimicrobials: ciprofloxacin, norfloxacin, levofloxacin, kanamycin, streptomycin, gentamycin, nalidixic acid, chloramphenicol, ethidium bromide, and acriflavine, including fluoroquinolone antibiotics, which were drugs of choice to treat S. Typhi infections...

  13. Challenge of Humans with Wild-type Salmonella enterica Serovar Typhi Elicits Changes in the Activation and Homing Characteristics of Mucosal-Associated Invariant T Cells.

    Science.gov (United States)

    Salerno-Goncalves, Rosângela; Luo, David; Fresnay, Stephanie; Magder, Laurence; Darton, Thomas C; Jones, Claire; Waddington, Claire S; Blohmke, Christoph J; Angus, Brian; Levine, Myron M; Pollard, Andrew J; Sztein, Marcelo B

    2017-01-01

    Gastrointestinal infections by Salmonella enterica serovar Typhi (S. Typhi) are rare in industrialized countries. However, they remain a major public health problem in the developing world with an estimated 26.9 million new cases annually and significant mortality when untreated. Recently, we provided the first direct evidence that CD8+ MAIT cells are activated and have the potential to kill cells exposed to S. Typhi, and that these responses are dependent on bacterial load. However, MAIT cell kinetics and function during bacterial infections in humans remain largely unknown. In this study, we characterize the human CD8+ MAIT cell immune response to S. Typhi infection in subjects participating in a challenge clinical trial who received a low- or high dose of wild-type S. Typhi. We define the kinetics of CD8+ MAIT cells as well as their levels of activation, proliferation, exhaustion/apoptosis, and homing potential. Regardless of the dose, in volunteers resistant to infection (NoTD), the levels of CD8+ MAIT cells after S. Typhi challenge fluctuated around their baseline values (day 0). In contrast, volunteers susceptible to the development of typhoid disease (TD) exhibited a sharp decline in circulating MAIT cells during the development of typhoid fever. Interestingly, MAIT cells from low-dose TD volunteers had higher levels of CD38 coexpressing CCR9, CCR6, and Ki67 during the development of typhoid fever than high-dose TD volunteers. No substantial perturbations on the levels of these markers were observed in NoTD volunteers irrespective of the dose. In sum, we describe, for the first time, that exposure to an enteric bacterium, in this case S. Typhi, results in changes in MAIT cell activation, proliferation, and homing characteristics, suggesting that MAIT cells are an important component of the human host response to bacterial infection.

  14. Evolution of Salmonella Typhi outer membrane protein-specific T and B cell responses in humans following oral Ty21a vaccination: A randomized clinical trial.

    Directory of Open Access Journals (Sweden)

    Juan Manuel Carreño

    Full Text Available Vaccination against complex pathogens such as typhoidal and non-typhoidal Salmonella requires the concerted action of different immune effector mechanisms. Outer membrane proteins (Omps of Salmonella Typhi are potent immunogens, which elicit long-lasting and protective immunity. Here, we followed the evolution of S. Typhi OmpC and F-specific T and B cell responses in healthy volunteers after vaccination with the vaccine strain Ty21a. To follow humoral and cellular immune responses, pre- and post-vaccination samples (PBMC, serum and stool collected from 15 vaccinated and 5 non-vaccinated individuals. Immunoglobulin levels were assessed in peripheral blood by enzyme-linked immunosorbent assay. B cell and T cell activation were analyzed by flow cytometry. We observed a significant increase of circulating antibody-secreting cells and maximal Omp-specific serum IgG titers at day 25 post vaccination, while IgA titers in stool peaked at day 60. Likewise, Omp-specific CD4+ T cells in peripheral blood showed the highest expansion at day 60 post vaccination, concomitant with a significant increase in IFN-γ and TNFα production. These results indicate that S. Typhi Omp-specific B cell responses and polyfunctional CD4+ T cell responses evolve over a period of at least two months after application of the live attenuated vaccine. Moreover, these findings underscore the potential of S. Typhi Omps as subunit vaccine components.ISRCTN18360696.

  15. Immunoblot detection of class-specific humoral immune response to outer membrane proteins isolated from Salmonella typhi in humans with typhoid fever.

    OpenAIRE

    ORTIZ,V; Isibasi, A.; García-Ortigoza, E; Kumate, J.

    1989-01-01

    The studies reported here were undertaken to assess the ability of the outer membrane proteins (OMPs) of Salmonella typhi to induce a humoral immune response in humans with typhoid fever. OMPs were isolated with the nonionic detergent Triton X-100 and were found to be contaminated with approximately 4% lipopolysaccharide. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns showed protein bands with molecular size ranges from 17 to 70 kilodaltons; the major groups of proteins we...

  16. Salmonella typhi: lisotipia VI e biotipificação em amostras oriundas de algumas regiões do Brasil Salmonella typhi: lysotype VI and biotyping in sample from some regions of Brazil

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    Ernesto Hofer

    1984-03-01

    Full Text Available Fez-se uma análise da distribuicão da frequência dos lisotipos VI e dos tipos fermentativos segundo o esquema de Kristensen, em 1.150 amostras de Salmonella typhi, isoladas de diferentes regiões do Brasil (Pará, Pernambuco, Bahia, Minas Gerais, Rio de Janeiro, São Paulo e Rio Grande do Sul. No computo geral, observou-se a prevalência dos lisotipos A (38,1%; Ela (18,9%; amostras VI negativas (16,6%; D6 (8,7% I + IV (4,6%; T (2,3% e C1 (2,1% e a ocorrência de alguns tipos fágicos característicos para determinadas áreas (B3, C4 e 40 na Bahia; E1b, F2, G1 e L1 em São Paulo; E4 e 28 no Rio de Janeiro. Quanto a classificacão bioquímica, 55,2% das amostras caracterizaram-se no biotipo II (xilose e arabinose negativas, 44,2% no tipo fermentativo I (xilose positiva e arabinose negativas e 0,52% no tipo III (xilose e arabinose positivas, respectivamente.The frequency of Vi-phage types and fermentative types according to Kristensen's scheme was studied among 1,150 strains of Salmonella typhi isolated from different areas in Brazil (states of Pará, Pernambuco, Bahia, Minas Gerais, Rio de Janeiro, São Paulo and Rio Grande do Sul. The most prevalent phage types encountered in this study were: A (38.1%; Ela (18.9%, D6 (8.7%, T (2.3% and C1 (2.1%, including categories of untypable strains (group I + IV-4.6%, and Vi negative (16.6%. There was,however, some types characteristics of particular areas (B3, C4, 40 from Bahia; Elb, F2,G1, L1 from São Paulo; E4 and 28 from Rio de Janeiro. In respect to the biochemical classification, 55.2% of the strains were classified as a biotype II (xylose and arabinose negative, 44,2% as of type I (xylose positive and negative0 and 0.52% as a type III (xylose and arabinose positive, respectively.

  17. [Epidemiological characteristics of typhoid fever and antibiotic susceptibility testing of Salmonella Typhi isolates in Guangxi, 1994-2013].

    Science.gov (United States)

    Wang, Mingliu; Kan, Biao; Yang, Jin; Lin, Mei; Yan, Meiying; Zeng, Jun; Quan, Yi; Liao, Hezhuang; Zhou, Lingyun; Jiang, Zhenling; Huang, Dehui

    2014-08-01

    Through analyzing the typhoid epidemics and to determine and monitor regional resistance characteristics of the shift of drug resistant profile on Salmonella (S.) Typhi, to understand the related epidemiological characteristics of typhoid fever and to provide evidence for the development of strategies, in Guangxi. Data of typhoid fever from surveillance and reporting system between 1994 to 2013 was collected and statistically analyzed epidemiologically. The susceptibility of 475 S. Typhi isolates from patients on ten antibiotics was tested by broth micro-dilution method and minimum inhibition concentration was obtained and interpreted based on the CLSI standard. From 1994 to 2013, a total of 57 928 cases of typhoid fever were reported in Guangxi province with an annual incidence of 6.29/100 000 and mortality as 0.03%. The higher incidence was observed in the population under 20 years of age. There was no significant difference on incidence between male and female, but farmers and students were among the hardest hit groups. More cases were seen from the northern part of the province. Cases appeared all year round with the peak from May to October. A total of 13 major outbreaks during 2001 to 2013 were reported and the main transmission route was water-borne. All the strains were sensitive to third generation cephalosporins cefotaxime and fluoroquinolones norfloxacin. The susceptibility rates to tetracycline, chloramphenicol, ampicillin and gentamicin was around 98% but relative lower susceptible rate to ciprofloxacin was seen as 89.89% . The lowest susceptibility was found for streptomycin and sulfamethoxazole agents, with the rates as 67.73% and 65.89% , respectively. One strain was found to have been resistant to ciprofloxacin and another 47 isolates with reduced susceptibility to ciprofloxacin. Twenty eight isolates were found to be resistant to multiple antibiotics and one displayed ampicillin, chloramphenicol, streptomycin, sulfamethoxazole tetracycline and

  18. Differential epidemiology of Salmonella Typhi and Paratyphi A in Kathmandu, Nepal: a matched case control investigation in a highly endemic enteric fever setting.

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    Abhilasha Karkey

    Full Text Available Enteric fever, a systemic infection caused by the bacteria Salmonella Typhi and Salmonella Paratyphi A, is endemic in Kathmandu, Nepal. Previous work identified proximity to poor quality water sources as a community-level risk for infection. Here, we sought to examine individual-level risk factors related to hygiene and sanitation to improve our understanding of the epidemiology of enteric fever in this setting.A matched case-control analysis was performed through enrollment of 103 blood culture positive enteric fever patients and 294 afebrile community-based age and gender-matched controls. A detailed questionnaire was administered to both cases and controls and the association between enteric fever infection and potential exposures were examined through conditional logistic regression. Several behavioral practices were identified as protective against infection with enteric fever, including water storage and hygienic habits. Additionally, we found that exposures related to poor water and socioeconomic status are more influential in the risk of infection with S. Typhi, whereas food consumption habits and migration play more of a role in risk of S. Paratyphi A infection.Our work suggests that S. Typhi and S. Paratyphi A follow different routes of infection in this highly endemic setting and that sustained exposure to both serovars probably leads to the development of passive immunity. In the absence of a polyvalent vaccine against S. Typhi and S. Paratyphi A, we advocate better systems for water treatment and storage, improvements in the quality of street food, and vaccination with currently available S. Typhi vaccines.

  19. Emergence of plasmid-mediated multidrug resistance in epidemic and non-epidemic strains of Salmonella enterica serotype Typhi from Jordan.

    Science.gov (United States)

    Al-Sanouri, Tarek M; Paglietti, Bianca; Haddadin, Aktham; Murgia, Manuela; Bacciu, Donatella; Youssef, Mahammad; Rubino, Salvatore

    2008-08-30

    Enteric fever caused by Salmonella enterica serovar Typhi has not been adequately explored in Jordan. In this study we investigated antibiotic resistance patterns and resistance determinants coupled with fingerprint methods of forty-eight isolates of S. Typhi obtained from 113 patients with suspected enteric fever admitted at six governmental hospitals in different directorates in Jordan. Twenty-four isolates were from an outbreak of typhoid fever that occurred between October 2004 and January 2005, and another twenty-four were from sporadic cases from 2005. All isolates of S. Typhi were resistant to streptomycin. A multidrug resistant (MDR) pattern of ampicillin, chloramphenicol, co-trimoxazole with tetracycline and streptomycin (R-type ACCoTS) was found in 58% of the epidemic strains causing the outbreak and in 98% of the strains from sporadic cases. MDR isolates harbored a single IncHI1 plasmid containing a class 1 integron (dfrA7). Plasmid conjugation studies demonstrated a genetic transfer of resistance (ACCoT). S. Typhi isolates were all sensitive to fluoroquinolones and cefotaxime, the alternative drugs recommended for treatment of typhoid fever. The genomic analysis using PFGE showed: a) the outbreak was caused by an introduced circulating clone with/without an MDR plasmid, and b) isolates from the sporadic cases from 2005 are the same MDR clone that persisted and spread in the country. The emergence of MDR S. Typhi strains is a majorn important public health issue in Jordan. This study should guide selection of effective antibiotic therapy for the treatment of typhoid and monitoring of the spread of MDR of S. Typhi.

  20. Selection of autochthonous lactic acid bacteria from goat dairies and their addition to evaluate the inhibition of Salmonella typhi in artisanal cheese.

    Science.gov (United States)

    Ferrari, Iris da Silva; de Souza, Jane Viana; Ramos, Cintia Lacerda; da Costa, Mateus Matiuzzi; Schwan, Rosane Freitas; Dias, Francesca Silva

    2016-12-01

    This study aimed to select autochthonous lactic acid bacteria (LAB) with probiotic and functional properties from goat dairies and test their addition to artisanal cheese for the inhibition of Salmonella typhi. In vitro tests, including survival in the gastrointestinal tract (GIT), auto- and co-aggregation, the hemolytic test, DNase activity, antimicrobial susceptibility, antibacterial activity, tolerance to NaCl and exopolysaccharide (EPS), gas and diacetyl production were conducted for sixty isolates. Based on these tests, four LAB isolates (UNIVASF CAP 16, 45, 84 and 279) were selected and identified. Additional tests, such as production of lactic and citric acids by UNIVASF CAP isolates were performed in addition to assays of bile salt hydrolase (BSH), β-galactosidase and decarboxylase activity. The four selected LAB produced high lactic acid (>17 g/L) and low citric acid (0.2 g/L) concentrations. All selected strains showed BSH and β-galactosidase activity and none showed decarboxylase activity. Three goat cheeses (1, 2 and control) were produced and evaluated for the inhibitory action of selected LAB against Salmonella typhi. The cheese inoculated with LAB (cheese 2) decreased 0.38 log10 CFU/g of S. Typhy population while in the cheese without LAB inoculation (cheese 1) the pathogen population increased by 0.29 log units. Further, the pH value increased linearly over time, by 0.004 units per day in cheese 1. In the cheese 2, the pH value decreased linearly over time, by 0.066 units per day. The cocktail containing selected Lactobacillus strains with potential probiotic and technological properties showed antibacterial activity against S. typhi in vitro and in artisanal goat cheese. Thus, goat milk is important source of potential probiotic LAB which may be used to inhibit the growth of Salmonella population in cheese goat, contributing to safety and functional value of the product. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Oral Wild-Type Salmonella Typhi Challenge Induces Activation of Circulating Monocytes and Dendritic Cells in Individuals Who Develop Typhoid Disease.

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    Franklin R Toapanta

    2015-06-01

    Full Text Available A new human oral challenge model with wild-type Salmonella Typhi (S. Typhi was recently developed. In this model, ingestion of 104 CFU of Salmonella resulted in 65% of subjects developing typhoid fever (referred here as typhoid diagnosis -TD- 5-10 days post-challenge. TD criteria included meeting clinical (oral temperature ≥38°C for ≥12 h and/or microbiological (S. Typhi bacteremia endpoints. One of the first lines of defense against pathogens are the cells of the innate immune system (e.g., monocytes, dendritic cells -DCs-. Various changes in circulating monocytes and DCs have been described in the murine S. Typhimurium model; however, whether similar changes are present in humans remains to be explored. To address these questions, a subset of volunteers (5 TD and 3 who did not develop typhoid despite oral challenge -NoTD- were evaluated for changes in circulating monocytes and DCs. Expression of CD38 and CD40 were upregulated in monocytes and DCs in TD volunteers during the disease days (TD-0h to TD-96h. Moreover, integrin α4β7, a gut homing molecule, was upregulated on monocytes but not DCs. CD21 upregulation was only identified in DCs. These changes were not observed among NoTD volunteers despite the same oral challenge. Moreover, monocytes and DCs from NoTD volunteers showed increased binding to S. Typhi one day after challenge. These monocytes showed phosphorylation of p38MAPK, NFkB and Erk1/2 upon stimulation with S. Typhi-LPS-QDot micelles. In contrast, monocytes from TD volunteers showed only a moderate increase in S. Typhi binding 48 h and 96 h post-TD, and only Erk1/2 phosphorylation. This is the first study to describe different activation and migration profiles, as well as differential signaling patterns, in monocytes and DCs which relate directly to the clinical outcome following oral challenge with wild type S. Typhi.

  2. Genotypic characterization of Salmonella typhi by amplified fragment length polymorphism fingerprinting provides increased discrimination as compared to pulsed-field gel electrophoresis and ribotyping.

    Science.gov (United States)

    Nair, S; Schreiber, E; Thong, K L; Pang, T; Altwegg, M

    2000-06-01

    Amplified fragment length polymorphism (AFLP) is a recently developed, PCR-based high resolution fingerprinting method that is able to generate complex banding patterns which can be used to delineate intraspecific genetic relationships among bacteria. In the present study, AFLP was evaluated for its usefulness in the molecular typing of Salmonella typhi in comparison to ribotyping and pulsed-field gel electrophoresis (PFGE). Six S. typhi isolates from diverse geographic areas (Malaysia, Indonesia, India, Chile, Papua New Guinea and Switzerland) gave unique, heterogeneous profiles when typed by AFLP, a result which was consistent with ribotyping and PFGE analysis. In a further study of selected S. typhi isolates from Papua New Guinea which caused fatal and non-fatal disease previously shown to be clonally related by PFGE, AFLP discriminated between these isolates but did not indicate a linkage between genotype with virulence. We conclude that AFLP (discriminatory index=0.88) has a higher discriminatory power for strain differentiation among S. typhi than ribotyping (DI=0.63) and PFGE (DI=0.74).

  3. Synergistic antimicrobial effect of nisin and p-cymene on Salmonella enterica serovar Typhi in vitro and on ready-to-eat food.

    Science.gov (United States)

    Rattanachaikunsopon, Pongsak; Phumkhachorn, Parichat

    2010-01-01

    Foods contaminated with Salmonella enterica serovar Typhi are a major cause of typhoid fever, leading to public health problems and economic losses worldwide. Nisin and rho-cymene were tested in this study for their antimicrobial activity against S. Typhi at 4 degrees C and 37 degrees C. Nisin and rho-cymene, when used separately, did not inhibit the bacterium at either temperature. A synergistic antimicrobial effect between both compounds was observed when they were used simultaneously. This synergism was greater at 37 degrees C than at 4 degrees C. The lowest concentrations of nisin and rho-cymene required for complete inhibition of S. Typhi at 37 degrees C were 0.3 ppm and 1.5 ppm, respectively, and 0.3 ppm and 2.5 ppm at 4 degrees C. The potential of nisin and rho-cymene to control an S. Typhi population on ready-to-eat Thai-style pork sausage was also examined. The compounds were able to eliminate the contaminating bacterium with concentrations depending on the bacterial cell number on the food.

  4. Validation and application of an ELISA for the quantification of IgG antibodies against Salmonella Typhi Vi capsular polysaccharide

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    Yanet Martin-García

    2015-08-01

    Full Text Available An indirect ELISA for the quantification of IgG antibodies against the Vi polysaccharide of this bacteria was developed as a part of the stages of Research and Development of a conjugate vaccinal candidate against Salmonella Typhi. The results of the validation process are presented in this paper, in which the interval and linearity of the curve, the intra- and inter-assay precision, accuracy, specificity, limit of detection and robustness were determined. The calibration curve generated with an internal standard serum provided a good fit to a polynomial function and an interval between 1/100 and 1/3200 dilutions. The coefficients of variation in the precision and robustness tests and the percentages of recovered were in intervals established for each one (≤10%, ≤20% and 90-110%, respectively. The assay presented an optimal specificity, obtaining OD signals above 1.3 for positive sera against Vi and low for sera against unrelated antigens. The results support the use of this quantitative ELISA in immunogenicity assays for batch release of Vi conjugates. Likewise, they support their use for the immunogenicity evaluation of Vi polysaccharide formulations and Vi polysaccharide conjugates to proteins in phases of research and development

  5. ANTIBACTERIAL EFFECT OF GARLIC (ALLIUM SATIVUM AND GINGER (ZINGIBER OFFICINALE AGAINST STAPHYLOCOCCUS AUREUS, SALMONELLA TYPHI, ESCHERICHIA COLI AND BACILLUS CEREUS

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    Bandna Chand

    2013-02-01

    Full Text Available Antibacterial activity of extracts of Allium sativum (garlic and Zingiber officinale (ginger has been evaluated against four different bacteria namely Escherichia coli, Salmonella Typhi, Staphylococcus aureus and Bacillus cereus. Two methods were used to determine the antimicrobial activity of garlic and ginger extracts namely disk diffusion method and agar well diffusion method. Garlic extract exhibited excellent antibacterial activity against all four test organisms while ginger extract showed antibacterial activity against Bacillus cereus and Staphylococcus aureus only. In addition, agar well diffusion method showed higher zone in inhibition when compared with the zone of inhibition produced by the spice of same concentration against the test microorganism by disk diffusion method. Antibiotic sensitivity of the four different bacteria was tested with commercially available antibiotics namely Ciprofloxacin; Oxytetracycline; Vancomycin; Streptomycin; Gentamicin; Tetracycline; Novobiocin; Amikacin and Penicillin G. Penicillin G produced the highest zone of inhibition of 40.00±0.00against Staphylococcus aureus and the lowest zone of inhibition of 0.00±0.00against Escherichia coli.

  6. Investigations of binding mode insight in Salmonella typhi type-III secretion system tip protein (SipD: A molecular docking and MD simulation study

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    Gopinath Samykannu

    2017-01-01

    Full Text Available Salmonella typhi is a Gram-negative pathogen that utilizes type three secretion systems (TTSS to translocate virulence factors into host cells. This causes a wide variety of diseases mainly gastroenteritis and typhoid fever in both humans and animals. Salmonella invasive protein D (SipD is a tip protein, attached to the needle protein and interacts with translocon complex. Exploring the molecular mechanisms of SipD may aid in better understanding about secretion, translocators and transmission of environmental signals of Salmonella. In this study, SipD sequence (Q56136 was retrieved from Uniprot for which three-dimensional (3D structure was modelled with the help of I-TASSER server. The modelled structure was subjected to molecular docking using Glide. About 25 compounds of T3SS inhibitors were chosen after literature studies and docking was performed. It was evident that polyol product Caminoside A possessed (≤-5 kcal/mol the best interaction with SipD and therefore this complex was subjected to molecular dynamic simulation for interaction stability analysis. The simulation results showed that Caminoside A binding was highly stable (life time of 80% with SipD and Asn318 interaction was more significant for the protein-ligand complex stability. These results will provide new knowledge for the development of novel therapeutic strategies against Salmonella typhi.

  7. Fifth Percentile Cutoff Values for Antipneumococcal Polysaccharide and Anti-Salmonella typhi Vi IgG Describe a Normal Polysaccharide Response

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    Heidi Schaballie

    2017-05-01

    Full Text Available BackgroundSerotype-specific antibody responses to unconjugated pneumococcal polysaccharide vaccine (PPV evaluated by a World Health Organization (WHO-standardized enzyme-linked immunosorbent assay (ELISA are the gold standard for diagnosis of specific polysaccharide antibody deficiency (SAD. The American Academy of Allergy, Asthma and Immunology (AAAAI has proposed guidelines to interpret the PPV response measured by ELISA, but these are based on limited evidence. Additionally, ELISA is costly and labor-intensive. Measurement of antibody response to Salmonella typhi (S. typhi Vi vaccine and serum allohemagglutinins (AHA have been suggested as alternatives. However, there are no large cohort studies and cutoff values are lacking.ObjectiveTo establish cutoff values for antipneumococcal polysaccharide antibody response, anti-S. typhi Vi antibody, and AHA.MethodsOne hundred healthy subjects (10–55 years were vaccinated with PPV and S. typhi Vi vaccine. Blood samples were obtained prior to and 3–4 weeks after vaccination. Polysaccharide responses to 3 serotypes were measured by WHO ELISA and to 12 serotypes by an in-house bead-based multiplex assay. Anti-S. typhi Vi IgG were measured with a commercial ELISA kit. AHA were measured by agglutination method.ResultsApplying AAAAI criteria, 30% of healthy subjects had a SAD. Using serotype-specific fifth percentile (p5 cutoff values for postvaccination IgG and fold increase pre- over postvaccination, only 4% of subjects had SAD. One-sided 95% prediction intervals for anti-S. typhi Vi postvaccination IgG (≥11.2 U/ml and fold increase (≥2 were established. Eight percent had a response to S. typhi Vi vaccine below these cutoffs. AHA titer p5 cutoffs were ½ for anti-B and ¼ for anti-A.ConclusionWe establish reference cutoff values for interpretation of PPV response measured by bead-based assay, cutoff values for S. typhi Vi vaccine responses, and normal values for AHA. For the first time, the

  8. Fifth Percentile Cutoff Values for Antipneumococcal Polysaccharide and Anti-Salmonella typhi Vi IgG Describe a Normal Polysaccharide Response.

    Science.gov (United States)

    Schaballie, Heidi; Bosch, Barbara; Schrijvers, Rik; Proesmans, Marijke; De Boeck, Kris; Boon, Mieke Nelly; Vermeulen, François; Lorent, Natalie; Dillaerts, Doreen; Frans, Glynis; Moens, Leen; Derdelinckx, Inge; Peetermans, Willy; Kantsø, Bjørn; Jørgensen, Charlotte Svaerke; Emonds, Marie-Paule; Bossuyt, Xavier; Meyts, Isabelle

    2017-01-01

    Serotype-specific antibody responses to unconjugated pneumococcal polysaccharide vaccine (PPV) evaluated by a World Health Organization (WHO)-standardized enzyme-linked immunosorbent assay (ELISA) are the gold standard for diagnosis of specific polysaccharide antibody deficiency (SAD). The American Academy of Allergy, Asthma and Immunology (AAAAI) has proposed guidelines to interpret the PPV response measured by ELISA, but these are based on limited evidence. Additionally, ELISA is costly and labor-intensive. Measurement of antibody response to Salmonella typhi (S. typhi) Vi vaccine and serum allohemagglutinins (AHA) have been suggested as alternatives. However, there are no large cohort studies and cutoff values are lacking. To establish cutoff values for antipneumococcal polysaccharide antibody response, anti-S. typhi Vi antibody, and AHA. One hundred healthy subjects (10-55 years) were vaccinated with PPV and S. typhi Vi vaccine. Blood samples were obtained prior to and 3-4 weeks after vaccination. Polysaccharide responses to 3 serotypes were measured by WHO ELISA and to 12 serotypes by an in-house bead-based multiplex assay. Anti-S. typhi Vi IgG were measured with a commercial ELISA kit. AHA were measured by agglutination method. Applying AAAAI criteria, 30% of healthy subjects had a SAD. Using serotype-specific fifth percentile (p5) cutoff values for postvaccination IgG and fold increase pre- over postvaccination, only 4% of subjects had SAD. One-sided 95% prediction intervals for anti-S. typhi Vi postvaccination IgG (≥11.2 U/ml) and fold increase (≥2) were established. Eight percent had a response to S. typhi Vi vaccine below these cutoffs. AHA titer p5 cutoffs were ½ for anti-B and ¼ for anti-A. We establish reference cutoff values for interpretation of PPV response measured by bead-based assay, cutoff values for S. typhi Vi vaccine responses, and normal values for AHA. For the first time, the intraindividual consistency of all three methods is

  9. Optimization of randomly amplified polymorphic DNA-polymerase chain reaction for molecular typing of Salmonella enterica serovar Typhi Otimização da reação de amplificação aleatória do DNA polimórfico - reação em cadeia da polimerase para tipagem molecular de Salmonella enterica sorovar Typhi

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    Bianca Ramalho Quintaes

    2004-03-01

    Full Text Available Optimization of the RAPD reaction for characterizing Salmonella enterica serovar Typhi strains was studied in order to ensure the reproducibility and the discriminatory power of this technique. Eight Salmonella serovar Typhi strains isolated from various regions in Brazil were examined for the fragment patterns produced using different concentrations of DNA template, primer, MgCl2 and Taq DNA polymerase. Using two different low stringency thermal cycle profiles, the RAPD fingerprints obtained were compared. A set of sixteen primers was evaluated for their ability to produce a high number of distinct fragments. We found that variations associated to all of the tested parameters modified the fingerprinting patterns. For the strains of Salmonella enterica serovar Typhi used in this experiment, we have defined a set of conditions for RAPD-PCR reaction, which result in a simple, fast and reproducible typing method.A otimização da reação de RAPD para a caracterização de cepas de Salmonella enterica sorovar Typhi foi estudada com o objetivo de assegurar a reprodutibilidade e o poder discriminatório desta técnica. Oito cepas de Salmonella sorovar Typhi isoladas de algumas regiões do Brasil foram usadas para examinar os padrões de fragmentação produzidos quando foram empregadas concentrações diferentes do DNA molde, do iniciador, do MgCl2 e da enzima Taq DNA polimerase. Com a utilização de dois diferentes perfis de ciclos termais de baixa estringência, foram comparados os padrões de bandeamento obtidos. Um conjunto de dezesseis iniciadores foi avaliado quanto à capacidade de produzir elevado número de fragmentos distintos. Observou-se que variações associadas a todos os parâmetros testados modificaram os padrões de bandeamento. Para as amostras de Salmonella enterica sorovar Typhi utilizadas neste experimento, definiu-se um conjunto de condições para a reação de RAPD-PCR que resultou num método de tipagem simples, rápido e

  10. An outpatient, ambulant-design, controlled human infection model using escalating doses of Salmonella Typhi challenge delivered in sodium bicarbonate solution.

    Science.gov (United States)

    Waddington, Claire S; Darton, Thomas C; Jones, Claire; Haworth, Kathryn; Peters, Anna; John, Tessa; Thompson, Ben A V; Kerridge, Simon A; Kingsley, Robert A; Zhou, Liqing; Holt, Kathryn E; Yu, Ly-Mee; Lockhart, Stephen; Farrar, Jeremy J; Sztein, Marcelo B; Dougan, Gordon; Angus, Brian; Levine, Myron M; Pollard, Andrew J

    2014-05-01

    Typhoid fever is a major global health problem, the control of which is hindered by lack of a suitable animal model in which to study Salmonella Typhi infection. Until 1974, a human challenge model advanced understanding of typhoid and was used in vaccine development. We set out to establish a new human challenge model and ascertain the S. Typhi (Quailes strain) inoculum required for an attack rate of 60%-75% in typhoid-naive volunteers when ingested with sodium bicarbonate solution. Groups of healthy consenting adults ingested escalating dose levels of S. Typhi and were closely monitored in an outpatient setting for 2 weeks. Antibiotic treatment was initiated if typhoid diagnosis occurred (temperature ≥38°C sustained ≥12 hours or bacteremia) or at day 14 in those remaining untreated. Two dose levels (10(3) or 10(4) colony-forming units) were required to achieve the primary objective, resulting in attack rates of 55% (11/20) or 65% (13/20), respectively. Challenge was well tolerated; 4 of 40 participants fulfilled prespecified criteria for severe infection. Most diagnoses (87.5%) were confirmed by blood culture, and asymptomatic bacteremia and stool shedding of S. Typhi was also observed. Participants who developed typhoid infection demonstrated serological responses to flagellin and lipopolysaccharide antigens by day 14; however, no anti-Vi antibody responses were detected. Human challenge with a small inoculum of virulent S. Typhi administered in bicarbonate solution can be performed safely using an ambulant-model design to advance understanding of host-pathogen interactions and immunity. This model should expedite development of diagnostics, vaccines, and therapeutics for typhoid control.

  11. An Outpatient, Ambulant-Design, Controlled Human Infection Model Using Escalating Doses of Salmonella Typhi Challenge Delivered in Sodium Bicarbonate Solution

    Science.gov (United States)

    Waddington, Claire S.; Darton, Thomas C.; Jones, Claire; Haworth, Kathryn; Peters, Anna; John, Tessa; Thompson, Ben A. V.; Kerridge, Simon A.; Kingsley, Robert A.; Zhou, Liqing; Holt, Kathryn E.; Yu, Ly-Mee; Lockhart, Stephen; Farrar, Jeremy J.; Sztein, Marcelo B.; Dougan, Gordon; Angus, Brian; Levine, Myron M.; Pollard, Andrew J.

    2014-01-01

    Background. Typhoid fever is a major global health problem, the control of which is hindered by lack of a suitable animal model in which to study Salmonella Typhi infection. Until 1974, a human challenge model advanced understanding of typhoid and was used in vaccine development. We set out to establish a new human challenge model and ascertain the S. Typhi (Quailes strain) inoculum required for an attack rate of 60%–75% in typhoid-naive volunteers when ingested with sodium bicarbonate solution. Methods. Groups of healthy consenting adults ingested escalating dose levels of S. Typhi and were closely monitored in an outpatient setting for 2 weeks. Antibiotic treatment was initiated if typhoid diagnosis occurred (temperature ≥38°C sustained ≥12 hours or bacteremia) or at day 14 in those remaining untreated. Results. Two dose levels (103 or 104 colony-forming units) were required to achieve the primary objective, resulting in attack rates of 55% (11/20) or 65% (13/20), respectively. Challenge was well tolerated; 4 of 40 participants fulfilled prespecified criteria for severe infection. Most diagnoses (87.5%) were confirmed by blood culture, and asymptomatic bacteremia and stool shedding of S. Typhi was also observed. Participants who developed typhoid infection demonstrated serological responses to flagellin and lipopolysaccharide antigens by day 14; however, no anti-Vi antibody responses were detected. Conclusions. Human challenge with a small inoculum of virulent S. Typhi administered in bicarbonate solution can be performed safely using an ambulant-model design to advance understanding of host–pathogen interactions and immunity. This model should expedite development of diagnostics, vaccines, and therapeutics for typhoid control. PMID:24519873

  12. Slide Coagglutination for Salmonella typhi Antigens in Broths Inoculated with Feces from Typhoid Fever Patients

    Science.gov (United States)

    1981-12-01

    hoeae, N. meningitidis and Lancefield groups sis of typhoid fever were history of fever for A, B, D and G streptococci (Danielsson and at least seven...SUKSANoNG, M. and DAJANT, A.S., (1977). 144. -a .. . ’U 532 Vol. 12 No. 4 December 1981 U U UNCLASSIFIED SECURITY CLASSIFICATION OF THIS PAGE (When...Twelve of these later seroconverted to Salmonella 0 -j DO A’In 1473 UNCLASSIFIED .over SECURITY CLASSIFICATION OF THIS P-G--(W fte - .Entered.) F

  13. Differing Burden and Epidemiology of Non-Typhi Salmonella Bacteremia in Rural and Urban Kenya, 2006–2009

    Science.gov (United States)

    Tabu, Collins; Breiman, Robert F.; Ochieng, Benjamin; Aura, Barrack; Cosmas, Leonard; Audi, Allan; Olack, Beatrice; Bigogo, Godfrey; Ongus, Juliette R.; Fields, Patricia; Mintz, Eric; Burton, Deron; Oundo, Joe; Feikin, Daniel R.

    2012-01-01

    Background The epidemiology of non-Typhi Salmonella (NTS) bacteremia in Africa will likely evolve as potential co-factors, such as HIV, malaria, and urbanization, also change. Methods As part of population-based surveillance among 55,000 persons in malaria-endemic, rural and malaria-nonendemic, urban Kenya from 2006–2009, blood cultures were obtained from patients presenting to referral clinics with fever ≥38.0°C or severe acute respiratory infection. Incidence rates were adjusted based on persons with compatible illnesses, but whose blood was not cultured. Results NTS accounted for 60/155 (39%) of blood culture isolates in the rural and 7/230 (3%) in the urban sites. The adjusted incidence in the rural site was 568/100,000 person-years, and the urban site was 51/100,000 person-years. In both sites, the incidence was highest in children 85% of blood NTS isolates in both sites, but only 21% (urban) and 64% (rural) of stool NTS isolates. Overall, 76% of S. Typhimurium blood isolates were multi-drug resistant, most of which had an identical profile in Pulse Field Gel Electrophoresis. In the rural site, the incidence of NTS bacteremia increased during the study period, concomitant with rising malaria prevalence (monthly correlation of malaria positive blood smears and NTS bacteremia cases, Spearman's correlation, p = 0.018 for children, p = 0.16 adults). In the rural site, 80% of adults with NTS bacteremia were HIV-infected. Six of 7 deaths within 90 days of NTS bacteremia had HIV/AIDS as the primary cause of death assigned on verbal autopsy. Conclusions NTS caused the majority of bacteremias in rural Kenya, but typhoid predominated in urban Kenya, which most likely reflects differences in malaria endemicity. Control measures for malaria, as well as HIV, will likely decrease the burden of NTS bacteremia in Africa. PMID:22363591

  14. Differing burden and epidemiology of non-Typhi Salmonella bacteremia in rural and urban Kenya, 2006-2009.

    Directory of Open Access Journals (Sweden)

    Collins Tabu

    Full Text Available BACKGROUND: The epidemiology of non-Typhi Salmonella (NTS bacteremia in Africa will likely evolve as potential co-factors, such as HIV, malaria, and urbanization, also change. METHODS: As part of population-based surveillance among 55,000 persons in malaria-endemic, rural and malaria-nonendemic, urban Kenya from 2006-2009, blood cultures were obtained from patients presenting to referral clinics with fever ≥38.0°C or severe acute respiratory infection. Incidence rates were adjusted based on persons with compatible illnesses, but whose blood was not cultured. RESULTS: NTS accounted for 60/155 (39% of blood culture isolates in the rural and 7/230 (3% in the urban sites. The adjusted incidence in the rural site was 568/100,000 person-years, and the urban site was 51/100,000 person-years. In both sites, the incidence was highest in children 85% of blood NTS isolates in both sites, but only 21% (urban and 64% (rural of stool NTS isolates. Overall, 76% of S. Typhimurium blood isolates were multi-drug resistant, most of which had an identical profile in Pulse Field Gel Electrophoresis. In the rural site, the incidence of NTS bacteremia increased during the study period, concomitant with rising malaria prevalence (monthly correlation of malaria positive blood smears and NTS bacteremia cases, Spearman's correlation, p = 0.018 for children, p = 0.16 adults. In the rural site, 80% of adults with NTS bacteremia were HIV-infected. Six of 7 deaths within 90 days of NTS bacteremia had HIV/AIDS as the primary cause of death assigned on verbal autopsy. CONCLUSIONS: NTS caused the majority of bacteremias in rural Kenya, but typhoid predominated in urban Kenya, which most likely reflects differences in malaria endemicity. Control measures for malaria, as well as HIV, will likely decrease the burden of NTS bacteremia in Africa.

  15. PARTICIPACION DE LOS MARCOS DE LECTURA STY1362, STY1364 y STY1365 DE SALMONELLA ENTERICA SEROVAR TYPHI EN LA INTERACCION CON EL MACROFAGO HUMANO Y LA ENVOLTURA BACTERIANA

    OpenAIRE

    RODAS GARRIDO, PAULA

    2012-01-01

    Salmonella enterica incluye a un amplio grupo de microorganismos Gram-negativos y patógenos intracelulares facultativos que infectan un amplio rango de hospederos y causan una variedad de enfermedades que van desde una gastroenteritis limitada hasta una severa infección sistémica. De ellos, Salmonella enterica serovar Typhi (S. Typhi) infecta exclusivamente al ser humano y ha demostrado ser un patógeno altamente adaptado a su hospedero, siendo el agente etiológico de la fiebre tifoidea...

  16. DNA biosensor for detection of Salmonella typhi from blood sample of typhoid fever patient using gold electrode modified by self-assembled monolayers of thiols

    Science.gov (United States)

    Suryapratiwi, Windha Novita; Paat, Vlagia Indira; Gaffar, Shabarni; Hartati, Yeni Wahyuni

    2017-05-01

    Electrochemical biosensors are currently being developed in order to handle various clinical problems in diagnosing infectious diseases caused by pathogenic bacteria, or viruses. On this research, voltammetric DNA biosensor using gold electrode modified by thiols with self-assembled monolayers had been developed to detect a certain sequence of Salmonella typhi DNA from blood sample of typhoid fever patient. Thiol groups of cysteamines (Cys) and aldehyde groups from glutaraldehydes (Glu) were used as a link to increase the performance of gold electrode in detecting guanine oxidation signal of hybridized S. typhi DNA and ssDNA probe. Standard calibration method was used to determine analytical parameters from the measurements. The result shown that, the detection of S. typhi DNA from blood sample of typhoid fever patient can be carried out by voltammetry using gold electrode modified by self-assembled monolayers of thiols. A characteristic oxidation potential of guanine using Au/Cys/Gluwas obtained at +0.17 until +0.20 V. Limit of detection and limit of quantification from this measurements were 1.91μg mL-1 and 6.35 μg mL-1. The concentration of complement DNA from sample was 6.96 μg mL-1.

  17. Characterization of Citrobacter sp. line 328 as a source of Vi for a Vi-CRM(197) glycoconjugate vaccine against Salmonella Typhi.

    Science.gov (United States)

    Rondini, Simona; Micoli, Francesca; Lanzilao, Luisa; Pisoni, Ivan; Di Cioccio, Vito; Saul, Allan J; Martin, Laura B

    2012-11-26

    Salmonella enterica serovar Typhi is the causative agent of typhoid fever with over 22 million cases and over 200,000 deaths reported annually. A vaccine is much needed for use in young children and the Novartis Vaccines Institute for Global Health (NVGH) is developing a conjugate vaccine which targets S. Typhi Vi capsular polysaccharide. Here we describe a method suitable for industrial scale production of the Vi antigen based on expression by a Citrobacter line. We optimized the production of Vi by selecting a suitable Citrobacter strain (Citrobacter 328) that yields high and stable expression of Vi in chemically defined medium under industrial-scale fermentation conditions. Vi-CRM197 made using Vi from Citrobacter 328 elicited high anti-Vi antibody levels in mice and rabbits. Citrobacter 328 is a suitable strain for production of Vi for conjugate anti-Typhi vaccines. Being a BSL-1 organism, which grows in defined medium and stably produces high yields of Vi, it offers excellent potential for safe production of inexpensive vaccines for populations at risk of typhoid fever.

  18. Immunoblot detection of class-specific humoral immune response to outer membrane proteins isolated from Salmonella typhi in humans with typhoid fever.

    Science.gov (United States)

    Ortiz, V; Isibasi, A; García-Ortigoza, E; Kumate, J

    1989-07-01

    The studies reported here were undertaken to assess the ability of the outer membrane proteins (OMPs) of Salmonella typhi to induce a humoral immune response in humans with typhoid fever. OMPs were isolated with the nonionic detergent Triton X-100 and were found to be contaminated with approximately 4% lipopolysaccharide. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns showed protein bands with molecular size ranges from 17 to 70 kilodaltons; the major groups of proteins were those that correspond to the porins and OmpA of gram-negative bacteria. Rabbit antiserum to OMPs or to S. typhi recognized OMPs after absorption with lipopolysaccharide. Sera from patients with typhoid fever contained immunoglobulin M antibodies which reacted with a protein of 28 kilodaltons and immunoglobulin G antibodies which reacted mainly with the porins, as determined by immunoblotting. These results indicate that the porins are the major immunogenic OMPs from S. typhi and that the immune response induced in the infection could be related to the protective status.

  19. Effect of selected plant essential oils or their constituents and modified atmosphere packaging on the radiosensitivity of Escherichia coli O157:H7 and Salmonella typhi in ground beef.

    Science.gov (United States)

    Turgis, M; Borsa, J; Millette, M; Salmieri, S; Lacroix, M

    2008-03-01

    Twenty-six different essential oils were tested for their efficiency to increase the relative radiosensitivity of Escherichia coli and Salmonella Typhi in medium-fat ground beef (23% fat). Ground beef was inoculated with E. coli O157:H7 or Salmonella (10(6) CFU/g), and each essential oil or one of their main constituents was added separately at a concentration of 0.5% (wt/wt). Meat samples (10 g) were packed under air or under modified atmosphere and irradiated at doses from 0 to 1 kGy for the determination of the D10-value of E. coli O157:H7, and from 0 to 1.75 kGy for the determination of the D10-value of Salmonella Typhi. Depending on the compound tested, the relative radiation sensitivity increased from 1 to 3.57 for E. coli O157:H7 and from 1 to 3.26 for Salmonella Typhi. Addition of essential oils or their constituents before irradiation also reduced the irradiation dose needed to eliminate both pathogens. In the presence of Chinese cinnamon or Spanish oregano essential oils, the minimum doses required to eliminate the bacteria were reduced from 1.2 to 0.35 and from 1.4 to 0.5 for E. coli O157:H7 and Salmonella Typhi, respectively. Cinnamon, oregano, and mustard essential oils were the most effective radiosensitizers.

  20. Validation of a traditional preparation against multi-drug resistant Salmonella Typhi and its protective efficacy in S. Typhimurium infected mice.

    Science.gov (United States)

    Chattopadhyay, Debprasad; Ojha, Durbadal; Mukherjee, Hemanta; Bag, Paromita; Vaidya, Shatrughna Prasad; Dutta, Shanta

    2018-01-17

    Shorea robusta Gaertn has been used for skin and intestinal ailments in Indian Traditional medicine; while two tribal communities used its tender leaves in 'Meyadi-bukhar' or long-term fever. This prompted us to validate the aqueous and methanol extracts of Shorea robusta tender leaves against wild- and multidrug-resistant clinical isolates of Salmonella enterica Serovar Typhi (S. Typhi), the causative agent of typhoid fever. The antibacterial activity, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and growth inhibition were determined using disc diffusion, agar-and-broth dilution, dose- and time-response assays, along with the safety and protective efficacy in Balb/C mice, infected with S. Typhimurium. The MIC of the extract was 256-450 μg/ml against S. Typhi isolates, and 700 μg/ml for mouse virulent S. Typhimurium, while MBC was ≤512-1024 μg/ml. The growth curve revealed that the extract was bactericidal at 4-6 h of exposure. Toxicity study showed that the extract was safe up to 3000 mg/kg (p.o.). Moreover, it significantly (p > 0.01) protect the challenged (1.4 × 10 8  cfu/ml) mice at 93.75 (i.p.) and 300 mg/kg (p.o.) dose, compared to the infection control (distilled water treatment group). Collectively, our results confirmed the antibacterial potential of the test extracts against MDR-isolates of S. Typhi. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  1. Evaluation of Dri-Dot OMPs Salmonella Typhi in Suspected Typhoid Fever Patients as an Immunodiagnostic Kit

    OpenAIRE

    Natzir, Rosdiana

    2016-01-01

    - Background. Several studies have identified a protein of OMP S. Typhi. The protein is highly immunogenic and can be recognize whole cells of S. Typhi. The aims of the study was to evaluate the Dri-dot OMPs immunoreactivity in typhoid fever suspected patients. Method. Samples obtained from Dr. Wahidin Sudirohusodo Hospital, lbnu Sina Hospital, and Haji Hospital of Makassar from February to November 2014. Their sensitivity and specificity were evaluated against blood culture method as the ...

  2. Preliminary Investigation of Bioactive Compounds and Bioautographic Studies of Whole Plant Extract of Euphorbia pulcherrima on Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    H. B. Sharif

    2015-01-01

    Full Text Available The aim of this study is to carry out preliminary investigation of bioactive compounds and bioautographic studies of whole plant extract of Euphorbia pulcherrima on Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Pseudomonas aeruginosa. Tukey HSD test of hierarchy for the effect of different solvents crude extract on bacterial isolates indicates the methanol extract as the most bioactive. The Tukey HSD analysis also showed that the bioactivities of the crude extracts of the various parts of Euphorbia pulcherrima were part dependent and the whole plant was the most bioactive. The ethyl acetate fraction of the methanol extract of the whole plant of Euphorbia pulcherrima has been shown in this work to contain phytochemicals which have shown remarkable activities against Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Pseudomonas aeruginosa. The bioactivities against the test organisms were due to the combined effects of the compounds separated on TLC plates. Families of terpenoids, flavonoids, alkaloids, saponin, and steroids that were detected in the extracts were identified by GC-MS. The various classes of phytochemicals in the E. pulcherrima plant provided the antimicrobial potency of the plant.

  3. Enhancement of ovarian cancer chemotherapy by delivery of multidrug-resistance gene small interfering RNA using tumor targeting Salmonella.

    Science.gov (United States)

    Deng, Jiaqi; Guo, Yi; Jiang, Zhongmin; Yang, Min; Li, Huaifang; Wang, Jianjun

    2015-04-01

    The aim of this study was to observe the effect of attenuated Salmonella typhi as a tumor-targeting delivery vector for multidrug-resistance gene (MDR1) small interfering RNA (siRNA). The cisplatin (DDP)-resistant ovarian cancer cell line SKOV-3/DDP was established by treatment with gradually increasing concentrations of cisplatin. MDR1 siRNA expression plasmid containing short hairpin RNA (shRNA) of MDR1 gene was constructed and transformed into attenuated Salmonella typhi strain  SL7207. SKOV-3/DDP cells were incubated with recombinant Salmonella and then subjected to analysis of MDR1 expression by real-time polymerase chain reaction and Western blot. SKOV-3/DDP tumor-bearing mice were established by subcutaneously injecting BALB/c nude mice with SKOV-3/DDP cells, and were orally inoculated with Salmonella carrying MDR1 siRNA plasmid and simultaneously injected intraperitoneally with cisplatin. Tumor growth and mouse survival were observed. Compared with parental cell line, the DDP-resistant SKOV-3/DDP cells expressed a much higher level of MDR1. The expression of MDR1 in SKOV-3/DDP cells infected with the Salmonella strain bearing MDR1 siRNA plasmid in vitro was detected to be downregulated and DDP tolerance of these cells was reversed. Tumor-bearing nude mice that were orally receiving recombinant Salmonella experienced a slow tumor growth and became more sensitive to DDP. Attenuated Salmonella typhi may represent a promising vector for in vivo administration of RNA interference therapy against malignant tumors. © 2014 The Authors. Journal of Obstetrics and Gynaecology Research © 2014 Japan Society of Obstetrics and Gynecology.

  4. Lose to win: marT pseudogenization in Salmonella enterica serovar Typhi contributed to the surV-dependent survival to H2O2, and inside human macrophage-like cells.

    Science.gov (United States)

    Ortega, A P; Villagra, N A; Urrutia, I M; Valenzuela, L M; Talamilla-Espinoza, A; Hidalgo, A A; Rodas, P I; Gil, F; Calderón, I L; Paredes-Sabja, D; Mora, G C; Fuentes, J A

    2016-11-01

    The difference in host range between Salmonella enterica serovar Typhimurium (S. Typhimurium) and Salmonella enterica serovar Typhi (S. Typhi) can be partially attributed to the gain of functions, to the loss of functions (i.e. pseudogenization), or to a combination of both processes. As previously reported, the loss of functions by pseudogenization may play a role in bacterial evolution, especially in host-restricted pathogens such as S. Typhi. The marT-fidL operon, located at the SPI-3, encodes the MarT transcriptional regulator and a hypothetical protein (i.e. FidL) with no significant similarities to known proteins, respectively. Even though predicted S. Typhimurium FidL exhibit 99.4% identity with S. Typhi FidL, marT has been annotated as a pseudogene in S. Typhi. In this work, we found that S. Typhi expressing S. Typhimurium marT-fidL exhibited an increased accumulation of reactive oxygen species (ROS), leading to a decreased survival in presence of H 2 O 2 . Moreover, we found that that the presence of a functional copy of S. Typhimurium marT-fidL in S. Typhi resulted in a repression of surV (STY4039), an ORF found in the S. Typhi SPI-3 but absent from S. Typhimurium SPI-3, that contribute to the resistance to H 2 O 2 by decreasing the accumulation of ROS. Finally, we observed that the presence of S. Typhimurium marT-fidL in S. Typhi negatively affected the survival inside macrophage-like cells, but not in epithelial cells, after 24h post infection. Therefore, this work provides evidence arguing that marT pseudogenization in Salmonella Typhi contributed to the surV-dependent survival against H 2 O 2 , and inside human macrophage-like cells. This is a good example of how the loss of functions (marT pseudogenization) and the gain of functions (presence of surV) might contribute to phenotypic changes improving virulence. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Polyamines are essential for virulence in Salmonella enterica serovar Gallinarum despite evolutionary decay of polyamine biosynthesis genes

    DEFF Research Database (Denmark)

    Schroll, Casper; Christensen, Jens P.; Christensen, Henrik

    2014-01-01

    -specificity coincides with accumulation of pseudogenes, indicating adaptation of host-restricted serovars to their narrow niches. Polyamines are small cationic amines and in Salmonella they can be synthesized through two alternative pathways directly from l-ornithine to putrescine and from l-arginine via agmatine...... to putrescine. The first pathway is not active in S. Gallinarum and S. Typhi, and this prompted us to investigate the importance of polyamines for virulence in S. Gallinarum. Bioinformatic analysis of all sequenced genomes of Salmonella revealed that pseudogene formation of the speC gene was exclusive for S....... Typhi and S. Gallinarum and happened through independent events. The remaining polyamine biosynthesis pathway was found to be essential for oral infection with S. Gallinarum since single and double mutants in speB and speE, encoding the pathways from agmatine to putrescine and from putrescine...

  6. Effect of reduced graphene oxide-hybridized ZnO thin films on the photoinactivation of Staphylococcus aureus and Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Teh, Swe Jyan; Yeoh, Soo Ling; Lee, Kian Mun; Lai, Chin Wei; Abdul Hamid, Sharifah Bee; Thong, Kwai Lin

    2016-08-01

    The immobilization of photocatalyst nanoparticles on a solid substrate is an important aspect for improved post-treatment separation and photocatalyst reactor design. In this study, we report the simple preparation of reduced graphene oxide (rGO)-hybridized zinc oxide (ZnO) thin films using a one-step electrochemical deposition, and investigated the effect of rGO-hybridization on the photoinactivation efficiency of ZnO thin films towards Staphylococcus aureus (S. aureus) and Salmonella enterica serovar Typhi (S. Typhi) as target bacterial pathogens. Field-emission scanning electron microscopy (FESEM) revealed the formation of geometric, hexagonal flakes of ZnO on the ITO glass substrate, as well as the incorporation of rGO with ZnO in the rGO/ZnO thin film. Raman spectroscopy indicated the successful incorporation of rGO with ZnO during the electrodeposition process. Photoluminescence (PL) spectroscopy indicates that rGO hybridization with ZnO increases the amount of oxygen vacancies, evidenced by the shift of visible PL peak at 650 to 500nm. The photoinactivation experiments showed that the thin films were able to reduce the bacterial cell density of Staph. aureus and S. Typhi from an initial concentration of approximately 10(8) to 10(3)CFU/mL within 15min. The rGO/ZnO thin film increased the photoinactivation rate for S. aureus (log[N/No]) from -5.1 (ZnO) to -5.9. In contrast, the application of rGO/ZnO thin film towards the photoinactivation of S. Typhi did not improve its photoinactivation rate, compared to the ZnO thin film. We may summarise that (1) rGO/ZnO was effective to accelerate the photoinactivation of S. aureus but showed no difference to improve the photoinactivation of S. Typhi, in comparison to the performance of ZnO thin films, and (2) the photoinactivation in the presence of ZnO and rGO/ZnO was by ROS damage to the extracellular wall. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Biosynthesis of silver nanoparticles using Myristica fragrans seed (nutmeg) extract and its antibacterial activity against multidrug-resistant (MDR) Salmonella enterica serovar Typhi isolates.

    Science.gov (United States)

    Balakrishnan, Senthilkumar; Sivaji, Ilakkia; Kandasamy, Selvam; Duraisamy, Senbagam; Kumar, Nachimuthu Senthil; Gurusubramanian, Guruswami

    2017-06-01

    Biosynthesis of nanoparticles has received increasing attention due its effective mode of action, eco-friendly preparation methodology, and less cytotoxicity. In the present study, silver nanoparticles (AgNPs) from aqueous seed extract of Myristica fragrans (nutmeg) were characterized. Gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of bioactive components acts as effective in reducing and capping agents for converting AgNO3 to AgNPs. The UV-Vis absorption spectrum of the biologically reduced reaction mixture showed the surface plasmon peak at 420 nm, which is the characteristic peak of AgNPs. The functional molecules present in the M. fragrans seed extract and their interaction with the AgNPs were identified by the Fourier transform infrared spectroscopy (FT-IR) analysis. X-ray diffraction (XRD) analysis confirmed the face-centered cubic crystalline structure of metallic silver nanoparticle and diameter was calculated using Scherrer's equation. Transmission electron microscope (TEM) image showed spherical shaped particles with an average size of 25 nm. The scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) confirmed the presence of elemental silver. The antibacterial activity of biosynthesized AgNPs was evaluated against multidrug-resistant (MDR) Salmonella enterica serovar Typhi (S. Typhi) according to agar well diffusion, MIC (minimum inhibitory concentration), and IC50 (inhibitory concentration 50%). The results confirm that bacterial growth was significantly reduced in a dose-dependent manner. Further, the cytotoxic effect of biosynthesized AgNPs on rat spleenocytes was analyzed. Thus, it is suggested that the nutmeg-biosynthesized AgNPs could be a lead drug and used effectively to control the MDR S. Typhi, thereby reducing public health issues and environmental pollution.

  8. Immunomodulatory and antimicrobial efficacy of Lactobacilli against enteropathogenic infection of Salmonella typhi: In-vitro and in-vivo study.

    Science.gov (United States)

    Mazaya, Basem; Hamzawy, Mohamed A; Khalil, Mahmoud A F; Tawkol, Wael M; Sabit, Hussein

    2015-12-01

    Salmonellosis-induced diarrhea, is one of the commonest cause of childhood mortality in developing countries. Using of probiotics is viewed as a promising means for reducing the pathogenic loads of bacterial infection. The current study aimed to evaluate the potential antimicrobial and immunomodulatory efficacy of isolated lactobacillus strains against the enteropathogenic effect of S. Typhi. Different Lactobacillus strains were isolated from 13 dairy products. Their antimicrobial activities were tested against different bacterial strains. Six groups of CD1 mice were treated for 8 days as follows: group (1) untreated control; group (2) was challenged with single inoculation S. typhi, and groups (3) and (4) were treated with Lactobacillus plantarum (LA5) or Lactobacillus paracsi (LA7) for 7 days, respectively. Groups (5) and (6) were challenged with S. typhi, and then treated with either LA5 or LA 7 for 7 days, respectively. Isolated Lactobacillus showed antimicrobial activity against wide range of bacterial strains. Salmonellosis showed high widal titer, induced significant disturbance of TNF and IL-1β, while sever changes of the histological patterns of the intestinal villi and hepatocytes have been illustrated. LA5 or LA7 succeeded to eradicate typhoid infection, restore the values of inflammatory cytokines to typical levels of control group, and improve histological pictures of intestinal and hepatic tissues. It can be concluded that lactobacilli are promising candidate in protection and eradication against bacterial infection induced by S. Typhi due to its antimicrobial, anti-inflammatory, and immunomodulatory activities. © The Author(s) 2015.

  9. An Evaluation of the Antimicrobial Potency of Lasianthera africana (BEAUV and Heinsia crinata (G. Taylor on Escherichia coli, Salmonella typhi, Staphylococcus aureus and Candida albicans

    Directory of Open Access Journals (Sweden)

    Andy, I. E.

    2008-01-01

    Full Text Available Information on the potency of many African medical plants against microorganisms is scanty, and in the current wave of antimicrobial resistance against chemotherapeutic drugs, there is need to search for plants that could be resistance-free and affordable. The objective of this study was to investigate the antimicrobial effects of the extracts of Lasianthera africana (E1 and Heinsia crinata (E2 in combination with chloramphenicol, on Escherichia coli, Salmonella typhi, Staphlococcus aureus and Candida albicans. For this purpose, the dilution sensitivity and disc diffusion techniques were respectively applied in determining minimum inhibitory concentration (MIC of the plant extracts, and the sensitivities of the organisms to the plant extracts and their combinations with chloramphenicol. L. africana and H. crinata showed very high antimicrobial activity against all the test organisms. In combination, the effect of E1 on E. coli and S. typhi was completely antagonized by that of E2, whereas additive effect on S. aureus and C. albicans was observed, indicating that the combination of E1 and E2 might be effective against gram positive pathogenic organisms. The combination of either plant extract with chloramphenicol produced synergistic effect on only C. albicans. The smaller MIC of E2 indicated greater effectivity than E1. It is concluded that the additive effect produced by the combination of the two plant extracts, and the synergic effect from the combination of any of the extracts with chloramphenicol, offer alternative therapy to gram positive bacterial infections and candidiasis respectively.

  10. Influence of subinhibitory-concentration (sub-MIC Cefetoxime on biofilm formation. SEM study of ESBL-producing Salmonella typhi

    Directory of Open Access Journals (Sweden)

    Rahul Narasanna, Manjunath Chavadi, Ajaykumar Oli

    2017-06-01

    Full Text Available Objectives: In the present study, we have analyzed ESBL-producing S. typhi’s capability in forming a significant amount of biofilm on plastic and glass surface, and the influence of cefetoxime on biofilm development at subinhibitory (Sub-MIC concentration. Methods: Nine strains of cefetoxime-mediated ESBL-producing S. typhi were used in the study. S. typhi formed biofilm on plastic and glass materials; it was demonstrated using micro titre plate (MTP and standard test tube methods. Comparative study of the influence of cefetoxime on biofilm formation in its MIC (128 µg/ml and at sub-MIC (64 µg/ml was demonstrated by microtitre plate method. The biofilm production was observed in SEM images, statistical analysis (ANOVA showed significant increase in cell surface and volume due to the influence of Cefetoxime. Results: Of the nine selected isolates, two S. typhi strains, namely BST 51 and BST 130, produced relatively strong biofilm in the presence of cefetoxime at sub-MIC level (64 µg/ml, comparatively weak biofilm formation at MIC level (128 µg/ml. Typical morphological changes were observed in cefetoxime-resistant strains, S. typhi BST 51 and BST 130, in comparison to cefetoxime-sensitive strain S. typhi BST 63 used as a control. We found an increase in surface and volume of a cell in response to cefetoxime and statistical data (ANOVA proved that resistant strains were significantly different from control strains. Conclusion: The above study clearly shows that cefetoxime at sub-MIC level efficiently induces biofilm formation and promotes changes in morphology of the cell. J Microbiol Infect Dis 2017; 7(2: 67-75

  11. Lisotipia Vi e classificação bioquímica de amostras de Salmonella typhi isoladas no Estado da Guanabara

    Directory of Open Access Journals (Sweden)

    Ernesto Hofer

    1972-01-01

    Full Text Available Na presente investigação, os autores estudaram a distribuição dos lisótipos Ví, tipos fermentativos e tetrationato-redutase, em 110 amostras de Salmonella typhi, isoladas no período de 1966-1970, no Estado da Guanabara. Para estabelecer uma melhor configuração epidemiológica dos resultados, fez-se a divisão do Estado em seis regiões, respectivamente as zonas Sul, Centro, Norte, da E. F. Central, da E. F. Leopoldina e rural. Obteve-se no cômputo geral, a prevalência dos lisótipos A (59,09%, E 1a (19,09%, T (7,27% e amostras Vi negativas (4,55%. Verificou-se ainda, que cerca de 70,90% das culturas foram classificadas no tipo fermentativo II, segundo esquema de Kristensen e 95,42% das amostras revelaram-se positivas na prova de detecção da tetrationato-redutase.In the present investigation, the authors have studied the distribution of lysotypes, fermentative types and tetrathionate- reductase in 110 strains of Salmonella typhi, isolated from hemocultures during the period 1966-1970, in the State of Guanabara, Brazil. In order to establish a better epidemiological configuration of the results, this study has been divided and effected in the six following regions: south, middle, north at the Central Railway, the Leopoldina Railway and the rural district. As a general outcome a predomination of the lysotypes A, E 1a, T and negative Vi strains has been obtained. It has stil been verified that 70,90% of the cultures were classified fermentative type Ii and that 95,42% of the strains were tetrathionate-reductase positives.

  12. Screening of some medicinal plants used in south-west Nigerian traditional medicine for anti-Salmonella typhi activity.

    Science.gov (United States)

    Akinyemi, K O; Mendie, U E; Smith, S T; Oyefolu, A O; Coker, A O

    2005-01-01

    Ten Nigerian medicinal plants used traditionally for the treatment of several ailments of both microbial and non-microbial origins were tested on multi-drug resistant S. typhi (MDR) strains of which six of them were active. The results revealed that both the aqueous and ethanol extracts of Terminalia avicennioides, Momordica balsamina, Combretum paniculatum and Trema guineensis were effective on the MDR-S. typhi strains with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values ranging from 9.60 to 14 mcg/ml and 24 to 33 mcg/ml, respectively. Whereas, only the aqueous extracts of Morinda lucida and Ocimum gratissimum were found to be active against this pathogen with MIC and MBC values of 9.60 and 24 mcg/ml for M. lucida, 40 and 55 mcg/ml for O. gratissimum, respectively. There was no statistical significant difference (P > 0.05) between the activity of each plant extract and the decoctions prepared from them. All the six active plants showed positive reactions to alkaloids, tannins, flavonoids and anthraquinones but in variable degrees. All but M. balsamina, indicated the presence of saponin.

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

    Directory of Open Access Journals (Sweden)

    Rui Figueiredo

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

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

    Science.gov (United States)

    Figueiredo, Rui; Card, Roderick; Nunes, Carla; AbuOun, Manal; Bagnall, Mary C; Nunez, Javier; Mendonça, Nuno; Anjum, Muna F; da Silva, Gabriela Jorge

    2015-01-01

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

  15. Knowledge, attitude and practice of hygiene and sanitation in a Burundian refugee camp: implications for control of a Salmonella typhi outbreak.

    Science.gov (United States)

    Nahimana, Marie-Rosette; Ngoc, Candide Tran; Olu, Olushayo; Nyamusore, Jose; Isiaka, Ayodeji; Ndahindwa, Vedaste; Dassanayake, Lakruwan; Rusanganwa, André

    2017-01-01

    A Salmonella typhi outbreak was reported in a Burundian refugee camp in Rwanda in October 2015. Transmission persisted despite increased hygiene promotion activities and hand-washing facilities instituted to prevent and control the outbreak. A knowledge, attitude and practice (KAP) study was carried out to assess the effectiveness of ongoing typhoid fever preventive interventions. A cross-sectional survey was conducted in Mahama Refugee Camp of Kirehe District, Rwanda from January to February 2016. Data were obtained through administration of a structured KAP questionnaire. Descriptive, bivariate and multivariate analysis was performed using STATA software. A total of 671 respondents comprising 264 (39.3%) males and 407 (60.7%) females were enrolled in the study. A comparison of hand washing practices before and after institution of prevention and control measures showed a 37% increase in the proportion of respondents who washed their hands before eating and after using the toilet (p practices. The findings of this study underline the need for bolstering up health education and hygiene promotion activities in Mahama and other refugee camp settings.

  16. Immunization with the conjugate vaccine Vi-CRM₁₉₇ against Salmonella typhi induces Vi-specific mucosal and systemic immune responses in mice.

    Science.gov (United States)

    Fiorino, Fabio; Ciabattini, Annalisa; Rondini, Simona; Pozzi, Gianni; Martin, Laura B; Medaglini, Donata

    2012-09-21

    Typhoid fever is a public health problem, especially among young children in developing countries. To address this need, a glycoconjugate vaccine Vi-CRM₁₉₇, composed of the polysaccharide antigen Vi covalently conjugated to the non-toxic mutant of diphtheria toxin CRM₁₉₇, is under development. Here, we assessed the antibody and cellular responses, both local and systemic, following subcutaneous injection of Vi-CRM₁₉₇. The glycoconjugate elicited Vi-specific serum IgG titers significantly higher than unconjugated Vi, with prevalence of IgG1 that persisted for at least 60 days after immunization. Vi-specific IgG, but not IgA, were present in intestinal washes. Lymphocytes proliferation after restimulation with Vi-CRM₁₉₇ was observed in spleen and mesenteric lymph nodes. These data confirm the immunogenicity of Vi-CRM₁₉₇ and demonstrate that the vaccine-specific antibody and cellular immune responses are present also in the intestinal tract, thus strengthening the suitability of Vi-CRM₁₉₇ as a promising candidate vaccine against Salmonella Typhi. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. Salmonella Sepsis in African Children

    African Journals Online (AJOL)

    Infection with both Salmonella typhiand non-typhi salmonella. (NTS) is common among children in many African countries. Salmonella typhi predominates among older children and adults with the typical localising features of enteric fever. Nontyphoid salmonellae species are more often reported among children under 5 ...

  18. Inhibitory Activity of Lactid Acid Bacteria Isolated from Tape Waterlily Seed to Enteric Pathogenic Bacteria (Vibrio cholera, Salmonella typhi, Shigella disentri, and E.coli and Its’ Susceptibility to Antibiotic, Bile Salt and Acidic Condition

    Directory of Open Access Journals (Sweden)

    Iin Khusnul Khotimah

    2012-03-01

    Full Text Available The aim of this research was to observe inhibitory activity of LAB isolated from tape waterlily seed to enteric pathogenic bacteria (Vibrio cholera, Salmonella typhi, Shigella disentri, E.coli ATCC 25922 and it’s susceptibility to antibiotic, in bile salt and under acidic condition. Microbia in the tape ( a fermented product of waterlily seed to showed were Streptococcus thermophilus (IKH-1, Pediococcus pentosaceus (IKH-2 and Leuconostoc mesentroides (IKH-8. Streptococcus thermophillus showed inhibition against the growth of Shigella disentri with inhibition zones 16,28 mm, but did not against the growth of V. Cholera, S. typhi, E.coli. Pediococcus pentosaceus inhibit Vibrio cholera, dan Salmonella thypi with inhibition zones 18,59 mm dan 7,91 mm. So that, Leuconostoc mesenteroides inhibit Salmonella thypi with zones inhibits average 8,25 mm. Chloramfenicol at 0.05 mg concentrations did not show inhibition against the growth of isolated Streptococcus thermophillus, Pediococcus pentosaceus and Leuconostoc mesentroides. These isolates could survive too in bile salt (2% and acidified media (pH 3.   Keyword : The tape of  waterlily seed, LAB, probiotic and enteric pathogenic   KEMAMPUAN PENGHAMBATAN BAKTERI ASAM LAKTAT DARI TAPE BIJI TERATAI TERHADAP PATOGENIK ENTERIK (VIBRIO CHOLERA, SALMONELLA THYPI, SHIGELLA DISENTRI, E. COLI, ANTIBIOTIK, KETAHANANNYA TERHADAP BILE SALT DAN ASAM   ABSTRAK   Penelitian ini bertujuan untuk menguji kemampuan penghambatan bakteri asam laktat yang diisolasi dari tape biji teratai terhadap patogenik enterik (Vibrio cholera, Salmonella thypi, Shigella disentri, E. Coli ATCC 25922, antibiotik, bile salt dan asam. Jenis bakteri yang diketahui tumbuh selama fermentasi tape biji teratai adalah Streptococcus thermopilus (IKH-1, Pediococcus pentosaceus(IKH-2, dan Leuconostoc mesentroides (IKH-8. Pengamatan terhadap uji penghambatan patogenik enterik (Vibrio cholera, Salmonella thypi, Shigella disentri, dan E. Coli ATCC

  19. Clinicopathological profile of salmonella typhi and paratyphi infections presenting as Fever of unknown origin in a tropical country.

    Science.gov (United States)

    Iqbal, Nayyar; Basheer, Aneesh; Mookkappan, Sudhagar; Ramdas, Anita; Varghese, Renu G'Boy; Padhi, Somanath; Shrimanth, Bhairappa; Chidambaram, Saranya; Anandhalakshmi, S; Kanungo, Reba

    2015-01-01

    Enteric fever, a common infection in the tropics and endemic to India, often manifests as an acute febrile illness. However, presentation as fever of unknown origin (FUO) is not uncommon in tropical countries. We aim to describe the clinical, laboratory and pathological features of patients hospitalized with fever of unknown origin and diagnosed as enteric fever. All culture proven cases of enteric fever were analyzed retrospectively over a period of three years from January 2011 to December 2013. Seven of 88 (8%) cases with enteric fever presented as FUO. Abdominal pain was the most common symptom besides fever. Relative bradycardia and splenomegaly were uncommon. Thrombocytopenia was the most common haematological abnormality while leucopenia was rare. Transaminase elevation was almost universal. S. Typhi and S. Paratyphi A were isolated from six cases and one case respectively. Yield of organisms from blood culture was superior to that of bone marrow aspirate. Multiple granulomas were identified in 4 out of 6 (67%) of the bone marrows studied, including that due to S. Paratyphi A and histiocytic hemophagocytosis was noted in two cases. FUO is a relatively common manifestation of enteric fever in the tropics. Clinical and laboratory features may be atypical in such cases, including absence of relative bradycardia, leucopenia, and presence of thrombocytopenia, bicytopenia or pancytopenia. In addition, in endemic countries, enteric fever should be considered as a differential diagnosis, next to tuberculosis, in the evaluation of bone marrow granulomas in cases with FUO and culture correlation should be mandatory.

  20. Crl binds to domain 2 of σ(S) and confers a competitive advantage on a natural rpoS mutant of Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Monteil, Véronique; Kolb, Annie; Mayer, Claudine; Hoos, Sylviane; England, Patrick; Norel, Françoise

    2010-12-01

    The RpoS sigma factor (σ(S)) is the master regulator of the bacterial response to a variety of stresses. Mutants in rpoS arise in bacterial populations in the absence of stress, probably as a consequence of a subtle balance between self-preservation and nutritional competence. We characterized here one natural rpoS mutant of Salmonella enterica serovar Typhi (Ty19). We show that the rpoS allele of Ty19 (rpoS(Ty19)) led to the synthesis of a σ(S)(Ty19) protein carrying a single glycine-to-valine substitution at position 282 in σ(S) domain 4, which was much more dependent than the wild-type σ(S) protein on activation by Crl, a chaperone-like protein that increases the affinity of σ(S) for the RNA polymerase core enzyme (E). We used the bacterial adenylate cyclase two-hybrid system to demonstrate that Crl bound to residues 72 to 167 of σ(S) domain 2 and that G282V substitution did not directly affect Crl binding. However, this substitution drastically reduced the ability of σ(S)(Ty19) to bind E in a surface plasmon resonance assay, a defect partially rescued by Crl. The modeled structure of the Eσ(S) holoenzyme suggested that substitution G282V could directly disrupt a favorable interaction between σ(S) and E. The rpoS(Ty19) allele conferred a competitive fitness when the bacterial population was wild type for crl but was outcompeted in Δcrl populations. Thus, these results indicate that the competitive advantage of the rpoS(Ty19) mutant is dependent on Crl and suggest that crl plays a role in the appearance of rpoS mutants in bacterial populations.

  1. RpoE promotes invasion and intracellular survival by regulating SPI-1 and SPI-2 in Salmonella enterica serovar Typhi.

    Science.gov (United States)

    Zhang, Haifang; Jia, Yanwei; Xie, Xiaofang; Wang, Min; Zheng, Yi; Xu, Shungao; Zhang, Wei; Wang, Qiang; Huang, Xinxiang; Du, Hong

    2016-08-01

    To demonstrate the role of RpoE during the later stage of hyperosmotic stress in Salmonella. Expressions of SPI-1 and SPI-2 under hyperosmotic stress for 120 min were investigated by a microarray, and the invasion and intracellular survival of wild-type and ΔrpoE strains were compared. The global differential expression of bacterial proteins between the wild-type and ΔrpoE strains was examined after 120 min of hyperosmotic stress. SPI-1 and SPI-2 were repressed, and the invasion and intracellular survival were defected in the ΔrpoE strain. Thirteen bacterial-associated proteins and 11 secreted proteins differed significantly between the wild-type and ΔrpoE strains. RpoE may promote invasion and intracellular survival by regulating the expression of SPI-1 and SPI-2.

  2. Epitope tagging of chromosomal genes in Salmonella.

    Science.gov (United States)

    Uzzau, S; Figueroa-Bossi, N; Rubino, S; Bossi, L

    2001-12-18

    We have developed a simple and efficient procedure for adding an epitope-encoding tail to one or more genes of interest in the bacterial chromosome. The procedure is a modification of the gene replacement method of Datsenko and Wanner [Datsenko, K. A. & Wanner, B. L. (2000) Proc. Natl. Acad. Sci. USA 97, 6640-6645]. A DNA module that begins with the epitope-encoding sequence and includes a selectable marker is amplified by PCR with primers that carry extensions (as short as 36 nt) homologous to the last portion of the targeted gene and to a region downstream from it. Transformation of a strain expressing bacteriophage lambda red functions yields recombinants carrying the targeted gene fused to the epitope-encoding sequence. The resulting C-terminal-tagged protein can be identified by standard immuno-detection techniques. In an initial application of the method, we have added the sequences encoding the FLAG and 3xFLAG and influenza virus hemagglutinin epitopes to various genes of Salmonella enterica serovar Typhimurium, including putative and established pathogenic determinants present in prophage genomes. Epitope fusion proteins were detected in bacteria growing in vitro, tissue culture cells, and infected mouse tissues. This work identified a prophage locus specifically expressed in bacteria growing intracellularly. The procedure described here should be applicable to a wide variety of Gram-negative bacteria and is particularly suited for the study of intracellular pathogens.

  3. SALMONELLA TYPHI SPONDYUTIS: AN UNUSUAL ...

    African Journals Online (AJOL)

    Her low back pain had become disabling to the extent that she could not walk. On admission she was frail, emaciated and malnourished but apyrexial. ... specific treatment but the low back pain became incapacitating. Anterior spinal fusion resulted in relief from pain. Since the patient suffered from low back pain for 15.

  4. Treatment of enteric fever in children on the basis of current trends of antimicrobial susceptibility of Salmonella enterica serovar typhi and paratyphi A

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    Manchanda V

    2006-01-01

    Full Text Available Purpose: Recent reports indicate decreased susceptibility of S. typhi to fluoroquinolones, especially ciprofloxacin. Chloramphenicol has been suggested as first line therapy of enteric fever in many studies. This is a prospective study that describes the trends of antimicrobial susceptibility of S. typhi and S. paratyphi A causing bacteraemia in children and reports therapeutic failure to ciprofloxacin and evaluates the possible use of chloramphenicol, ampicillin, ciprofloxacin and third generation cephalosporins as first line therapy in the treatment of enteric fever in children. Methods: The present study was conducted from April 2004 to March 2005 in a superspeciality children hospital at New Delhi. A total of 56 S. typhi and five S. paratyphi A isolates were obtained among the 673 blood cultures performed. Antimicrobial testing was done using disk diffusion technique (NCCLS method for 13 antimicrobials and MICs were calculated for ampicillin, ciprofloxacin, chloramphenicol and cefotaxime. Analysis of data was done using WHONET software. Results: All 56 isolates of S. typhi were sensitive to amoxycillin+clavulanate, gentamicin, cefixime, cefotaxime and ceftazidime. Multidrug resistance (MDR, resistance to three drugs was seen in 22 cases (39% and resistance to five drugs was seen in 12 cases (21%. Only two isolates were resistant to chloramphenicol (3%. MIC 90 for ampicillin, chloramphenicol, ciprofloxacin and cefotaxime were 1.0 mg/ml, 4.0 mg/ml, 64 mg/ml and 0.125 mg/ml respectively. All S. paratyphi A isolates were sensitive to ampicillin and chloramphenicol and resistant to nalidixic acid.MIC distribution data for chloramphenicol revealed elevated MIC but still in susceptible range. Conclusions: There is an urgent need for further clinical studies to evaluate response to chloramphenicol in such cases. Antimicrobial susceptibility data and MIC distribution favour use of ampicillin as a drug of choice for the treatment of enteric fever

  5. Gene co-expression network analysis identifies porcine genes associated with variation in Salmonella shedding.

    Science.gov (United States)

    Kommadath, Arun; Bao, Hua; Arantes, Adriano S; Plastow, Graham S; Tuggle, Christopher K; Bearson, Shawn M D; Guan, Le Luo; Stothard, Paul

    2014-06-09

    Salmonella enterica serovar Typhimurium is a gram-negative bacterium that can colonise the gut of humans and several species of food producing farm animals to cause enteric or septicaemic salmonellosis. While many studies have looked into the host genetic response to Salmonella infection, relatively few have used correlation of shedding traits with gene expression patterns to identify genes whose variable expression among different individuals may be associated with differences in Salmonella clearance and resistance. Here, we aimed to identify porcine genes and gene co-expression networks that differentiate distinct responses to Salmonella challenge with respect to faecal Salmonella shedding. Peripheral blood transcriptome profiles from 16 pigs belonging to extremes of the trait of faecal Salmonella shedding counts recorded up to 20 days post-inoculation (low shedders (LS), n = 8; persistent shedders (PS), n = 8) were generated using RNA-sequencing from samples collected just before (day 0) and two days after (day 2) Salmonella inoculation. Weighted gene co-expression network analysis (WGCNA) of day 0 samples identified four modules of co-expressed genes significantly correlated with Salmonella shedding counts upon future challenge. Two of those modules consisted largely of innate immunity related genes, many of which were significantly up-regulated at day 2 post-inoculation. The connectivity at both days and the mean gene-wise expression levels at day 0 of the genes within these modules were higher in networks constructed using LS samples alone than those using PS alone. Genes within these modules include those previously reported to be involved in Salmonella resistance such as SLC11A1 (formerly NRAMP1), TLR4, CD14 and CCR1 and those for which an association with Salmonella is novel, for example, SIGLEC5, IGSF6 and TNFSF13B. Our analysis integrates gene co-expression network analysis, gene-trait correlations and differential expression to provide new

  6. Efficacy and immunogenicity of a Vi-tetanus toxoid conjugate vaccine in the prevention of typhoid fever using a controlled human infection model of Salmonella Typhi: a randomised controlled, phase 2b trial.

    Science.gov (United States)

    Jin, Celina; Gibani, Malick M; Moore, Maria; Juel, Helene B; Jones, Elizabeth; Meiring, James; Harris, Victoria; Gardner, Jonathan; Nebykova, Anna; Kerridge, Simon A; Hill, Jennifer; Thomaides-Brears, Helena; Blohmke, Christoph J; Yu, Ly-Mee; Angus, Brian; Pollard, Andrew J

    2017-12-02

    Salmonella enterica serovar Typhi (S Typhi) is responsible for an estimated 20 million infections and 200 000 deaths each year in resource poor regions of the world. Capsular Vi-polysaccharide-protein conjugate vaccines (Vi-conjugate vaccines) are immunogenic and can be used from infancy but there are no efficacy data for the leading candidate vaccine being considered for widespread use. To address this knowledge gap, we assessed the efficacy of a Vi-tetanus toxoid conjugate vaccine using an established human infection model of S Typhi. In this single-centre, randomised controlled, phase 2b study, using an established outpatient-based human typhoid infection model, we recruited healthy adult volunteers aged between 18 and 60 years, with no previous history of typhoid vaccination, infection, or prolonged residency in a typhoid-endemic region. Participants were randomly assigned (1:1:1) to receive a single dose of Vi-conjugate (Vi-TT), Vi-polysaccharide (Vi-PS), or control meningococcal vaccine with a computer-generated randomisation schedule (block size 6). Investigators and participants were masked to treatment allocation, and an unmasked team of nurses administered the vaccines. Following oral ingestion of S Typhi, participants were assessed with daily blood culture over a 2-week period and diagnosed with typhoid infection when meeting pre-defined criteria. The primary endpoint was the proportion of participants diagnosed with typhoid infection (ie, attack rate), defined as persistent fever of 38°C or higher for 12 h or longer or S Typhi bacteraemia, following oral challenge administered 1 month after Vi-vaccination (Vi-TT or Vi-PS) compared with control vaccination. Analysis was per protocol. This trial is registered with ClinicalTrials.gov, number NCT02324751, and is ongoing. Between Aug 18, 2015, and Nov 4, 2016, 112 participants were enrolled and randomly assigned; 34 to the control group, 37 to the Vi-PS group, and 41 to the Vi-TT group. 103 participants

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

    DEFF Research Database (Denmark)

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

    2012-01-01

    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...... 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...

  8. PCR-RFLP Analysis of a fliC Gene Fragment in Avian Salmonella Isolates

    Directory of Open Access Journals (Sweden)

    Zohreh Ebrahimvandi

    2014-07-01

    Full Text Available Background: Salmonella are a genus of zoonotic bacteria of worldwide economic and health importance. Members of Salmonella enterica subspecies enterica are mainly associated with warm-blooded vertebrates and are usually transmitted by ingestion of food or watercontaminated by infected feces. Objectives: The aim of this study was to apply a PCR-RFLP method based on the fliC gene to identify the serotypes of Salmonella isolates from Karaj, Iran. Materials and Methods: A total of 30 Salmonella isolates were serotyped by specific antisera. For the PCR-RFLP method based on the fliC gene, extracted DNA was used as the template for amplifying the fliC gene (1500 bp using specific primers. PCR products were subjected to digestion using HhaI restriction endonuclease. Results: This study determined 30 serotypes as Salmonella durban (56.6%, Salmonella uno (23.3%, Salmonella enteritidis (3.3%, Salmonella tinda (3.3%, Salmonella mjimweme (3.3%, Salmonella Thompson (3.3%, Salmonella sIIO8 (3.3 % and Salmonella sIIO7 (3.3%. Observations indicated that HhaI is able to discriminate Salmonella tinda and Salmonella thompson, yet Salmonella enteritidis, Salmonella durban and Salmonella mjimweme had the same pattern with this enzyme. Also Salmonella sIIO8, Salmonella sIIO7 and Salmonella uno showed the same pattern. Thus, regarding the size and the number of resulting fragments from this enzyme, four patterns were obtained for HhaI. Conclusion: A large number of Salmonella serotypes need to be analyzed by the PCR-RFLP method and different enzymes must be used to give reliable results.

  9. Identification of Specific Gene Sequences Conserved in Contemporary Epidemic Strains of Salmonella enterica▿ †

    OpenAIRE

    Kang, Min-Su; Besser, Thomas E.; Hancock, Dale D.; Porwollik, Steffen; McClelland, Michael; Call, Douglas R.

    2006-01-01

    Genetic elements specific to recent and contemporary epidemic strains of Salmonella enterica were identified using comparative genomic analysis. Two epidemic multidrug-resistant (MDR) strains, MDR Salmonella enterica serovar Typhimurium definitive phage type 104 (DT104) and cephalosporin-resistant MDR Salmonella enterica serovar Newport, and an epidemic pansusceptible strain, Salmonella serovar Typhimurium DT160, were subjected to Salmonella gene microarray and suppression subtractive hybridi...

  10. Hydrogen-Stimulated Gene Expression by Salmonella enterica serovar Typhimurium in a Carbon Limited Environment

    Science.gov (United States)

    Background: Salmonella enterica serovar Typhimurium can utilize molecular hydrogen for growth and amino acid transport during anaerobic growth in a carbon limited environment. In this study we identified hydrogen-stimulated gene expression changes contributing to Salmonella survival. Methods: Micr...

  11. Gene co-expression network analysis identifies porcine genes associated with variation in Salmonella shedding

    OpenAIRE

    Kommadath, Arun; Bao, Hua; Arantes, Adriano S.; Plastow, Graham S; Tuggle, Christopher K.; Bearson, Shawn MD; Luo Guan, Le; Stothard, Paul

    2014-01-01

    Background Salmonella enterica serovar Typhimurium is a gram-negative bacterium that can colonise the gut of humans and several species of food producing farm animals to cause enteric or septicaemic salmonellosis. While many studies have looked into the host genetic response to Salmonella infection, relatively few have used correlation of shedding traits with gene expression patterns to identify genes whose variable expression among different individuals may be associated with differences in ...

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

    Directory of Open Access Journals (Sweden)

    Muhammed Shafeekh Muyyarikkandy

    2017-11-01

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

  13. Determinación de la relación clonal de los aislamientos de Salmonella typhi recuperados en el programa de vigilancia por el laboratorio de EDA en Colombia durante el período 1997-2003 mediante la electroforesis en campo pulsado

    Directory of Open Access Journals (Sweden)

    Hidalgo Marylin

    2005-07-01

    Full Text Available La tipificación molecular de Salmonella typhi y paratyphi es necesaria para complementar los análisis epidemiológicos de las fiebres entéricas en Colombia. El objetivo del trabajo fue establecer la relación genética de los aislamientos de Salmonella typhi y paratyphi recuperados durante el período 1997-2003, en varias regiones del país. Se utilizaron 58 aislamientos de S. typhi, tres de S. paratyphi A, dos de S. paratyphi B y 1 de S. paratyphi C, recibidos por el programa de vigilancia de EDA del Instituto Nacional de Salud (INS. Para determinar la relación genética se empleó la electroforesis en gel de campo pulsado y se calculó un dendrograma; para evaluar la asociación entre los patrones electroforéticos obtenidos y las características de los aislamientos, como datos
    demográficos de los pacientes, tipo de muestra, año y procedencia, se utilizó un análisis de correspondencias. Los datos de S. typhi se analizaron en conjunto con 15  islamientos tipificados anteriormente en el INS. Se obtuvieron 36 patrones electroforéticos, 31/36 (86,1%, representados en 68/73 (93,1% aislamientos, estuvieron relacionados genéticamente. Los patrones S.Ty0001 y S.Ty0012 predominaron en el país. En el análisis de correspondencias, no se encontró asociación entre los patrones electroforéticos y las características de los aislamientos. Los aislamientos de S. paratyphi A estuvieron relacionados genéticamente mientras que los de S. paratyphi B no guardaron relación. A pesar de la considerable heterogeneidad de los aislamientos colombianos de Salmonella typhi, definida por el número de patrones electroforéticos, la mayoría de ellos guardan una relación genética, sugiriendo un origen común de los mismos.

  14. Salmonella typhimurium activates virulence gene transcription within acidified macrophage phagosomes.

    Science.gov (United States)

    Alpuche Aranda, C M; Swanson, J A; Loomis, W P; Miller, S I

    1992-01-01

    Survival of Salmonella typhimurium within macrophage phagosomes requires the coordinate expression of bacterial gene products. This report examines the contribution of phagosomal pH as a signal for expression of genes positively regulated by the S. typhimurium virulence regulators PhoP and PhoQ. Several hours after bacterial phagocytosis by murine bone marrow-derived macrophages, PhoP-activated gene transcription increased 50- to 77-fold. In contrast, no difference in PhoP-activated gene expression was observed after infection of cultured epithelial cells, suggesting that the membrane sensor PhoQ recognized signals unique to macrophage phagosomes. The increase in PhoP-regulated gene expression was abolished when macrophage culture medium contained NH4Cl or chloroquine, weak bases that raise the pH of acidic compartments. Measurements of pH documented that S. typhimurium delayed and attenuated acidification of its intracellular compartment. Phagosomes containing S. typhimurium required 4-5 hr to reach pH < 5.0. In contrast, within 1 hr vacuoles containing heat-killed bacteria were measured at pH < 4.5. The eventual acidification of phagosomes to pH < 5.0 correlated with the period of maximal PhoP-dependent gene expression. These observations implicate phagosome acidification as an intracellular inducer of PhoP-regulated gene expression and suggest that Salmonella survival is dependent on its ability to attenuate phagosome acidification. Images PMID:1438196

  15. Association of twelve candidate gene polymorphisms and response to challenge with Salmonella enteritidis in poultry

    NARCIS (Netherlands)

    Kramer, J.; Malek, M.; Lamont, S.J.

    2003-01-01

    Breeding for disease resistance to Salmonella enteritidis (SE) could be an effective approach to control Salmonella in poultry. The candidate gene approach is a useful method to investigate genes that are involved in genetic resistance. In this study, 12 candidate genes that are involved in the

  16. Epidemic increase in Salmonella bloodstream infection in children, Bwamanda, the Democratic Republic of Congo.

    Science.gov (United States)

    Phoba, M-F; De Boeck, H; Ifeka, B B; Dawili, J; Lunguya, O; Vanhoof, R; Muyembe, J-J; Van Geet, C; Bertrand, S; Jacobs, J

    2014-01-01

    Salmonella enterica is the leading cause of bloodstream infection in children in sub-Saharan Africa, but few data are available from Central-Africa. We documented during the period November 2011 to May 2012 an epidemic increase in invasive Salmonella bloodstream infections in HGR Bwamanda, a referral hospital in Equateur Province, DR Congo. Salmonella spp. represented 90.4 % (103 out of 114) of clinically significant blood culture isolates and comprised Salmonella Typhimurium (54.4 %, 56 out of 103), Salmonella Enteritidis (28.2 %, 29 out of 103) and Salmonella Typhi (17.5 %, 18 out of 103), with Salmonella Enteritidis accounting for most of the increase. Most (82 out of 103, 79.6 %) isolates were obtained from children Salmonella Typhimurium and Salmonella Enteritidis were 14 months (14 days to 64 years) and 19 months (3 months to 8 years) respectively. Clinical presentation was non-specific; the in-hospital case fatality rate was 11.1 %. More than two thirds (69.7 %, 53 out of 76) of children Salmonella isolates as well as 6/18 (33.3 %) Salmonella Typhi isolates were multidrug resistant (i.e. resistant to the first-line oral antibiotics amoxicillin, trimethoprim-sulfamethoxazole and chloramphenicol), one (1.0 %) Salmonella Typhimurium had decreased ciprofloxacin susceptibility owing to a point mutation in the gyrA gene (Gly81Cys). Multilocus variable-number tandem-repeat (MLVA) analysis of the Salmonella Enteritidis isolates revealed closely related patterns comprising three major and four minor profiles, with differences limited to one out of five loci. These data show an epidemic increase in clonally related multidrug-resistant Salmonella bloodstream infection in children in DR Congo.

  17. The Role of the spv Genes in Salmonella Pathogenesis

    Directory of Open Access Journals (Sweden)

    Donald G. Guiney

    2011-06-01

    Full Text Available Salmonella strains cause three main types of diseases in people: gastroenteritis, enteric (typhoid fever, and non-typhoid extra-intestinal disease with bacteremia. Genetic analysis indicates that each clinical syndrome requires distinct sets of virulence genes, and Salmonella isolates differ in their constellation of virulence traits. The spv locus is strongly associated with strains that cause non-typhoid bacteremia, but are not present in typhoid strains. The spv region contains three genes required for the virulence phenotype in mice: the positive transcriptional regulator spvR and two structural genes spvB and spvC. SpvB and SpvC are translocated into the host cell by the SPI-2 type-three secretion system. SpvB prevents actin polymerization by ADP-ribosylation of actin monomers, while SpvC has phosphothreonine lyase activity and has been shown to inhibit MAP kinase signaling. The exact mechanisms by which SpvB and SpvC act in concert to enhance virulence are still unclear. SpvB exhibits a cytotoxic effect on host cells and is required for delayed cell death by apoptosis following intracellular infection. Strains isolated from systemic infections of immune compromised patients, particularly HIV patients, usually carry the spv locus, strongly suggesting that CD4 T cells are required to control disease due to Salmonella that are spv positive. This association is not seen with typhoid fever, indicating that the pathogenesis and immunology of typhoid have fundamental differences from the syndrome of non-typhoid bacteremia.

  18. Study on the promotion of bacterial biofilm formation by a Salmonella conjugative plasmid and the underlying mechanism.

    Directory of Open Access Journals (Sweden)

    Zhen Liu

    Full Text Available To investigate the effect of the pRST98 plasmid, originally isolated from Salmonella enterica serovar Typhi (S. Typhi, on biofilm (BF formation, we carried out in vitro experiments using S. Typhi, Salmonella enterica serovar Typhimurium (S. Typhimurium and Escherichia coli (E. coli. We further explored the effects of pRST98 in vivo by establishing two animal models, a tumor-bearing mouse model and a mouse urethral catheter model. Moreover, we examined the relationship between the quorum-sensing (QS system and pRST98-mediated BF formation. These studies showed that pRST98 enhanced BF formation in different bacteria in vitro. In both animal models, pRST98 promoted BF formation and caused more severe pathological changes. It was previously reported that Salmonella senses exogenous N-acylhomoserine lactones (AHLs through the regulatory protein SdiA and regulates the expression of genes including the virulence gene rck, which is located on the virulence plasmid of some serotypes of Salmonella. In this study, we confirmed the locus of the rck gene on pRST98 and found that AHLs increased rck expression in pRST98-carrying strains, thereby enhancing bacterial adherence, serum resistance and bacterial BF formation. In conclusion, the Salmonella conjugative plasmid pRST98 promotes bacterial BF formation both in vitro and in vivo, and the mechanism may relate to the AHL-SdiA-Rck signaling pathway.

  19. PhoP-Induced Genes within Salmonella Pathogenicity Island 1

    Science.gov (United States)

    Aguirre, Andrés; Cabeza, María Laura; Spinelli, Silvana V.; McClelland, Michael; García Véscovi, Eleonora; Soncini, Fernando C.

    2006-01-01

    The invasive pathogen Salmonella enterica has evolved a sophisticated device that allows it to enter nonphagocytic host cells. This process requires the expression of Salmonella pathogenicity island 1 (SPI-1), which encodes a specialized type III protein secretion system (TTSS). This TTSS delivers a set of effectors that produce a marked rearrangement of the host cytoskeleton, generating a profuse membrane ruffling at the site of interaction, driving bacterial entry. It has been shown that the PhoP/PhoQ two-component system represses the expression of the SPI-1 machinery by down-regulating the transcription of its master regulator, HilA. In this work, we reveal the presence of a PhoP-activated operon within SPI-1. This operon is composed of the orgB and orgC genes, which encode a protein that interacts with the InvC ATPase and a putative effector protein of the TTSS, respectively. Under PhoP-inducing conditions, expression of this operon is directly activated by the phosphorylated form of the response regulator, which recognizes a PhoP box located at the −35 region relative to the transcription start site. Additionally, under invasion-inducing conditions, orgBC expression is driven both by the prgH promoter, induced by the SPI-1 master regulator HilA, and by the directly controlled PhoP/PhoQ promoter. Together, these results indicate that in contrast to the rest of the genes encompassed in the SPI-1 locus, orgBC is expressed during and after Salmonella entry into its host cell, and they suggest a role for the products of this operon after host cell internalization. PMID:16980492

  20. Global Screening ofSalmonella entericaSerovar Typhimurium Genes for Desiccation Survival.

    Science.gov (United States)

    Mandal, Rabindra K; Kwon, Young M

    2017-01-01

    Salmonella spp., one of the most common foodborne bacterial pathogens, has the ability to survive under desiccation conditions in foods and food processing facilities for years. This raises the concerns of Salmonella infection in humans associated with low water activity foods. Salmonella responds to desiccation stress via complex pathways involving immediate physiological actions as well as coordinated genetic responses. However, the exact mechanisms of Salmonella to resist desiccation stress remain to be fully elucidated. In this study, we screened a genome-saturating transposon (Tn5) library of Salmonella Typhimurium ( S . Typhimurium) 14028s under the in vitro desiccation stress using transposon sequencing (Tn-seq). We identified 61 genes and 6 intergenic regions required to overcome desiccation stress. Salmonella desiccation resistance genes were mostly related to energy production and conversion; cell wall/membrane/envelope biogenesis; inorganic ion transport and metabolism; regulation of biological process; DNA metabolic process; ABC transporters; and two component system. More than 20% of the Salmonella desiccation resistance genes encode either putative or hypothetical proteins. Phenotypic evaluation of 12 single gene knockout mutants showed 3 mutants ( atpH, atpG , and corA ) had significantly ( p survival as compared to the wild type during desiccation survival. Thus, our study provided new insights into the molecular mechanisms utilized by Salmonella for survival against desiccation stress. The findings might be further exploited to develop effective control strategies against Salmonella contamination in low water activity foods and food processing facilities.

  1. Inv A gene specific PCR for detection of Salmonella from broilers

    Directory of Open Access Journals (Sweden)

    Thenmozhi Velayutham

    Full Text Available Poultry meat has been identified as one of the principal foodborne source of Salmonella. In this preliminary study the prevalence of Salmonella spp. contamination of broiler carcasses, were determined. Sixty samples were collected from poultry carcasses from the commercial broiler slaughtering facility in Namakkal, Tamil Nadu. The presence of Salmonella spp in collected samples was assessed by performing the pre-enrichment and enrichment culture, followed by PCR assay. The primers were selected from the invA gene specific for the detection of Salmonella spp. In this study 8.3% of poultry carcasses were found to be contaminated with Salmonella spp. In order to provide a more accurate profile of the prevalence of Salmonella spp in broiler carcasses, it is pertinent to use inv A gene specific PCR method that could be considered as an appropriate alternative to conventional culture method. [Vet. World 2011; 4(12.000: 562-564

  2. GeneQuence" Salmonella assay. Performance Tested Method 030201.

    Science.gov (United States)

    Alles, Susan; Mozola, Mark

    2009-01-01

    A study was conducted to validate the GeneQuence Salmonella DNA hybridization assay, Performance Tested Method 030201, for detection of Salmonella spp. in peanut butter. The study was organized by the AOAC Research Institute under its Emergency Response Validation program. Peanut butter samples inoculated with S. Typhimurium were prepared by an independent laboratory and shipped to study participants for testing. The set of blind-coded test samples consisted of five uninoculated controls, 20 portions inoculated with S. Typhimurium at a low level [determined by most probable number (MPN) analysis to contain 1.1 CFU/25 g portion], and 20 portions inoculated with S. Typhimurium at a higher level (11 CFU/25 g portion by MPN analysis). Samples were tested in parallel by the GeneQuence method and by the U.S. Food and Drug Administration's Bacteriological Analytical Manual reference culture procedure. All five control samples were negative by both methods. For the low-level samples, the same two samples were positive by both the GeneQuence and reference methods. For the high-level samples, the same 19 samples were positive by both methods. All positive GeneQuence assays were confirmed by plating from associated broth cultures. Agreement between the GeneQuence and reference methods was 100%. Sensitivity and specificity of the GeneQuence method were both 100%. Because neither the low- nor the high-level samples yielded the desired fractional positive results (5-15 positives out of 20 samples), a second trial was conducted. Samples in the second trial contained 0.1 and 0.5 CFU/25 g portion for the low and high levels, respectively. All five control samples were negative by both methods. For the low-level samples, the same two samples were positive by both the GeneQuence and reference methods. For the high-level samples, the same three samples were positive by both methods. All positive GeneQuence assays were confirmed by plating from associated broth cultures. Sensitivity

  3. Genomic Signature of Multidrug-Resistant Salmonella enterica Serovar Typhi Isolates Related to a Massive Outbreak in Zambia between 2010 and 2012

    DEFF Research Database (Denmark)

    Hendriksen, Rene S.; Leekitcharoenphon, Pimlapas; Lukjancenko, Oksana

    2015-01-01

    ). The isolates belonged to MLST ST1 and a new variant of the haplotype, H58B. Most isolates contained a chromosomally translocated region containing seven antimicrobial resistance genes, catA1, blaTEM-1, dfrA7, sul1, sul2, strA, and strB, and fragments of the incompatibility group Q1 (IncQ1) plasmid replicon...

  4. Live attenuated vaccines for invasive Salmonella infections

    Science.gov (United States)

    Tennant, Sharon M.; Levine, Myron M.

    2015-01-01

    Salmonella enterica serovar Typhi produces significant morbidity and mortality worldwide despite the fact that there are licensed S. 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: S. Typhi, S. Paratyphi A, S. Paratyphi B (currently uncommon but may become dominant again), S. Typhimurium, S. Enteritidis and S. 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. PMID:25902362

  5. Genome wide evolutionary analyses reveal serotype specific patterns of positive selection in selected Salmonella serotypes

    Directory of Open Access Journals (Sweden)

    Sun Qi

    2009-11-01

    Full Text Available Abstract Background The bacterium Salmonella enterica includes a diversity of serotypes that cause disease in humans and different animal species. Some Salmonella serotypes show a broad host range, some are host restricted and exclusively associated with one particular host, and some are associated with one particular host species, but able to cause disease in other host species and are thus considered "host adapted". Five Salmonella genome sequences, representing a broad host range serotype (Typhimurium, two host restricted serotypes (Typhi [two genomes] and Paratyphi and one host adapted serotype (Choleraesuis were used to identify core genome genes that show evidence for recombination and positive selection. Results Overall, 3323 orthologous genes were identified in all 5 Salmonella genomes analyzed. Use of four different methods to assess homologous recombination identified 270 genes that showed evidence for recombination with at least one of these methods (false discovery rate [FDR] ompC, a gene encoding an outer membrane protein, which has also been found to be under positive selection in other bacteria. A total of 8, 16, 7, and 5 genes showed evidence for positive selection in Choleraesuis, Typhi, Typhimurium, and Paratyphi branch analyses, respectively. Sequencing and evolutionary analyses of four genes in an additional 42 isolates representing 23 serotypes confirmed branch specific positive selection and recombination patterns. Conclusion Our data show that, among the four serotypes analyzed, (i less than 10% of Salmonella genes in the core genome show evidence for homologous recombination, (ii a number of Salmonella genes are under positive selection, including genes that appear to contribute to virulence, and (iii branch specific positive selection contributes to the evolution of host restricted Salmonella serotypes.

  6. Occurrence of integrons and antimicrobial resistance genes among Salmonella enterica from Brazil

    DEFF Research Database (Denmark)

    Peirano, G.; Agersø, Yvonne; Aarestrup, Frank Møller

    2006-01-01

    Objectives: To determine the occurrence of antimicrobial resistance genes and role of integrons among 135 anti microbial-resistant Salmonella enterica from Brazil. Methods: The presence of antimicrobial resistance genes, class 1 and 2 integrons and gene cassettes was analysed by PCR and sequencing...

  7. Regulation of Salmonella typhimurium pyr Gene Expression: Effect of Changing Both Purine and Pyrimidine Nucleotide Pools

    DEFF Research Database (Denmark)

    Jensen, Kaj Frank

    1989-01-01

    The synthesis of the pyrimidine biosynthetic enzymes is repressed by the pyrimidine nucleotide end-products of the pathway. However, purine nucleotides also play a role. In this study, I have measured expression of the pyr genes (pyrA-E) in Salmonella typhimurium strains harbouring mutations...... of each pyr gene is discussed in relation to present knowledge on gene structure and regulatory mechanism....

  8. Control of Invasive Salmonella Disease in Africa: Is There a Role for Human Challenge Models?

    OpenAIRE

    Gibani, M.M.; Jin, C.; Darton, T.C.; Pollard, A. J.

    2015-01-01

    Invasive Salmonella disease in Africa is a major public health concern. With evidence of the transcontinental spread of the Salmonella Typhi H58 haplotype, improved estimates of the burden of infection and understanding of the complex interplay of factors affecting disease transmission are needed to assist with efforts aimed at disease control. In addition to Salmonella Typhi, invasive nontyphoidal Salmonella are increasingly recognized as an important cause of febrile illness and mortality i...

  9. Identification of specific gene sequences conserved in contemporary epidemic strains of Salmonella enterica.

    Science.gov (United States)

    Kang, Min-Su; Besser, Thomas E; Hancock, Dale D; Porwollik, Steffen; McClelland, Michael; Call, Douglas R

    2006-11-01

    Genetic elements specific to recent and contemporary epidemic strains of Salmonella enterica were identified using comparative genomic analysis. Two epidemic multidrug-resistant (MDR) strains, MDR Salmonella enterica serovar Typhimurium definitive phage type 104 (DT104) and cephalosporin-resistant MDR Salmonella enterica serovar Newport, and an epidemic pansusceptible strain, Salmonella serovar Typhimurium DT160, were subjected to Salmonella gene microarray and suppression subtractive hybridization analyses. Their genome contents were compared with those of coexisting sporadic strains matched by serotype, geographic and temporal distribution, and host species origin. These paired comparisons revealed that epidemic strains of S. enterica had specific genes and gene regions that were shared by isolates of the same subtype. Most of these gene sequences are related to mobile genetic elements, including phages, plasmids, and plasmid-like and transposable elements, and some genes may encode proteins conferring growth or survival advantages. The emergence of epidemic MDR strains may therefore be associated with the presence of fitness-associated genetic factors in addition to their antimicrobial resistance genes.

  10. Quinolones in the treatment of Salmonella carriers.

    Science.gov (United States)

    Rodríguez-Noriega, E; Andrade-Villanueva, J; Amaya-Tapia, G

    1989-01-01

    Infections caused by Salmonella typhi are commonly followed by a chronic carrier state despite positive clinical and initial bacteriologic responses. The use of primary antibiotics like chloramphenicol, ampicillin, and trimethoprim-sulfamethoxazole has several major drawbacks, including in some instances the failure to prevent the carrier state. The appearance worldwide of strains with multiple resistance to the most commonly used regimens has prompted the search for new forms of therapy. Among the agents studied have been third-generation cephalosporins and quinolones, which are active in vitro against bacterial enteropathogens like S. typhi. Resolution of chronic carriage of S. typhi and other salmonellae is difficult, and regimens commonly fail (including those that combine antibiotic administration with removal of the gallbladder). In addition to being active in vitro against Salmonella species, the newer quinolones adequately penetrate the intestinal lumen, liver, bile, and gallbladder. Initial experience with norfloxacin and ciprofloxacin in oral treatment of the chronic S. typhi carrier state in adults has been promising.

  11. Molecular study on some antibiotic resistant genes in Salmonella spp. isolates

    Science.gov (United States)

    Nabi, Ari Q.

    2017-09-01

    Studying the genes related with antimicrobial resistance in Salmonella spp. is a crucial step toward a correct and faster treatment of infections caused by the pathogen. In this work Integron mediated antibiotic resistant gene IntI1 (Class I Integrase IntI1) and some plasmid mediated antibiotic resistance genes (Qnr) were scanned among the isolated non-Typhoid Salmonellae strains with known resistance to some important antimicrobial drugs using Sybr Green real time PCR. The aim of the study was to correlate the multiple antibiotics and antimicrobial resistance of Salmonella spp. with the presence of integrase (IntI1) gene and plasmid mediated quinolone resistant genes. Results revealed the presence of Class I Integrase gene in 76% of the isolates with confirmed multiple antibiotic resistances. Moreover, about 32% of the multiple antibiotic resistant serotypes showed a positive R-PCR for plasmid mediated qnrA gene encoding for nalidixic acid and ciprofloxacin resistance. No positive results could be revealed form R-PCRs targeting qnrB or qnrS. In light of these results we can conclude that the presence of at least one of the qnr genes and/or the presence of Integrase Class I gene were responsible for the multiple antibiotic resistance to for nalidixic acid and ciprofloxacin from the studied Salmonella spp. and further studies required to identify the genes related with multiple antibiotic resistance of the pathogen.

  12. Host specific differences alter the requirement for certain Salmonella genes during swine colonization.

    Science.gov (United States)

    Bearson, Bradley L; Bearson, Shawn M D

    2011-06-02

    The pathogenic potential of Salmonella is determined during the complex interaction between pathogen and host, requiring optimal regulation of multiple bacterial genetic systems within variable in vivo environments. The mouse model of systemic disease has been an extremely productive model to investigate the pathogenesis of Salmonella enterica serovar Typhimurium (S. Typhimurium). Although the mouse model is a widely used paradigm for studying the pathogenesis of systemic disease caused by Salmonella, investigations concerning food safety interventions should employ natural hosts to examine gastrointestinal colonization by Salmonella. Recent research has demonstrated specific differences in the attenuation of certain S. Typhimurium mutants in mice compared to swine. This variation in pathogenesis between the mouse model and pigs for the S. Typhimurium mutants is presumably dependent upon either the requirements for specific gene products during systemic disease (mouse) versus gastrointestinal colonization (pig) or host specific differences. In addition, host specific diversity in Salmonella colonization of swine has also been described in comparison to other food-producing animals, including cattle and chickens. Differences in Salmonella colonization and pathogenesis across diverse animal species highlight the importance of species-specific studies of gastrointestinal colonization for the development of Salmonella interventions to enhance pork safety. Copyright © 2011 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    Yang, Xiaojian; Brisbin, Jennifer; Yu, Hai; Wang, Qi; Yin, Fugui; Zhang, Yonggang; Sabour, Parviz; Sharif, Shayan; Gong, Joshua

    2014-01-01

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

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

    Science.gov (United States)

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

    2017-07-01

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

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

    OpenAIRE

    Muyyarikkandy, Muhammed Shafeekh; Amalaradjou, Mary Anne

    2017-01-01

    Salmonella Enteritidis (SE), Salmonella Typhimurium (ST), and Salmonella Heidelberg (SH) have been responsible for numerous outbreaks associated with the consumption of poultry meat and eggs. Salmonella colonization in chicken is characterized by initial attachment to the cecal epithelial cells (CEC) followed by dissemination to the liver, spleen, and oviduct. Since cecal colonization is critical to Salmonella transmission along the food chain continuum, reducing this intestinal association c...

  16. 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.

  17. Phylogenomic analysis identifies gene gains that define Salmonella enterica subspecies I.

    Science.gov (United States)

    Lienau, E Kurt; Blazar, Jeffrey M; Wang, Charles; Brown, Eric W; Stones, Robert; Musser, Steven; Allard, Marc W

    2013-01-01

    Comparative methods for analyzing whole genome sequence (WGS) data enable us to assess the genetic information available for reconstructing the evolutionary history of pathogens. We used the comparative approach to determine diagnostic genes for Salmonella enterica subspecies I. S. enterica subsp. I strains are known to infect warm-blooded organisms regularly while its close relatives tend to infect only cold-blooded organisms. We found 71 genes gained by the common ancestor of Salmonella enterica subspecies I and not subsequently lost by any member of this subspecies sequenced to date. These genes included many putative functional phenotypes. Twenty-seven of these genes are found only in Salmonella enterica subspecies I; we designed primers to test these genes for use as diagnostic sequence targets and data mined the NCBI Sequence Read Archive (SRA) database for draft genomes which carried these genes. We found that the sequence specificity and variability of these amplicons can be used to detect and discriminate among 317 different serovars and strains of Salmonella enterica subspecies I.

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

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    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.

  19. Salmonella as live trojan horse for vaccine development and cancer gene therapy.

    Science.gov (United States)

    Moreno, María; Kramer, M Gabriela; Yim, Lucía; Chabalgoity, José A

    2010-02-01

    The design of efficient vectors for vaccine development and cancer gene therapy is an area of intensive research. Bacteria-based vectors are being investigated as optimal vehicles for antigen and therapeutic gene delivery to immune and tumour cells. Attenuated Salmonella strains have shown great potential as live vectors with broad applications in human and veterinary medicine. An impressively high, and still growing, number of reports published over the last two decades have demonstrated the effectiveness in animal models of Salmonella-based therapies for the prevention and treatment of infectious and non-infectious diseases, as well as cancer. Further, the recent dramatic expansion in knowledge of genetics, biology and pathogenesis of the bacteria allows more rational design of Salmonella constructs tailored for specific applications. However, only few clinical trials have been conducted so far, and although they have conclusively demonstrated the safety of this system, the results on immunogenicity are less than optimal. Thus, more research particularly in target species is required to bring this system closer to human and veterinary use. In this review we first describe some particularities of the bacteria and its relationship with the host that could be on the basis of its success as vector, and then summarize the different strategies used so far to develop Salmonella-based vaccines for infectious diseases as well as for non-traditional indications such as prion and Alzheimer disease vaccination. Finally, we review the many different approaches that employ Salmonella for the design of new therapies for cancer.

  20. Identification of in vivo-induced genes during infection of chickens with Salmonella enterica serovar Enteritidis.

    Science.gov (United States)

    Geng, Shizhong; Liu, Zhicheng; Lin, Zhijie; Barrow, Paul; Pan, Zhiming; Li, Qiuchun; Jiao, Xinan

    2015-06-01

    Chickens are an important source of food worldwide and are often infected with food-poisoning serovars of Salmonella enterica, frequently Salmonella Enteritidis (SE), without exhibiting clinical signs of disease. Ivi (in vivo induced) genes identified using in vivo-induced antigen technology (IVIAT) are expressed only during bacterial infection and have the potential value of identifying epidemic strains and antigens which can form the basis for sub-unit vaccine development. We applied IVIAT to SE strain 50041 and identified 42 ivi genes. Eight representative ivi genes were further confirmed by qRT-PCR as being expressed only in vivo within 48 h of infection compared with that of in vitro-cultured. Although our results indicated that the identified ivi genes are expressed only in vivo, further research is needed to elucidate the exact roles of these genes during infection and pathogenesis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. In silico clustering of Salmonella global gene expression data reveals novel genes co-regulated with the SPI-1 virulence genes through HilD.

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    Martínez-Flores, Irma; Pérez-Morales, Deyanira; Sánchez-Pérez, Mishael; Paredes, Claudia C; Collado-Vides, Julio; Salgado, Heladia; Bustamante, Víctor H

    2016-11-25

    A wide variety of Salmonella enterica serovars cause intestinal and systemic infections to humans and animals. Salmonella Patogenicity Island 1 (SPI-1) is a chromosomal region containing 39 genes that have crucial virulence roles. The AraC-like transcriptional regulator HilD, encoded in SPI-1, positively controls the expression of the SPI-1 genes, as well as of several other virulence genes located outside SPI-1. In this study, we applied a clustering method to the global gene expression data of S. enterica serovar Typhimurium from the COLOMBOS database; thus genes that show an expression pattern similar to that of SPI-1 genes were selected. This analysis revealed nine novel genes that are co-expressed with SPI-1, which are located in different chromosomal regions. Expression analyses and protein-DNA interaction assays showed regulation by HilD for six of these genes: gtgE, phoH, sinR, SL1263 (lpxR) and SL4247 were regulated directly, whereas SL1896 was regulated indirectly. Interestingly, phoH is an ancestral gene conserved in most of bacteria, whereas the other genes show characteristics of genes acquired by Salmonella. A role in virulence has been previously demonstrated for gtgE, lpxR and sinR. Our results further expand the regulon of HilD and thus identify novel possible Salmonella virulence genes.

  2. Presence of Salmonella pathogenicity island 2 genes in seafood-associated Salmonella serovars and the role of the sseC gene in survival of Salmonella enterica serovar Weltevreden in epithelial cells.

    Science.gov (United States)

    Bhowmick, Patit P; Devegowda, Devananda; Ruwandeepika, H A Darshanee; Karunasagar, Iddya; Karunasagar, Indrani

    2011-01-01

    The type III secretion system encoded by the Salmonella pathogenicity island 2 (SPI-2) has a central role in the pathogenesis of systemic infections by Salmonella. Sixteen genes (ssaU, ssaB, ssaR, ssaQ, ssaO, ssaS, ssaP, ssaT, sscB, sseF, sseG, sseE, sseD, sseC, ssaD and sscA) of SPI-2 were targeted for PCR amplification in 57 seafood-associated serovars of Salmonella. The sseC gene of SPI-2 was found to be absent in two isolates of Salmonella enterica serovar Weltevreden, SW13 and SW39. Absence of sseC was confirmed by sequencing using flanking primers. SW13 had only 66 bp sequence of the sseC gene and SW39 had 58 bp sequence of this gene. A clinical isolate, S. Weltevreden--SW3, 10:r:z6--was used to construct a deletion mutant for the sseC gene. Significant reduction in the survival of SW3, 10:r:z6 ΔsseC and natural mutants SW13 and SW39 in HeLa cells suggests that sseC has a crucial role in the intracellular survival of S. Weltevreden. Expression of sseC was upregulated during the intracellular phase of both S. enterica serovar Typhimurium and clinical isolate S. Weltevreden SW3, 10:r:z6, suggesting a crucial role for this gene in the survival of S. Weltevreden inside host cells.

  3. Connexin 43 Gene Therapy Delivered by Polymer-Modified Salmonella in Murine Tumor Models

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    Wei-Kuang Wang

    2014-04-01

    Full Text Available The use of preferentially tumor-targeting bacteria as vectors is one of the most innovative approaches for the treatment of cancer. This method is based on the observation that some obligate or facultative anaerobic bacteria are capable of selectively multiplying in tumors and inhibiting their growth. Previously, we found that the tumor-targeting efficiency of Salmonella could be modulated by modifying the immune response to these bacteria by coating them with poly(allylamine hydrochloride (PAH, and these organisms are designated PAH-S.C. (S. choleraesuis. PAH can provide a useful platform for the chemical modification of Salmonella, perhaps by allowing a therapeutic gene to bind to tumor-targeting Salmonella. This study aimed to investigate the benefits of the use of PAH-S.C. for gene delivery. To evaluate this modulation, the invasion activity and gene transfer of DNA-PAH-S.C. were measured in vitro and in vivo. Treatment with PAH-S.C. carrying a tumor suppressor gene (connexin 43 resulted in inhibition of tumor growth, which suggested that tumor-targeted gene therapy using PAH-S.C. carrying a therapeutic gene could exert antitumor activities. This technique represents a promising strategy for the treatment of tumors.

  4. Stable expression of Shigella dysenteriae serotype 1 O-antigen genes integrated into the chromosome of live Salmonella oral vaccine vector Ty21a.

    Science.gov (United States)

    Dharmasena, Madushini N; Osorio, Manuel; Filipova, Svetlana; Marsh, Christina; Stibitz, Scott; Kopecko, Dennis J

    2016-09-20

    Typhoid fever and shigellosis cause high morbidity and mortality worldwide, yet no anti-Shigella vaccine is currently available. However, to protect against typhoid fever, an approved vaccine, based on the attenuated Salmonella enterica serovar Typhi strain Ty21a is available. We have investigated Ty21a as a live oral vaccine vector for expression of heterologous foreign antigens to protect against other diseases (e.g. shigellosis, anthrax, and plague). Shigella LPS is a potent vaccine antigen for serotype-specific protection against Shigellae. We previously reported the construction of a Ty21a derivative expressing S. sonnei O-antigen by insertion of a large (∼12.5 kb) operon comprising the S. sonnei O-antigen biosynthetic genes into a targeted site within the Ty21a chromosome using modified λ red recombineering methods. In the current study, S. dysenteriae 1 O-antigen biosynthetic genes from 2 separate genetic loci, rfp and rfb were assembled and inserted into the Ty21a chromosome by λ red-mediated recombineering to construct strain Ty21a-Sd. To obtain a high level of heterologous LPS expression, the native upstream promoter was replaced with the constitutive lpp promoter, which resulted in Ty21a-Sdl with enhanced heterologous LPS expression. Both Ty21a-Sd and Ty21a-Sdl elicited significant serum antibody responses in mice against both Ty21a and this heterologous Shigella LPS, and conferred protection against virulent S. dysenteriae 1 challenge. This work represents progress toward the goal of a safe and effective vaccine against Shigella. Published by Oxford University Press on behalf of FEMS 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

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

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    Xiaojian Yang

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

  6. Detection of Salmonella spp. survival and virulence in poultry feed by targeting the hilA gene.

    Science.gov (United States)

    Park, S H; Jarquin, R; Hanning, I; Almeida, G; Ricke, S C

    2011-08-01

    The objectives of this work were to evaluate immunomagnetic beads and a reverse transcriptase (RT)-PCR method for the detection of Salmonella inoculated into feed. In addition, a reverse transcriptase (RT)-PCR method was evaluated for quantifying virulence gene hilA expression of Salmonella ssp. in poultry feed matrices and utilized to determine the influence of poultry feed environmental factors on Salmonella hilA expression. An immunomagnetic separation technique was evaluated for increased recovery of Salmonella from feed. Salmonella cultures were inoculated into feed samples and exposed to heat treatments of 70°C and sampled periodically. From these samples, RNA was collected and hilA gene expression was measured relative to the housekeeping 16S rRNA gene. The immunomagnetic bead protocol increased recovery by 1 log. The up-regulation of hilA was demonstrated after 5 and 10 min of inoculated feed samples being exposed to heat treatment. From this work, the data indicate that the ability to detect live Salmonella cells in feed samples may be increased by targeting the hilA gene. Foodborne salmonellosis originating from poultry is a major problem, and feed is a leading source of contamination in poultry, but detection in feed is complicated by low concentrations. The assays and experiments in this study examine possible improvements to recovery and detection of Salmonella in feed. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  7. Type VI secretion system-associated gene clusters contribute to pathogenesis of Salmonella enterica serovar Typhimurium.

    Science.gov (United States)

    Mulder, David T; Cooper, Colin A; Coombes, Brian K

    2012-06-01

    The enteropathogen Salmonella enterica serovar Typhimurium employs a suite of tightly regulated virulence factors within the intracellular compartment of phagocytic host cells resulting in systemic dissemination in mice. A type VI secretion system (T6SS) within Salmonella pathogenicity island 6 (SPI-6) has been implicated in this process; however, the regulatory inputs and the roles of noncore genes in this system are not well understood. Here we describe four clusters of noncore T6SS genes in SPI-6 based on a comparative relationship with the T6SS-3 of Burkholderia mallei and report that the disruption of these genes results in defects in intracellular replication and systemic dissemination in mice. In addition, we show that the expression of the SPI-6-encoded Hcp and VgrG orthologs is enhanced during late stages of macrophage infection. We identify six regions that are transcriptionally active during cell infections and that have regulatory contributions from the regulators of virulence SsrB, PhoP, and SlyA. We show that levels of protein expression are very weak under in vitro conditions and that expression is not enhanced upon the deletion of ssrB, phoP, slyA, qseC, ompR, or hfq, suggesting an unknown activating factor. These data suggest that the SPI-6 T6SS has been integrated into the Salmonella Typhimurium virulence network and customized for host-pathogen interactions through the action of noncore genes.

  8. Comparative analysis of codon usage patterns and identification of predicted highly expressed genes in five Salmonella genomes

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    Mondal U

    2008-01-01

    Full Text Available Purpose: To anlyse codon usage patterns of five complete genomes of Salmonella , predict highly expressed genes, examine horizontally transferred pathogenicity-related genes to detect their presence in the strains, and scrutinize the nature of highly expressed genes to infer upon their lifestyle. Methods: Protein coding genes, ribosomal protein genes, and pathogenicity-related genes were analysed with Codon W and CAI (codon adaptation index Calculator. Results: Translational efficiency plays a role in codon usage variation in Salmonella genes. Low bias was noticed in most of the genes. GC3 (guanine cytosine at third position composition does not influence codon usage variation in the genes of these Salmonella strains. Among the cluster of orthologous groups (COGs, translation, ribosomal structure biogenesis [J], and energy production and conversion [C] contained the highest number of potentially highly expressed (PHX genes. Correspondence analysis reveals the conserved nature of the genes. Highly expressed genes were detected. Conclusions: Selection for translational efficiency is the major source of variation of codon usage in the genes of Salmonella . Evolution of pathogenicity-related genes as a unit suggests their ability to infect and exist as a pathogen. Presence of a lot of PHX genes in the information and storage-processing category of COGs indicated their lifestyle and revealed that they were not subjected to genome reduction.

  9. Vaccines against invasive Salmonella disease

    Science.gov (United States)

    MacLennan, Calman A; Martin, Laura B; Micoli, Francesca

    2014-01-01

    Though primarily enteric pathogens, Salmonellae are responsible for a considerable yet under-appreciated global burden of invasive disease. In South and South-East Asia, this manifests as enteric fever caused by serovars Typhi and Paratyphi A. In sub-Saharan Africa, a similar disease burden results from invasive nontyphoidal Salmonellae, principally serovars Typhimurium and Enteritidis. The existing Ty21a live-attenuated and Vi capsular polysaccharide vaccines target S. Typhi and are not effective in young children where the burden of invasive Salmonella disease is highest. After years of lack of investment in new Salmonella vaccines, recent times have seen increased interest in the area led by emerging-market manufacturers, global health vaccine institutes and academic partners. New glycoconjugate vaccines against S. Typhi are becoming available with similar vaccines against other invasive serovars in development. With other new vaccines under investigation, including live-attenuated, protein-based and GMMA vaccines, now is an exciting time for the Salmonella vaccine field. PMID:24804797

  10. Deteksi Kuman Salmonella Pada Ayam Goreng Yang Dijual Di Warung Makan Dan Pola Kepekaan Terhadap Berbagai Zat Antibiotika

    OpenAIRE

    Wulandari, Syuriati; Suryani, Lilis

    2008-01-01

    There are many types of bacteria that cause food poisoned. One of the bacterias is Salmonella typhi. This type of bacteria mostly inhabit the farm animals or wild animals. Mostly bacterias on cats, dogs, rats, flies and cockroaches defile chickens, meat, eggs and raw milk. Salmonella typhi defiles riped foods during storage and serving. Researches meant to detect Salmonella presence on foods sold at restaurants in Yogyakarta are rarely conducted. This research was in demand to find out the sa...

  11. Identification and characterization of novel Salmonella mobile elements involved in the dissemination of genes linked to virulence and transmission.

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    Andrea I Moreno Switt

    Full Text Available The genetic diversity represented by >2,500 different Salmonella serovars provides a yet largely uncharacterized reservoir of mobile elements that can contribute to the frequent emergence of new pathogenic strains of this important zoonotic pathogen. Currently, our understanding of Salmonella mobile elements is skewed by the fact that most studies have focused on highly virulent or common serovars. To gain a more global picture of mobile elements in Salmonella, we used prediction algorithms to screen for mobile elements in 16 sequenced Salmonella genomes representing serovars for which no prior genome scale mobile element data were available. From these results, selected mobile elements underwent further analyses in the form of validation studies, comparative analyses, and PCR-based population screens. Through this analysis we identified a novel plasmid that has two cointegrated replicons (IncI1-IncFIB; this plasmid type was found in four genomes representing different Salmonella serovars and contained a virulence gene array that had not been previously identified. A Salmonella Montevideo isolate contained an IncHI and an IncN2 plasmid, which both encoded antimicrobial resistance genes. We also identified two novel genomic islands (SGI2 and SGI3, and 42 prophages with mosaic architecture, seven of them harboring known virulence genes. Finally, we identified a novel integrative conjugative element (ICE encoding a type IVb pilus operon in three non-typhoidal Salmonella serovars. Our analyses not only identified a considerable number of mobile elements that have not been previously reported in Salmonella, but also found evidence that these elements facilitate transfer of genes that were previously thought to be limited in their distribution among Salmonella serovars. The abundance of mobile elements encoding pathogenic properties may facilitate the emergence of strains with novel combinations of pathogenic traits.

  12. Identification and characterization of novel Salmonella mobile elements involved in the dissemination of genes linked to virulence and transmission.

    Science.gov (United States)

    Moreno Switt, Andrea I; den Bakker, Henk C; Cummings, Craig A; Rodriguez-Rivera, Lorraine D; Govoni, Gregory; Raneiri, Matthew L; Degoricija, Lovorka; Brown, Stephanie; Hoelzer, Karin; Peters, Joseph E; Bolchacova, Elena; Furtado, Manohar R; Wiedmann, Martin

    2012-01-01

    The genetic diversity represented by >2,500 different Salmonella serovars provides a yet largely uncharacterized reservoir of mobile elements that can contribute to the frequent emergence of new pathogenic strains of this important zoonotic pathogen. Currently, our understanding of Salmonella mobile elements is skewed by the fact that most studies have focused on highly virulent or common serovars. To gain a more global picture of mobile elements in Salmonella, we used prediction algorithms to screen for mobile elements in 16 sequenced Salmonella genomes representing serovars for which no prior genome scale mobile element data were available. From these results, selected mobile elements underwent further analyses in the form of validation studies, comparative analyses, and PCR-based population screens. Through this analysis we identified a novel plasmid that has two cointegrated replicons (IncI1-IncFIB); this plasmid type was found in four genomes representing different Salmonella serovars and contained a virulence gene array that had not been previously identified. A Salmonella Montevideo isolate contained an IncHI and an IncN2 plasmid, which both encoded antimicrobial resistance genes. We also identified two novel genomic islands (SGI2 and SGI3), and 42 prophages with mosaic architecture, seven of them harboring known virulence genes. Finally, we identified a novel integrative conjugative element (ICE) encoding a type IVb pilus operon in three non-typhoidal Salmonella serovars. Our analyses not only identified a considerable number of mobile elements that have not been previously reported in Salmonella, but also found evidence that these elements facilitate transfer of genes that were previously thought to be limited in their distribution among Salmonella serovars. The abundance of mobile elements encoding pathogenic properties may facilitate the emergence of strains with novel combinations of pathogenic traits.

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

    Science.gov (United States)

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

    2012-12-01

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

  14. Gene Expression Profiles of Chicken Embryo Fibroblasts in Response to Salmonella Enteritidis Infection.

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    Ama Szmolka

    Full Text Available The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2 have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only 'non-immune' genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria.

  15. The Agricultural Antibiotic Carbadox Induces Phage-mediated Gene Transfer in Salmonella

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    Bradley L. Bearson

    2014-02-01

    Full Text Available Antibiotics are used for disease therapeutic or preventative effects in humans and animals, as well as for enhanced feed conversion efficiency in livestock. Antibiotics can also cause undesirable effects in microbial populations, including selection for antibiotic resistance, enhanced pathogen invasion, and stimulation of horizontal gene transfer. Carbadox is a veterinary antibiotic used in the U.S. during the starter phase of swine production for improved feed efficiency and control of swine dysentery and bacterial swine enteritis. Carbadox has been shown in vitro to induce phage-encoded Shiga toxin in Shiga toxin-producing Escherichia coli and a phage-like element transferring antibiotic resistance genes in Brachyspira hyodysenteriae, but the effect of carbadox on prophages in other bacteria is unknown. This study examined carbadox exposure on prophage induction and genetic transfer in Salmonella enterica serovar Typhimurium, a human foodborne pathogen that frequently colonizes swine without causing disease. S. Typhimurium LT2 exposed to carbadox induced prophage production, resulting in bacterial cell lysis and release of virions that were visible by electron microscopy. Carbadox induction of phage-mediated gene transfer was confirmed by monitoring the transduction of a sodCIII::neo cassette in the Fels-1 prophage from LT2 to a recipient Salmonella strain. Furthermore, carbadox frequently induced generalized transducing phages in multidrug-resistant phage type DT104 and DT120 isolates, resulting in the transfer of chromosomal and plasmid DNA that included antibiotic resistance genes. Our research indicates that exposure of Salmonella to carbadox induces prophages that can transfer virulence and antibiotic resistance genes to susceptible bacterial hosts. Carbadox-induced, phage-mediated gene transfer could serve as a contributing factor in bacterial evolution during animal production, with prophages being a reservoir for bacterial fitness

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

    Science.gov (United States)

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

    2015-08-03

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

  17. Characterization of Integrons and Resistance Genes in Salmonella Isolates from Farm Animals in Shandong Province, China

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    Xiaonan Zhao

    2017-07-01

    Full Text Available A total of 154 non-duplicate Salmonella isolates were recovered from 1,105 rectal swabs collected from three large-scale chicken farms (78/325, 24.0%, three large-scale duck farms (56/600, 9.3% and three large-scale pig farms (20/180, 11.1% between April and July 2016. Seven serotypes were identified among the 154 isolates, with the most common serotype in chickens and ducks being Salmonella enteritidis and in pigs Salmonella typhimurium. Antimicrobial susceptibility testing revealed that high antimicrobial resistance rates were observed for tetracycline (72.0% and ampicillin (69.4% in all sources. Class 1 integrons were detected in 16.9% (26/154 of these isolates and contained gene cassettes aadA2, aadA1, drfA1-aadA1, drfA12-aadA2, and drfA17-aadA5. Three β-lactamase genes were detected among the 154 isolates, and most of the isolates carried blaTEM−1(55/154, followed by blaPSE−1(14/154 and blaCTX−M−55 (11/154. Three plasmid-mediated quinolone resistance genes were detected among the 154 isolates, and most of the isolates carried qnrA (113/154, followed by qnrB (99/154 and qnrS (10/154. Fifty-four isolates carried floR among the 154 isolates. Multilocus sequence typing (MLST analysis showed that nine sequence types (STs were identified; ST11 was the most frequent genotype in chickens and ducks, and ST19 was identified in pigs. Our findings indicated that Salmonella was widespread, and the overuse of antibiotics in animals should be reduced considerably in developing countries.

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

    OpenAIRE

    Rui Figueiredo; Roderick Card; Carla Nunes; Manal AbuOun; Bagnall, Mary C.; Javier Nunez; Nuno Mendonça; Anjum, Muna F.; Gabriela Jorge da Silva

    2015-01-01

    Salmonella enterica is a zoonotic foodborne pathogen that causes acute gastroenteritis in humans. We assessed the virulence potential of one-hundred and six Salmonella strains isolated from food animals and products. A high through-put virulence genes microarray demonstrated Salmonella Pathogenicity Islands (SPI) and adherence genes were highly conserved, while prophages and virulence plasmid genes were variably present. Isolates were grouped by serotype, and virulence plasmids separated S. T...

  19. Salmonellae carrier status of food vendors in Kumasi, Ghana | Feglo ...

    African Journals Online (AJOL)

    Methods: Screening of 258 (230 females of 28 males) healthy food vendors for Salmonella typhi, and S. paratyphi A, B, and C, using stool culture, the widal test, and standard microbiological identification methods. Main outcome measures: Prevalence of chronic typhoidal Salmonellae carriers among food vendors in ...

  20. Cytotoxic mechanism of cytolethal distending toxin in nontyphoidal Salmonella serovar (Salmonella Javiana) during macrophage infection.

    Science.gov (United States)

    Williams, Katherine; Gokulan, Kuppan; Shelman, Diamond; Akiyama, Tatsuya; Khan, Ashraf; Khare, Sangeeta

    2015-02-01

    Cytolethal distending toxin B (cdtB) is a conserved virulence factor in Salmonella enterica serovar Typhi. Here we report the presence and functionality of cdtB in some nontyphoidal Salmonella (NTS) serovars, including Salmonella Javiana (cdtB+wt S. Javiana), isolated from imported food. To understand the role of cdtB in NTS serovars, a deletion mutant (cdtB(-)ΔS. Javiana) was constructed. Macrophages were infected with cdtB+wt S. Javiana (wild type), cdtB(-)Δ S. Javiana (mutant), and cdtB-negative NTS serovar (S. Typhimurium). Cytotoxic activity and transcription level of genes involved in cell death (apoptosis, autophagy, and necrosis) were assessed in infected macrophages. The cdtB+wt S. Javiana caused cellular distension as well as high degree of vacuolization and presence of the autophagosome marker LC3 in infected macrophages as compared with cdtB(-)ΔS. Javiana. The mRNA expression of genes involved in the induction of autophagy in response to toxin (Esr1 and Pik3C3) and coregulators of autophagy and apoptosis (Bax and Cyld) were significantly upregulated in cdtB(+)wt S. Javiana-infected macrophages. As autophagy destroys internalized pathogens in addition to the infected cell, it may reduce the spread of infection.

  1. A trans-acting leader RNA from a Salmonella virulence gene.

    Science.gov (United States)

    Choi, Eunna; Han, Yoontak; Cho, Yong-Joon; Nam, Daesil; Lee, Eun-Jin

    2017-09-19

    Bacteria use flagella to move toward nutrients, find its host, or retract from toxic substances. Because bacterial flagellum is one of the ligands that activate the host innate immune system, its synthesis should be tightly regulated during host infection, which is largely unknown. Here, we report that a bacterial leader mRNA from the mgtCBR virulence operon in the intracellular pathogen Salmonella enterica serovar Typhimurium binds to the fljB coding region of mRNAs in the fljBA operon encoding the FljB phase 2 flagellin, a main component of bacterial flagella and the FljA repressor for the FliC phase 1 flagellin, and degrades fljBA mRNAs in an RNase E-dependent fashion during infection. A nucleotide substitution of the fljB flagellin gene that prevents the mgtC leader RNA-mediated down-regulation increases the fljB-encoded flagellin synthesis, leading to a hypermotile phenotype inside macrophages. Moreover, the fljB nucleotide substitution renders Salmonella hypervirulent, indicating that FljB-based motility must be compromised in the phagosomal compartment where Salmonella resides. This suggests that this pathogen promotes pathogenicity by producing a virulence protein and limits locomotion by a trans-acting leader RNA from the same virulence gene during infection.

  2. Identification of a Plasmid-Mediated Quinolone Resistance Gene in Salmonella Isolates from Texas Dairy Farm Environmental Samples.

    Science.gov (United States)

    Cummings, K J; Rodriguez-Rivera, L D; Norman, K N; Ohta, N; Scott, H M

    2017-06-01

    A recent increase in plasmid-mediated quinolone resistance (PMQR) has been detected among Salmonella isolated from humans in the United States, and it is necessary to determine the sources of human infection. We had previously isolated Salmonella from dairy farm environmental samples collected in Texas, and isolates were tested for anti-microbial susceptibility. Two isolates, serotyped as Salmonella Muenster, showed the discordant pattern of nalidixic acid susceptibility and intermediate susceptibility to ciprofloxacin. For this project, whole-genome sequencing of both isolates was performed to detect genes associated with quinolone resistance. The plasmid-mediated qnrB19 gene and IncR plasmid type were identified in both isolates. To our knowledge, this is the first report of PMQR in Salmonella isolated from food animals or agricultural environments in the United States. © 2016 Blackwell Verlag GmbH.

  3. In vivo gene regulation in Salmonella spp. by a salicylate-dependent control circuit.

    Science.gov (United States)

    Royo, José Luis; Becker, Pablo Daniel; Camacho, Eva María; Cebolla, Angel; Link, Claudia; Santero, Eduardo; Guzmán, Carlos Alberto

    2007-11-01

    Systems allowing tightly regulated expression of prokaryotic genes in vivo are important for performing functional studies of bacterial genes in host-pathogen interactions and establishing bacteria-based therapies. We integrated a regulatory control circuit activated by acetyl salicylic acid (ASA) in attenuated Salmonella enterica that carries an expression module with a gene of interest under control of the XylS2-dependent Pm promoter. This resulted in 20-150-fold induction ex vivo. The regulatory circuit was also efficiently induced by ASA when the bacteria resided in eukaryotic cells, both in vitro and in vivo. To validate the circuit, we administered Salmonella spp., carrying an expression module encoding the 5-fluorocytosine-converting enzyme cytosine deaminase in the bacterial chromosome or in a plasmid, to mice with tumors. Induction with ASA before 5-fluorocytosine administration resulted in a significant reduction of tumor growth. These results demonstrate the usefulness of the regulatory control circuit to selectively switch on gene expression during bacterial infection.

  4. Survival of Salmonella Typhi and Shigella dysenteriae in dehydrated infant formula.

    Science.gov (United States)

    Day, James B; Sharma, Devang; Siddique, Nusrat; Hao, Yun-Yun D; Strain, Errol A; Blodgett, Robert J; Al-Khaldi, Sufian F

    2011-08-01

    Powdered infant formula has previously been linked to the transmission of various bacterial pathogens in infants resulting in life-threatening disease and death. Survival studies of 2 common foodborne pathogens, Salmonella enterica serovar Typhi and Shigella dysenteriae, in powdered infant formula have not been previously studied despite the potentially devastating consequences from ingestion of these organisms, particularly by newborns, in case of a natural or deliberate contamination event. Therefore, to better predict the risk of S. Typhi and S. dysenteriae infection from consumption of infant formula, the present study was undertaken to determine survival of these microorganisms in dry infant formula under varying atmospheric conditions. A 2-strain cocktail of S. Typhi and a 3-strain cocktail of S. dysenteriae were stored for up to 12 wk in dehydrated infant formula in an ambient air or nitrogen atmosphere. Viable counts of S. Typhi at 12 wk in infant formula revealed a 2.9- and 1.69-log decrease in ambient air and nitrogen atmosphere, respectively. Viable counts of S. dysenteriae at 12 wk in infant formula revealed a 0.81- and 0.42-log decrease in ambient air and nitrogen atmosphere, respectively. These results show that S. Typhi and S. dysenteriae can remain viable for prolonged periods of time in powdered infant formula, and the presence of nitrogen enhances survival. Our goal in this work was to study the survival of S. Typhi and S. dysenteriae in dehydrated storage conditions in infant formula. This interest is partially generated by the possibility of using these 2 microorganisms to deliberately contaminate the food supply. The outcome of this study will help us to have a better idea how to respond and react to the risk of deliberate food contamination. Journal of Food Science © 2011 Institute of Food Technologists® No claim to original US government works.

  5. A new chloramphenicol and florfenicol resistance gene flanked by two integron structures in Salmonella typhimurium DT104.

    Science.gov (United States)

    Arcangioli, M A; Leroy-Sétrin, S; Martel, J L; Chaslus-Dancla, E

    1999-05-15

    A new chloramphenicol resistance gene from Salmonella typhimurium DT104, designated floR, also conferring resistance to florfenicol, was characterized. Sequence analysis of the deduced FloR protein suggested that it belongs to the 12-TMS (transmembrane segments) multidrug efflux pumps family. The floR gene, and the downstream sequenced tetR and tetA tetracycline resistance genes, were surrounded by two class 1 integrons. The first one contained the resistance gene aadA2 and a deleted sulI resistance gene. The second one contained the beta-lactamase gene pse1 and a complete sulI gene. Thus, the floR gene is included in a multiresistance locus of at least 12.5 kb. Its particular organization and chromosomal location could be involved in the antibioresistance pattern stability of the DT104 Salmonella typhimurium strains.

  6. Characterisation of antimicrobial resistance in Salmonellae during 2014–2015 from four centres across India: An ICMR antimicrobial resistance surveillance network report

    Directory of Open Access Journals (Sweden)

    Sushila Dahiya

    2017-01-01

    Full Text Available Purpose: The main purpose of this study was to establish 'Antimicrobial Resistance Surveillance Network' in India and to monitor the antimicrobial susceptibility profile of clinical isolates to establish a national network across the country for monitoring antimicrobial resistance in Salmonella. Materials and Methods: This study was conducted at All India Institute of Medical Sciences, nodal centre with clinical isolates of Salmonellae collected from four centres across India, which included Christian Medical College, Vellore; Postgraduate Institute of Medical Education and Research, Chandigarh and Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry. Total 20% of the selected strains from each centre were characterised for molecular studies which included molecular mechanism of fluoroquinolones resistance and multiple locus sequence type. Results: A total of 622 Salmonellae were received from all centres during January 2014 to December 2015. Out of these 622 isolates, 380 were Salmonella Typhi, 162 were Salmonella Paratyphi A and 7 were S. Paratyphi B isolated from blood and 73 were other Salmonella serotypes. Multiple drug resistance (resistant to ampicillin, chloramphenicol and co-trimoxazole was less than 3% in S. Typhi. In S. Paratyphi A, chloramphenicol and co-trimoxazole susceptibility was 100% and 99%, respectively, whereas ampicillin susceptibility was 86% (139/161. Ciprofloxacin and nalidixic acid susceptibility was 15% (24/162 and 1% (2/162 from all centres. S. Paratyphi B was isolated from 7 patients. All isolates were third-generation cephalosporin sensitive. The most common mutations found were at codon 83 and at codon 87. We did not find any mutation in acrR gene. Efflux pump and qnr genes were not found in any isolate tested. All 86 S. Typhi isolates clustered into two sequence types - ST1 and ST2. Out of these 86 isolates, 70 S. Typhi were ST1 and 16 were ST2. All S. Paratyphi A was clustered in ST85

  7. Effect of dietary supplementation of nitrocompounds on Salmonella colonization and ileal immune gene expression in laying hens challenged with Salmonella Enteritidis.

    Science.gov (United States)

    Adhikari, Pratima; Cosby, D E; Cox, N A; Kim, W K

    2017-10-05

    Foodborne disease caused by Salmonella Enteritidis (SE) is one of the important public health and economic concerns. A study was conducted to determine the effect of supplementation with 2-nitroethanol (NE) and 2-nitropropanol (NP) on Salmonella recovery of internal organs as well as on the immune gene expression in the ileum of laying hens. Thirty-six White Leghorns were orally gavaged with nalidixic acid resistant Salmonella Enteritidis (SENR). Hens were housed individually in wire-laying cages and randomly assigned to six dietary treatments: T1 = SENR unchallenged (negative control), T2 = SENR challenged (positive control), T3 = SENR challenged + 100 ppm NE, T4 = SENR challenged + 200 ppm NE, T5 = SENR challenged + 100 ppm NP, and T6 = SENR challenged + 200 ppm NP. Hens were sampled at 7 days post inoculation (dpi). Ceca, liver with gall bladder (L/GB), and ovary samples were collected for bacteriology, and ileum samples were collected for analysis of immune gene expression. T3 and T6 significantly reduced (P GB or ovary after supplementation of either nitrocompounds. Pro- and anti-inflammatory cytokines such as interferon (IFN)-γ, interleukin (IL)-1B, IL-6, toll-like receptors (TLR)-4, and IL-10 all were significantly upregulated (P < 0.05) after SENR challenge. Supplementation at both levels of NE and NP showed a significant immune gene expression response in the ileum with reduction of IFN-γ, IL-6, TLR-4, and IL-10 mRNA expression. Overall, nitrocompounds such as NE and NP can be used in the intervention strategy to reduce Salmonella infection in hens. © 2017 Poultry Science Association Inc.

  8. O-Serotype Conversion in Salmonella Typhimurium Induces Protective Immune Responses against Invasive Non-Typhoidal Salmonella Infections

    OpenAIRE

    Li, Pei; Liu, Qing; Luo, Hongyan; Liang, Kang; Yi, Jie; Luo, Ying; Hu, Yunlong; Han, Yue; Kong, Qingke

    2017-01-01

    Salmonella infections remain a big problem worldwide, causing enteric fever by Salmonella Typhi (or Paratyphi) or self-limiting gastroenteritis by non-typhoidal Salmonella (NTS) in healthy individuals. NTS may become invasive and cause septicemia in elderly or immuno-compromised individuals, leading to high mortality and morbidity. No vaccines are currently available for preventing NTS infection in human. As these invasive NTS are restricted to several O-antigen serogroups including B1, D1, C...

  9. Application of BAX system, Tecra Unique Salmonella test, and a conventional culture method for the detection of Salmonella in ready-to-eat and raw foods.

    Science.gov (United States)

    Cheung, P-Y; Kwok, K K; Kam, K M

    2007-07-01

    To compare the BAX system, the Tecra Unique Salmonella test, and a conventional culture method for the detection of Salmonella in various foods. Ready-to-eat and raw foods were inoculated with Salmonella serotype Typhimurium, Salmonella serotype Enteritidis, Salmonella serotype Typhi, or Salmonella serotype Derby. Incubated pre-enrichment cultures were examined using the BAX system, the Tecra Unique Salmonella test, and a conventional culture method. Salmonella could be detected in all ready-to-eat food samples inoculated with S. Typhimurium, S. Enteritidis, or S. Derby, with any of the three test methods. However, false negatives were obtained with the Tecra test and the culture method when samples with higher background flora were inoculated with S. Typhi. Sensitivity test results suggested the two rapid tests performed as well as the culture method in the detection of 10(1) CFU of S. Typhimurium in 25-g cooked or raw food. The BAX system and the Tecra Unique Salmonella test demonstrated results comparable with those of the culture method in the detection of Salmonella serotypes used except S. Typhi. This is the first evaluation of the BAX system, the Tecra Unique Salmonella test, and a culture method in the detection of Salmonella in a variety of western and oriental foods.

  10. Mobilization properties of small ColE1-like plasmids carrying kanamycin resistance gene isolated from Salmonella enterica serotypes

    Science.gov (United States)

    Background: Previously we isolated and characterized various groups of small kanamycin resistance (KanR) ColE1-like plasmids from different serotypes of Salmonella enterica isolates. These plasmids all carried the aph(3)-I gene encoding the aminoglycoside phosphotransferase responsible for the kanam...

  11. Molecular profiling: Catecholamine modulation of gene expression in Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium

    Science.gov (United States)

    Investigations of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium have demonstrated that these bacterial pathogens can respond to the presence of catecholamines including norepinephrine and/or epinephrine in their environment by modulating gene expression and exhibiting various ...

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

    NARCIS (Netherlands)

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

    2000-01-01

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

  13. Epidemiology, Clinical Presentation, Laboratory Diagnosis, Antimicrobial Resistance, and Antimicrobial Management of Invasive Salmonella Infections

    Science.gov (United States)

    Sjölund-Karlsson, Maria; Gordon, Melita A.; Parry, Christopher M.

    2015-01-01

    SUMMARY Salmonella enterica infections are common causes of bloodstream infection in low-resource areas, where they may be difficult to distinguish from other febrile illnesses and may be associated with a high case fatality ratio. Microbiologic culture of blood or bone marrow remains the mainstay of laboratory diagnosis. Antimicrobial resistance has emerged in Salmonella enterica, initially to the traditional first-line drugs chloramphenicol, ampicillin, and trimethoprim-sulfamethoxazole. Decreased fluoroquinolone susceptibility and then fluoroquinolone resistance have developed in association with chromosomal mutations in the quinolone resistance-determining region of genes encoding DNA gyrase and topoisomerase IV and also by plasmid-mediated resistance mechanisms. Resistance to extended-spectrum cephalosporins has occurred more often in nontyphoidal than in typhoidal Salmonella strains. Azithromycin is effective for the management of uncomplicated typhoid fever and may serve as an alternative oral drug in areas where fluoroquinolone resistance is common. In 2013, CLSI lowered the ciprofloxacin susceptibility breakpoints to account for accumulating clinical, microbiologic, and pharmacokinetic-pharmacodynamic data suggesting that revision was needed for contemporary invasive Salmonella infections. Newly established CLSI guidelines for azithromycin and Salmonella enterica serovar Typhi were published in CLSI document M100 in 2015. PMID:26180063

  14. Global Screening of Salmonella enterica Serovar Typhimurium Genes for Desiccation Survival

    OpenAIRE

    Mandal, Rabindra K.; Kwon, Young M.

    2017-01-01

    Salmonella spp., one of the most common foodborne bacterial pathogens, has the ability to survive under desiccation conditions in foods and food processing facilities for years. This raises the concerns of Salmonella infection in humans associated with low water activity foods. Salmonella responds to desiccation stress via complex pathways involving immediate physiological actions as well as coordinated genetic responses. However, the exact mechanisms of Salmonella to resist desiccation stres...

  15. Location, location, location. Salmonella senses ethanolamine to gauge distinct host environments and coordinate gene expression

    Directory of Open Access Journals (Sweden)

    Christopher J. Anderson

    2016-01-01

    Full Text Available Chemical and nutrient signaling mediate all cellular processes, ensuring survival in response to changing environmental conditions. Ethanolamine is a component of phosphatidylethanolamine, a major phospholipid of mammalian and bacterial cell membranes. Ethanolamine is abundant in the gastrointestinal (GI tract from dietary sources as well as from the normal turnover of intestinal epithelial and bacterial cells in the gut. Additionally, mammalian cells maintain intracellular ethanolamine concentrations through low and high-affinity uptake systems and the internal recycling of phosphatidylethanolamine; therefore, ethanolamine is ubiquitous throughout the mammalian host. Although ethanolamine has profound signaling activity within mammalian cells by modulating inflammatory responses and intestinal physiology, ethanolamine is best appreciated as a nutrient for bacteria that supports growth. In our recent work (Anderson, et al. PLoS Pathog (2015, 11: e1005278, we demonstrated that Salmonella enterica serovar Typhimurium (Salmonella exploits ethanolamine signaling to adapt to distinct host environments to precisely coordinate expression of genes encoding metabolism and virulence, which ultimately enhances disease progression.

  16. Isolation of QseC-regulated genes in Salmonella enterica serovar Typhimurium by transposon mutgagenesis

    Science.gov (United States)

    Non-typhoidal Salmonella, a leading cause of U.S. foodborne disease and food-related deaths, often asymptomatically colonizes food-producing animals. In fact, >50% of U.S. swine production facilities test positive for Salmonella. The multidrug-resistant (MDR) Salmonella Typhimurium DT104 NCTC13348 c...

  17. Bayesian Modeling of MPSS Data: Gene Expression Analysis of Bovine Salmonella Infection

    KAUST Repository

    Dhavala, Soma S.

    2010-09-01

    Massively Parallel Signature Sequencing (MPSS) is a high-throughput, counting-based technology available for gene expression profiling. It produces output that is similar to Serial Analysis of Gene Expression and is ideal for building complex relational databases for gene expression. Our goal is to compare the in vivo global gene expression profiles of tissues infected with different strains of Salmonella obtained using the MPSS technology. In this article, we develop an exact ANOVA type model for this count data using a zero-inflatedPoisson distribution, different from existing methods that assume continuous densities. We adopt two Bayesian hierarchical models-one parametric and the other semiparametric with a Dirichlet process prior that has the ability to "borrow strength" across related signatures, where a signature is a specific arrangement of the nucleotides, usually 16-21 base pairs long. We utilize the discreteness of Dirichlet process prior to cluster signatures that exhibit similar differential expression profiles. Tests for differential expression are carried out using nonparametric approaches, while controlling the false discovery rate. We identify several differentially expressed genes that have important biological significance and conclude with a summary of the biological discoveries. This article has supplementary materials online. © 2010 American Statistical Association.

  18. Pleural Empyema due to Group D Salmonella

    Directory of Open Access Journals (Sweden)

    Jennifer C. Kam

    2012-01-01

    Full Text Available Non-typhi Salmonella normally presents as a bacteremia, enterocolitis, and endovascular infection but rarely manifests as pleuropulmonary disease. We present a case of a 66-year-old female with underlying pulmonary pathology, secondary to an extensive smoking history, who presented with a left-sided pleural effusion. The causative agent was identified as being group D Salmonella. Decortication of the lung was performed and the patient was discharged on antibiotics with resolution of her symptoms. This case helps to support the inclusion of Salmonella group D as a possible etiological agent of infection in the differential causes of exudative pleural effusions.

  19. Characterization and differential gene expression between two phenotypic phase variants in Salmonella enterica serovar Typhimurium.

    Directory of Open Access Journals (Sweden)

    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

  20. Prevalence of virulence and antimicrobial resistance genes in Salmonella spp. isolated from commercial chickens and human clinical isolates from South Africa and Brazil

    Directory of Open Access Journals (Sweden)

    Oliver T. Zishiri

    2016-03-01

    Full Text Available Salmonellosis is a significant public health concern around the world. The injudicious use of antimicrobial agents in poultry production for treatment, growth promotion and prophylaxis has resulted in the emergence of drug resistant strains of Salmonella. The current study was conducted to investigate the prevalence of virulence and antimicrobial resistance genes from Salmonella isolated from South African and Brazilian broiler chickens as well as human clinical isolates. Out of a total of 200 chicken samples that were collected from South Africa 102 (51% tested positive for Salmonella using the InvA gene. Of the overall 146 Salmonella positive samples that were screened for the iroB gene most of them were confirmed to be Salmonella enterica with the following prevalence rates: 85% of human clinical samples, 68.6% of South African chicken isolates and 70.8% of Brazilian chicken samples. All Salmonella isolates obtained were subjected to antimicrobial susceptibility testing with 10 antibiotics. Salmonella isolates from South African chickens exhibited resistance to almost all antimicrobial agents used, such as tetracycline (93%, trimethoprim-sulfamthoxazole (84%, trimethoprim (78.4%, kanamycin (74%, gentamicin (48%, ampicillin (47%, amoxicillin (31%, chloramphenicol (31%, erythromycin (18% and streptomycin (12%. All samples were further subjected to PCR in order to screen some common antimicrobial and virulence genes of interest namely spiC, pipD, misL, orfL, pse-1, tet A, tet B, ant (3"-la, sul 1 and sul. All Salmonella positive isolates exhibited resistance to at least one antimicrobial agent; however, antimicrobial resistance patterns demonstrated that multiple drug resistance was prevalent. The findings provide evidence that broiler chickens are colonised by pathogenic Salmonella harbouring antimicrobial resistance genes. Therefore, it is evident that there is a need for prudent use of antimicrobial agents in poultry production systems in

  1. Cloning and sequencing of hfq (host factor required for synthesis of bacteriophage Q beta RNA gene of Salmonella Typhimurium isolated from poultry

    Directory of Open Access Journals (Sweden)

    Parthasarathi Behera

    2015-05-01

    Full Text Available Aim: The aim was to clone and sequence hfq gene of Salmonella Typhimurium strain PM-45 and compare its sequence with hfq gene of other serovar of Salmonella. Materials and Methods: Salmonella Typhimurium strain PM-45 was procured from the G. B. Pant University of Agriculture and Technology, Pantnagar, India. The genomic DNA was isolated from Salmonella Typhimurium. Hfq gene was polymerase chain reaction (PCR amplified from the DNA using specific primers, which was subsequently cloned into pET32a vector and transformed into Escherichia coli BL21 pLys cells. The recombinant plasmid was isolated and subjected to restriction enzyme digestion as well as PCR. The clone was then sequenced. The sequence was analyzed and submitted in GenBank. Results: PCR produced an amplicon of 309 bp. Restriction digestion of the recombinant plasmid released the desired insert. The hfq sequence shows 100% homology with similar sequences from other Salmonella Typhimurium isolates. Both nucleotide and amino acid sequences are highly conserved. The submitted sequence is having Genbank accession no KM998764. Conclusion: Hfq, the hexameric RNA binding protein is one of the most important post-transcriptional regulator of bacteria. The sequence of hfq gene of Salmonella Typhimurium is highly conserved within and between Salmonella enterica serovars. This gene sequence is probably under heavy selection pressure to maintain the conformational integrity of its product in spite of its being not a survival gene.

  2. Biofilm formation, phenotypic production of cellulose and gene expression in Salmonella enterica decrease under anaerobic conditions.

    Science.gov (United States)

    Lamas, A; Miranda, J M; Vázquez, B; Cepeda, A; Franco, C M

    2016-12-05

    Salmonella enterica subsp. enterica is one of the main food-borne pathogens. This microorganism combines an aerobic life outside the host with an anaerobic life within the host. One of the main concerns related to S. enterica is biofilm formation and cellulose production. In this study, biofilm formation, morphotype, cellulose production and transcription of biofilm and quorum sensing-related genes of 11 S. enterica strains were tested under three different conditions: aerobiosis, microaerobiosis, and anaerobiosis. The results showed an influence of oxygen levels on biofilm production. Biofilm formation was significantly higher (PCellulose production and RDAR (red, dry, and rough) were expressed only in aerobiosis. In microaerobiosis, the strains expressed the SAW (smooth and white) morphotype, while in anaerobiosis the colonies appeared small and red. The expression of genes involved in cellulose synthesis (csgD and adrA) and quorum sensing (sdiA and luxS) was reduced in microaerobiosis and anaerobiosis in all S. enterica strains tested. This gene expression levels were less reduced in S. Typhimurium and S. Enteritidis compared to the tested serotypes. There was a relationship between the expression of biofilm and quorum sensing-related genes. Thus, the results from this study indicate that biofilm formation and cellulose production are highly influenced by atmospheric conditions. This must be taken into account as contamination with these bacteria can occur during food processing under vacuum or modified atmospheres. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Salmonella-induced changes in the gut microbiota and immune response genes transcriptome during administration of vancomycin and Bacteroides fragilis

    Directory of Open Access Journals (Sweden)

    Yu. V. Bukina

    2017-04-01

    Full Text Available The aim. To study Salmonella-induced changes in the intestinal wall microbiota, the expression of Salmonella effector proteins SipA, SopB, SopE2 and transcriptional activity of genes FFAR2, Foxp3, RORγt in rat GALT during administration of vancomycin and B.fragilis. Methods. Investigations of qualitative and quantitative composition of the microbiota of the wall of the small intestine were carried out, and the expression level of rat genes Foxp3, Rorc (Royt, FFAR2 and Salmonella effector proteins SipA, SopB and SopE2 were determined by RT-PCR, the relationship between groups of microorganisms was established. Results. Administration of B.fragilis against the background pre-treatment with vancomycin and Salmonella infection alters the quantitative composition of the microbiota in the wall of the small intestine contents: a decrease in Salmonella spp., E.coli, P.aeruginosa, Proteus spp., Enterobacter spp., Klebsiella spp. and Shigella spp., as well as increasing Bacteroides spp., E.faecalis, E.faecium and Peptostreptococcus anaerobius. The level of expression of Salmonella effector proteins in animals with the combined administration of vancomycin and S.enteritidis (I group, S.typhimurium (II group increased: SopB – 101 and 20 times; SopE2 - 80 and 2 times; SipA - 613 times (II group, and also 5-fold decrease was noted in the I group. Relative normalized number of mRNA of genes FFAR2, Foxp3, RORγt in GALT of rats in groups III and IV increased: FFAR2 - 2.7 and 5.4 times; Foxp3 - 2.5 and 85 times, RORγt level decreased by 70% and only in IV group. Conclusions. Using B.fragilis creates conditions for the correction of Salmonella-induced changes of the intestinal microbiome. Pretreatment of animals with vancomycin causes increased transcriptional activity of genes SipA, SopB and SopE2, except SipA after administration of S.enteritidis. Administration of B.fragilis increases the level of mRNA of genes FFAR2 and Foxp3 in GALT and reduces ROR

  4. Genome-wide screen for salmonella genes required for long-term systemic infection of the mouse.

    Directory of Open Access Journals (Sweden)

    2006-02-01

    Full Text Available A microarray-based negative selection screen was performed to identify Salmonella enterica serovar Typhimurium (serovar Typhimurium genes that contribute to long-term systemic infection in 129X1/SvJ (Nramp1(r mice. A high-complexity transposon-mutagenized library was used to infect mice intraperitoneally, and the selective disappearance of mutants was monitored after 7, 14, 21, and 28 d postinfection. One hundred and eighteen genes were identified to contribute to serovar Typhimurium infection of the spleens of mice by 28 d postinfection. The negatively selected mutants represent many known aspects of Salmonella physiology and pathogenesis, although the majority of the identified genes are of putative or unknown function. Approximately 30% of the negatively selected genes correspond to horizontally acquired regions such as those within Salmonella pathogenicity islands (SPI 1-5, prophages (Gifsy-1 and -2 and remnant, and the pSLT virulence plasmid. In addition, mutations in genes responsible for outer membrane structure and remodeling, such as LPS- and PhoP-regulated and fimbrial genes, were also selected against. Competitive index experiments demonstrated that the secreted SPI2 effectors SseK2 and SseJ as well as the SPI4 locus are attenuated relative to wild-type bacteria during systemic infection. Interestingly, several SPI1-encoded type III secretion system effectors/translocases are required by serovar Typhimurium to establish and, unexpectedly, to persist systemically, challenging the present description of Salmonella pathogenesis. Moreover, we observed a progressive selection against serovar Typhimurium mutants based upon the duration of the infection, suggesting that different classes of genes may be required at distinct stages of infection. Overall, these data indicate that Salmonella long-term systemic infection in the mouse requires a diverse repertoire of virulence factors. This diversity of genes presumably reflects the fact that

  5. Antimicrobial susceptibility to azithromycin among Salmonella enterica isolated from the United States

    Science.gov (United States)

    Due to emerging resistance to traditional antimicrobial agents such as ampicillin, trimethoprim-sulfamethoxazole and chloramphenicol, azithromycin is increasingly used for the treatment of invasive Salmonella infections. In the present study, 696 isolates of non-Typhi Salmonella collected from human...

  6. dfrA25, a novel trimethoprim resistance gene from Salmonella Agona isolated from a human urine sample in Brazil

    DEFF Research Database (Denmark)

    Agersø, Yvonne; Peirano, Gisele; Aarestrup, Frank Møller

    2006-01-01

    Objectives: To describe a novel trimethoprim resistance gene, designated dfrA25, which was detected as a gene cassette within a class 1 integron in Salmonella Agona. Methods: The gene was cloned into Escherichia coli MT102 and resistance to 10 different antimicrobial drugs was measured. A phyloge...... a dihydrofolate reductase of 157 amino acids with closest identity (85%) to dfrA5 dihydrofolate reductase. dfrA25 was located on a transferable plasmid (similar to 150 kb) that also harboured the tetracycline resistance gene tet(A)....

  7. Analysis of pools of targeted Salmonella deletion mutants identifies novel genes affecting fitness during competitive infection in mice.

    Directory of Open Access Journals (Sweden)

    Carlos A Santiviago

    2009-07-01

    Full Text Available Pools of mutants of minimal complexity but maximal coverage of genes of interest facilitate screening for genes under selection in a particular environment. We constructed individual deletion mutants in 1,023 Salmonella enterica serovar Typhimurium genes, including almost all genes found in Salmonella but not in related genera. All mutations were confirmed simultaneously using a novel amplification strategy to produce labeled RNA from a T7 RNA polymerase promoter, introduced during the construction of each mutant, followed by hybridization of this labeled RNA to a Typhimurium genome tiling array. To demonstrate the ability to identify fitness phenotypes using our pool of mutants, the pool was subjected to selection by intraperitoneal injection into BALB/c mice and subsequent recovery from spleens. Changes in the representation of each mutant were monitored using T7 transcripts hybridized to a novel inexpensive minimal microarray. Among the top 120 statistically significant spleen colonization phenotypes, more than 40 were mutations in genes with no previously known role in this model. Fifteen phenotypes were tested using individual mutants in competitive assays of intraperitoneal infection in mice and eleven were confirmed, including the first two examples of attenuation for sRNA mutants in Salmonella. We refer to the method as Array-based analysis of cistrons under selection (ABACUS.

  8. Comprehensive identification of Salmonella enterica serovar typhimurium genes required for infection of BALB/c mice.

    Directory of Open Access Journals (Sweden)

    Roy R Chaudhuri

    2009-07-01

    Full Text Available Genes required for infection of mice by Salmonella Typhimurium can be identified by the interrogation of random transposon mutant libraries for mutants that cannot survive in vivo. Inactivation of such genes produces attenuated S. Typhimurium strains that have potential for use as live attenuated vaccines. A quantitative screen, Transposon Mediated Differential Hybridisation (TMDH, has been developed that identifies those members of a large library of transposon mutants that are attenuated. TMDH employs custom transposons with outward-facing T7 and SP6 promoters. Fluorescently-labelled transcripts from the promoters are hybridised to whole-genome tiling microarrays, to allow the position of the transposon insertions to be determined. Comparison of microarray data from the mutant library grown in vitro (input with equivalent data produced after passage of the library through mice (output enables an attenuation score to be determined for each transposon mutant. These scores are significantly correlated with bacterial counts obtained during infection of mice using mutants with individual defined deletions of the same genes. Defined deletion mutants of several novel targets identified in the TMDH screen are effective live vaccines.

  9. multiorgan involvement due to salmonella typhi: case report

    African Journals Online (AJOL)

    the right costal margin, uniform and non tender; all other systems were normal Laboratory investigations revealed: total leukocyte count. 13 000/mm3 with a normal .... Theme: Best Practices in Paediatrics. For more information, contact: Kenya Paediatrics Association, Hurlingham Park,. 1 st Floor, Apt. C4, Off Argwings ...

  10. Pattern of multi-drug resistant Salmonella enterica serovar typhi ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-11-05

    Nov 5, 2008 ... point of sample collection was Akaraugo Hospital, Ikenegbu, Owerri,. Imo state. All the samples were packed well on ice pack and taken to the Nigeria Institute of Medical Research (NIMR), Yaba, Lagos state for isolation, antimicrobial susceptibility test and plasmid DNA isolation. Isolation. A loop full of each ...

  11. Plasmid-mediated quinolone resistance (PMQR and mutations in the topoisomerase genes of Salmonella enterica strains from Brazil

    Directory of Open Access Journals (Sweden)

    Rafaela Ferrari

    2013-01-01

    Full Text Available The objective of this study was to identify mutations in the Quinolone Resistance Determining sources Regions (QRDR of the gyrA, gyrB, parC, and parE genes and to determine if any of the qnr variants or the aac(6'-Ib-cr variant were present in strains of Salmonella spp. isolated in Brazil. A total of 126 Salmonella spp. strains from epidemic (n = 114 and poultry (n = 12 origin were evaluated. One hundred and twelve strains (88.8% were resistant to nalidixic acid (NAL and 29 (23.01% showed a reduced susceptibility to ciprofloxacin (Cip. The mutations identified were substitutions limited to the QRDR of the gyrA gene in the codons for Serine 83, Aspartate 87 and Alanine 131. The sensitivity to NAL seems to be a good phenotypic indication of distinguishing mutated and nonmutated strains in the QRDR, however the double mutation in gyrA did not cause resistance to ciprofloxacin. The qnrA1 and qnrB19 genes were detected, respectively, in one epidemic strain of S. Enteritidis and one strain of S. Corvallis of poultry origin. Despite previous detection of qnr genes in Brazil, this is the first report of qnr gene detection in Salmonella, and also the first detection of qnrB19 gene in this country. The results alert for the continuous monitoring of quinolone resistance determinants in order to minimize the emergence and selection of Salmonella spp. strains showing reduced susceptibility or resistance to quinolones.

  12. Plasmid-mediated quinolone resistance (PMQR) and mutations in the topoisomerase genes of Salmonella enterica strains from Brazil.

    Science.gov (United States)

    Ferrari, Rafaela; Galiana, Antonio; Cremades, Rosa; Rodríguez, Juan Carlos; Magnani, Marciane; Tognim, M C B; Oliveira, Tereza C R M; Royo, Gloria

    2013-01-01

    The objective of this study was to identify mutations in the Quinolone Resistance Determining sources Regions (QRDR) of the gyrA, gyrB, parC, and parE genes and to determine if any of the qnr variants or the aac(6')-Ib-cr variant were present in strains of Salmonella spp. isolated in Brazil. A total of 126 Salmonella spp. strains from epidemic (n = 114) and poultry (n = 12) origin were evaluated. One hundred and twelve strains (88.8%) were resistant to nalidixic acid (NAL) and 29 (23.01%) showed a reduced susceptibility to ciprofloxacin (Cip). The mutations identified were substitutions limited to the QRDR of the gyrA gene in the codons for Serine 83, Aspartate 87 and Alanine 131. The sensitivity to NAL seems to be a good phenotypic indication of distinguishing mutated and non-mutated strains in the QRDR, however the double mutation in gyrA did not cause resistance to ciprofloxacin. The qnrA1 and qnrB19 genes were detected, respectively, in one epidemic strain of S. Enteritidis and one strain of S. Corvallis of poultry origin. Despite previous detection of qnr genes in Brazil, this is the first report of qnr gene detection in Salmonella, and also the first detection of qnrB19 gene in this country. The results alert for the continuous monitoring of quinolone resistance determinants in order to minimize the emergence and selection of Salmonella spp. strains showing reduced susceptibility or resistance to quinolones.

  13. A mutation in the poxA gene of Salmonella enterica serovar Typhimurium alters protein production, elevates susceptibility to environmental challenges, and decreases swine colonization.

    Science.gov (United States)

    Bearson, Shawn M D; Bearson, Bradley L; Brunelle, Brian W; Sharma, Vijay K; Lee, In Soo

    2011-06-01

    Control of foodborne Salmonella within the farm-retail continuum is a complex issue since over 2500 serovars of Salmonella exist, the host range of Salmonella spp. varies greatly, and Salmonella is environmentally ubiquitous. To identify Salmonella enterica serovar Typhimurium (Salmonella Typhimurium) genes important for pathogen survival, our research group previously screened a signature-tagged mutagenesis bank in an ex vivo swine stomach content assay. A mutation in the poxA gene, a member of the gene family encoding class-II aminoacyl-tRNA synthetases, decreased survival of Salmonella Typhimurium in the ex vivo swine stomach content assay. In the current study, complementation with a plasmid-encoded poxA gene restored survival of the poxA mutant to the level of the parental, wild-type strain. In vivo analysis of the poxA mutant in the natural porcine host revealed significantly reduced fecal shedding of Salmonella, decreased colonization of the tonsils, and decreased detection of the mutant strain in the cecal contents of the pigs at 7 days postinoculation (p < 0.05). Body temperature (fever) of the pigs inoculated with wild-type Salmonella Typhimurium was significantly higher than that of pigs inoculated with the poxA mutant (p < 0.05). Two-dimensional gel electrophoresis revealed characteristic differences in the protein profile of the poxA mutant relative to the wild-type strain, indicating that deletion of poxA in Salmonella Typhimurium exerts selective effects on translation and/or posttranslational modifications of mRNA species that are necessary for stress survival and colonization of the natural swine host.

  14. Salmonella survival and differential expression of fatty acid biosynthesis-associated genes in a low-water-activity food.

    Science.gov (United States)

    Chen, W; Golden, D A; Critzer, F J

    2014-08-01

    The purpose of this study was to investigate the difference in expression of fatty acid biosynthesis genes and survival of different serotypes of Salmonella when incubated in a low-water-activity (aw ) food over a 14-day period. Stationary cells of five strains of Salmonella enterica belonging to 3 different serovars (Typhimurium ATCC 2486, Enteritidis H4267, Tennessee ARI-33, Tennessee S13952 and Tennessee K4643) were inoculated into granular sugar (aW   = 0·50) and held aerobically over a 14-day period at 25°C. Survival was determined by enumerating colonies on TSA and XLT-4 plates at 0, 1, 3, 5, 7 and 14 days. Correspondingly, gene expression was evaluated for three selected genes involved in fatty acid biosynthesis and modification (fabA, fabD and cfa). After 14 days of incubation, the population was reduced from 2·29 to 3·36 log for all five strains. Salmonella Tennessee ARI-33 and Salm. Tennessee K4643 displayed greater survival than Salm. Typhimurium and Salm. Enteritidis. The increased expression of the cfa gene (involved in cyclopropane fatty acid biosynthesis) over 14 days was found associated with strains with a lower survival rate. The fabA gene (involved in unsaturated fatty acid biosynthesis) was observed up-regulated for all strains for at least one sampling time and for Salm. Tennessee ARI-33 for all time points tested, suggesting its potential role in enhancing Salmonella survival in low aw foods. Numerous outbreaks of salmonellosis associated with low-water-activity foods have been reported. Therefore, the adaptive mechanisms utilized by Salmonella to survive in low-water-activity foods for prolonged periods of time need to be better understood. The results in this study showed that low-water-activity environments increase expression of gene fabA, which is involved in unsaturated fatty acid biosynthesis of Salmonella, while the increased expression of cfa, associated with cyclopropane fatty acid synthesis, was associated with decreased

  15. Precollaborative study of the GeneQuence Salmonella assay using 24-hour enrichment protocols for detection of Salmonella spp. in select foods.

    Science.gov (United States)

    Alles, Susan; Peng, Xuan; Wendorf, Michael; Mozola, Mark

    2007-01-01

    New enrichment protocols are described for use with a DNA hybridization (DNAH) method for detection of Salmonella spp. in select foods. GeneQuence Salmonella, in its original version, utilized a 3-stage enrichment of minimum 42 h duration. New 2-stage procedures of 24-28 h duration are described for raw poultry, raw beef, pasteurized egg products, milk chocolate, and dry pet food. In the validation study described here, a total of 345 samples were tested by the abbreviated DNAH method in parallel with either the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA/BAM) or U.S. Department of Agriculture-Food Safety and Inspection Service (USDA-FSIS) reference culture procedures. Results showed an overall sensitivity for the DNAH method of 97.1% (false-negative rate 2.9%). There were no false-positive results by the DNAH method; therefore the specificity was 100%. Overall agreement between the DNAH and reference culture methods was 98.5%. There were no significant differences in performance between the DNAH and reference methods for any of the foods tested as determined by Chi-square analysis. It is recommended that the DNAH method be subjected to AOAC collaborative study.

  16. 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.

  17. 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.

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

    DEFF Research Database (Denmark)

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

    2000-01-01

    A minitransposon mutant of Salmonella enterica serovar Typhimurium SR-11, SR-11 Fad(-), is unable to utilize gluconeogenic substrates as carbon sources and is avirulent and immunogenic when administered perorally to BALB/c mice (M. J. Utley et al., FEMS Microbiol. Lett., 163:129-134, 1998). Here......, evidence is presented that the mutation in SR-11 Fad(-) that renders the strain avirulent is in the cra gene, which encodes the Cra protein, a regulator of central carbon metabolism....

  19. Molecular basis of virulence in clinical isolates of Escherichia coli and Salmonella species from a tertiary hospital in the Eastern Cape, South Africa.

    Science.gov (United States)

    Bisi-Johnson, Mary A; Obi, Chikwelu L; Vasaikar, Sandeep D; Baba, Kamaldeen A; Hattori, Toshio

    2011-06-10

    Apart from localized gastrointestinal infections, Escherichia coli and Salmonella species are major causes of systemic disease in both humans and animals. Salmonella spp. cause invasive infections such as enteric fever, septicemia, osteomyelitis and meningitis while certain types of E. coli can cause systemic infections, includingpyelonephritis, meningitis and septicemia. These characteristic requires the involvement of a myriad of virulence factors. This study investigated the virulence factors of Escherichia coli and Salmonella species in clinical specimens from patients with diarrhoea presenting to health care centres in Oliver R. Tambo District Municipality, Eastern Cape Province, Republic of South Africa. Microbiology analysis involved the use of cultural and molecular techniques. Out of a total of 315 samples screened, Salmonella isolates were obtained in 119 (37.8%) of cases and these comprised: S. choleraesuis (6%), S. enteritidis (4%), S. eppendorf (1%), S. hadar (1%), S. isangi (8%), S. panama (1%), S. typhi (52%), S. typhimurium (25%) and untyped Salmonella spp. (2%). Among the Salmonella species 87 (73.1%) were invasive. Using molecular diagnostic methods, diarrheagenic E. coli were detected in 90 cases (28.6%): the greater proportion of this were enteroaggregative E. coli (EAEC) 37 (41.1%), enteropathogenic E. coli (EPEC) 21 (23.3%) and enterohemorrhagic E. coli (EHEC) 21 (23.3%). The predominant virulence gene among the diarrheagenic E. coli was EAEC heat-stable enterotoxin astA genes while the virulence genes identified in the Salmonella strains were 15 (12.6%) flic and 105 (88.2%) inv genes. The amino acid identity of the representative genes showed 95-100% similarity to corresponding blast searched sequence. This study showed the diversity of virulence gene expression in two major enteric pathogens. S. typhi and enteroaggregative E. coli were the predominant enteropathogens in our study area with an indication that EAEC is endemic within our study

  20. Molecular basis of virulence in clinical isolates of Escherichia coli and Salmonella species from a tertiary hospital in the Eastern Cape, South Africa

    Directory of Open Access Journals (Sweden)

    Bisi-Johnson Mary A

    2011-06-01

    Full Text Available Abstract Background Apart from localized gastrointestinal infections, Escherichia coli and Salmonella species are major causes of systemic disease in both humans and animals. Salmonella spp. cause invasive infections such as enteric fever, septicemia, osteomyelitis and meningitis while certain types of E. coli can cause systemic infections, including pyelonephritis, meningitis and septicemia. These characteristic requires the involvement of a myriad of virulence factors. Methods This study investigated the virulence factors of Escherichia coli and Salmonella species in clinical specimens from patients with diarrhoea presenting to health care centres in Oliver R. Tambo District Municipality, Eastern Cape Province, Republic of South Africa. Microbiology analysis involved the use of cultural and molecular techniques. Results Out of a total of 315 samples screened, Salmonella isolates were obtained in 119 (37.8% of cases and these comprised: S. choleraesuis (6%, S. enteritidis (4%, S. eppendorf (1%, S. hadar (1%, S. isangi (8%, S. panama (1%, S. typhi (52%, S. typhimurium (25% and untyped Salmonella spp. (2%. Among the Salmonella species 87 (73.1% were invasive. Using molecular diagnostic methods, diarrheagenic E. coli were detected in 90 cases (28.6%: the greater proportion of this were enteroaggregative E. coli (EAEC 37 (41.1%, enteropathogenic E. coli (EPEC 21 (23.3% and enterohemorrhagic E. coli (EHEC 21 (23.3%. The predominant virulence gene among the diarrheagenic E. coli was EAEC heat-stable enterotoxin astA genes while the virulence genes identified in the Salmonella strains were 15 (12.6% flic and 105 (88.2% inv genes. The amino acid identity of the representative genes showed 95-100% similarity to corresponding blast searched sequence. Conclusions This study showed the diversity of virulence gene expression in two major enteric pathogens. S. typhi and enteroaggregative E. coli were the predominant enteropathogens in our study area with an

  1. Identification of salmonella enterica serovar Kentucky genes involved in attachment to chicken skin

    Science.gov (United States)

    Background: Regardless of sanitation practices implemented to reduce Salmonella prevalence in poultry processing plants, the problem continues to be an issue. To gain an understanding of the attachment mechanism of Salmonella to broiler skin, a bioluminescent-based mutant screening assay was used. A...

  2. THE DELTA UVRB MUTATIONS IN THE AMES STRAINS OF SALMONELLA SPAN 15-119 GENES

    Science.gov (United States)

    Abstract The 4uvrB mutationesent in strains of Salmonella enterica Typhirnurium used commonly in the Salmonella (Ames) mutagenicity assay were isolated independently on separate occasions: chl-1005 (bio uvrBgal) for the hisG46-containing strains TA1535 and TA100; chl- 10...

  3. Cloning and expression of a Vi mimotope of Salmonella enterica ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-15

    Sep 15, 2009 ... A recombinant His-Vi protein of Salmonella enterica serovar Typhi was successfully constructed and cloned into an expression vector ... recombinant protein can be used to detect specific anti-Vi antibody produced by typhoid patients. Overall, the His-Vi ... E-mail: khchua@um.edu.my. Tel.:603-. 79676607.

  4. Trends of Multiple Drug Resistance in Salmonella Enterica Serovar ...

    African Journals Online (AJOL)

    Methods: This study assessed the trends in antibiotic resistance in 235 Salmonella typhi stains isolated by standard procedures from blood and/stool samples of hospitalized patients from 1997 to 2003. All the isolates were subjected to antimicrobial susceptibility testing using the following antibiotics: chloramphenicol, ...

  5. The effect of γ radiation on the expression of the virulence genes of Salmonella typhimurium and Vibrio spp.

    Science.gov (United States)

    Lim, Sangyong; Jung, Jinwoo; Kim, Dongho

    2007-11-01

    The principle benefit of food irradiation is the reduction of food-borne bacteria in food products. However, the microbiological safety with respect to increased virulence of surviving pathogens after irradiation remains an important issue with regard to the effectiveness of food irradiation. In this study, the transcriptional changes of virulence genes of Salmonella and Vibrio spp. after γ radiation were investigated by real-time PCR (RT-PCR). Samonella typhimurium is dependent upon the products of a large number of genes located within Salmonella pathogenicity islands (SPI) on the chromosome. The expressions of seven genes including four SPI genes, hilD, ssrB, pipB, and sopD, were measured at 1 h after 1 kGy irradiation. Compared with non-irradiated controls, the expression of hilD encoded within SPI1 and sopD encoding SPI1-related effector proteins was reduced about 4- and 16-fold, respectively. The expressions of Vibrio toxin genes, vvhA, ctxA, and tdh, were also monitored during the course of a growth cycle after re-inoculation of irradiated Vibrio spp. (0.5 and 1.0 kGy). The expressions of Vibrio toxin genes tested did not increase compared with non-irradiated counterparts. Results from this study indicate that γ radiation is much more likely to reduce the virulence gene expression of surviving pathogens.

  6. Development of Recombinase Polymerase Amplification Assays for Detection of Orientia tsutsugamushi or Rickettsia typhi.

    Directory of Open Access Journals (Sweden)

    Chien-Chung Chao

    Full Text Available Sensitive, specific and rapid diagnostic tests for the detection of Orientia tsutsugamushi (O. tsutsugamushi and Rickettsia typhi (R. typhi, the causative agents of scrub typhus and murine typhus, respectively, are necessary to accurately and promptly diagnose patients and ensure that they receive proper treatment. Recombinase polymerase amplification (RPA assays using a lateral flow test (RPA-nfo and real-time fluorescent detection (RPA-exo were developed targeting the 47-kDa gene of O. tsutsugamushi or 17 kDa gene of R. typhi. The RPA assay was capable of detecting O. tsutsugamushi or R. typhi at levels comparable to that of the quantitative PCR method. Both the RPA-nfo and RPA-exo methods performed similarly with regards to sensitivity when detecting the 17 kDa gene of R. typhi. On the contrary, RPA-exo performed better than RPA-nfo in detecting the 47 kDa gene of O. tsutsugamushi. The clinical performance of the O. tsutsugamushi RPA assay was evaluated using either human patient samples or infected mouse samples. Eight out of ten PCR confirmed positives were determined positive by RPA, and all PCR confirmed negative samples were negative by RPA. Similar results were obtained for R. typhi spiked patient sera. The assays were able to differentiate O. tsutsugamushi and R. typhi from other phylogenetically related bacteria as well as mouse and human DNA. Furthermore, the RPA-nfo reaction was completed in 20 minutes at 37°C followed by a 10 minute incubation at room temperature for development of an immunochromatographic strip. The RPA-exo reaction was completed in 20 minutes at 39°C. The implementation of a cross contamination proof cassette to detect the RPA-nfo fluorescent amplicons provided an alternative to regular lateral flow detection strips, which are more prone to cross contamination. The RPA assays provide a highly time-efficient, sensitive and specific alternative to other methods for diagnosing scrub typhus or murine typhus.

  7. Salmonella Enteritidis universal stress protein (usp) gene expression is stimulated by egg white and supports oviduct colonization and egg contamination in laying hens.

    Science.gov (United States)

    Raspoet, R; Gantois, I; Devloo, R; Martel, A; Haesebrouck, F; Pasmans, F; Ducatelle, R; Van Immerseel, F

    2011-11-21

    Salmonella enterica subspecies enterica serovar Enteritidis has caused a worldwide egg-associated pandemic since the mid 1980s. The exact mechanisms causing this egg tropism are still largely unknown, and only a few Salmonella genes have been implicated in the interaction with the oviduct or eggs. A in vivo expression technology screening performed previously, identified the uspA and uspB genes as being highly expressed in the chicken oviduct and in eggs. Here, we demonstrate that uspA and uspB gene expression is indeed induced after contact with egg white. Intra-oviduct inoculation of Salmonella Enteritidis uspB and uspBA mutant strains showed that the mutants had a decreased ability to colonize the magnum and isthmus of the oviduct, the organs that produce the egg white and eggshell membranes, respectively, at 7 days post-inoculation. Intravenous challenge showed that a Salmonella Enteritidis uspBA mutant strain had a decreased ability to contaminate eggs. Analogous to the function of universal stress proteins A and B in other bacterial species, we hypothesize that the Salmonella uspA and uspB genes are involved in long term persistence of Salmonella Enteritidis in harmful environments, such as in the oviduct and eggs, by conferring resistance against compounds that damage the bacterial cell membrane and DNA. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. The apeE Gene of Salmonella typhimurium Encodes an Outer Membrane Esterase Not Present in Escherichia coli

    OpenAIRE

    Carinato, Maria E.; Collin-Osdoby, Patricia; Yang, Xioming; Knox, Tina M.; Conlin, Christopher A.; Miller, Charles G.

    1998-01-01

    Salmonella typhimurium apeR mutations lead to overproduction of an outer membrane-associated N-acetyl phenylalanine β-naphthyl ester-cleaving esterase that is encoded by the apeE gene (P. Collin-Osdoby and C. G. Miller, Mol. Gen. Genet. 243:674–680, 1994). This paper reports the cloning and nucleotide sequencing of the S. typhimurium apeE gene as well as some properties of the esterase that it encodes. The predicted product of apeE is a 69.9-kDa protein which is processed to a 67-kDa species ...

  9. DNA-Based diagnostic tests for Salmonella strains targeting hilA, agfA, spvC and sef Genes

    Energy Technology Data Exchange (ETDEWEB)

    Craciunafl, C.; Keul, A. L.; Flonta, M.; Cristea, M.

    2009-07-01

    Salmoneleae are invasive enteropathogens of humans and animals. During the past decade, a dramatic increase in the occurrence of Salmonella spp infections was principally responsible for the rise of food-borne salmonellosis. The goal of this study was to evaluate the suitability of the, hilA, agfA, spvC, sef, gene amplification by PCR as a specific method for detection of Salmonella strains. (Author)

  10. Virulence gene profiling and pathogenicity characterization of non-typhoidal Salmonella accounted for invasive disease in humans.

    Directory of Open Access Journals (Sweden)

    Jotham Suez

    Full Text Available Human infection with non-typhoidal Salmonella serovars (NTS infrequently causes invasive systemic disease and bacteremia. To understand better the nature of invasive NTS (iNTS, we studied the gene content and the pathogenicity of bacteremic strains from twelve serovars (Typhimurium, Enteritidis, Choleraesuis, Dublin, Virchow, Newport, Bredeney, Heidelberg, Montevideo, Schwarzengrund, 9,12:l,v:- and Hadar. Comparative genomic hybridization using a Salmonella enterica microarray revealed a core of 3233 genes present in all of the iNTS strains, which include the Salmonella pathogenicity islands 1-5, 9, 13, 14; five fimbrial operons (bcf, csg, stb, sth, sti; three colonization factors (misL, bapA, sinH; and the invasion gene, pagN. In the iNTS variable genome, we identified 16 novel genomic islets; various NTS virulence factors; and six typhoid-associated virulence genes (tcfA, cdtB, hlyE, taiA, STY1413, STY1360, displaying a wider distribution among NTS than was previously known. Characterization of the bacteremic strains in C3H/HeN mice showed clear differences in disease manifestation. Previously unreported characterization of serovars Schwarzengrund, 9,12:l,v:-, Bredeney and Virchow in the mouse model showed low ability to elicit systemic disease, but a profound and elongated shedding of serovars Schwarzengrund and 9,12:l,v:- (as well as Enteritidis and Heidelberg due to chronic infection of the mouse. Phenotypic comparison in macrophages and epithelial cell lines demonstrated a remarkable intra-serovar variation, but also showed that S. Typhimurium bacteremic strains tend to present lower intracellular growth than gastroenteritis isolates. Collectively, our data demonstrated a common core of virulence genes, which might be required for invasive salmonellosis, but also an impressive degree of genetic and phenotypic heterogeneity, highlighting that bacteremia is a complex phenotype, which cannot be attributed merely to an enhanced invasion or

  11. Poultry Body Temperature Contributes to Invasion Control through Reduced Expression of Salmonella Pathogenicity Island 1 Genes in Salmonella enterica Serovars Typhimurium and Enteritidis

    Science.gov (United States)

    Petri, Nicholas; Daron, Caitlyn; Pereira, Rafaela; Mendoza, Mary; Hassan, Hosni M.; Koci, Matthew D.

    2015-01-01

    Salmonella enterica serovars Typhimurium (S. Typhimurium) and Enteritidis (S. Enteritidis) are foodborne pathogens, and outbreaks are often associated with poultry products. Chickens are typically asymptomatic when colonized by these serovars; however, the factors contributing to this observation are uncharacterized. Whereas symptomatic mammals have a body temperature between 37°C and 39°C, chickens have a body temperature of 41°C to 42°C. Here, in vivo experiments using chicks demonstrated that numbers of viable S. Typhimurium or S. Enteritidis bacteria within the liver and spleen organ sites were ≥4 orders of magnitude lower than those within the ceca. When similar doses of S. Typhimurium or S. Enteritidis were given to C3H/HeN mice, the ratio of the intestinal concentration to the liver/spleen concentration was 1:1. In the avian host, this suggested poor survival within these tissues or a reduced capacity to traverse the host epithelial layer and reach liver/spleen sites or both. Salmonella pathogenicity island 1 (SPI-1) promotes localization to liver/spleen tissues through invasion of the epithelial cell layer. Following in vitro growth at 42°C, SPI-1 genes sipC, invF, and hilA and the SPI-1 rtsA activator were downregulated compared to expression at 37°C. Overexpression of the hilA activators fur, fliZ, and hilD was capable of inducing hilA-lacZ at 37°C but not at 42°C despite the presence of similar levels of protein at the two temperatures. In contrast, overexpression of either hilC or rtsA was capable of inducing hilA and sipC at 42°C. These data indicate that physiological parameters of the poultry host, such as body temperature, have a role in modulating expression of virulence. PMID:26386070

  12. Typing of Typhoidal Salmonella Using Extraction of Water Soluble Whole Cell Proteins and Analysing by SDS-PAGE

    Directory of Open Access Journals (Sweden)

    R. Yousefi Mashouf

    2005-10-01

    Full Text Available Introduction & Objective : Salmonella is one of the most important genus of Enterobacteriacea family. The aim of this study was typing of typhoidal Salmonella by SDS-PAGE and comparing the results with those of serotyping method.Materials and Methods: In this study, 4 reference strains of Salmonella species, 5 reference strains of Enterobacteriacea family and 100 clinical isolates of Salmonella that were previously collected from laboratories of Hamadan medical centers were studied. Serotyping of strains were performed by Biomereux and Difco monovalent antisera. Whole-cell proteins of strains were also separated on 10% poly acrylamide gel. Gels were stained by Coomassie Brilliant Blue and analyzed by densitometry. Results: Of 100 cases of Salmonella species, 43 cases (43% were S. typhi, 20 cases (20% were S. typhymurium, 12 cases (12% were S. para typhi B, 10 cases (10% were S. para typhi C, S. para typhi A 1 case (1% and other cases were non-typhoidal Salmonella. The results of serotyping were compared with the results obtained by SDS-PAGE. Many protein bands from 220 KDa to 18.5 KDa were detected by SDS-PAGE and they were used to differentiate the strains. S. typhi serotypes were divided into 5 sub-species and S. para typhi B and C were divided each into 3 sub-species. Protein profiles of the reference strains of Salmonella were compared with protein profiles of Enterobacteriaceae species and showed some differences in major protein bands, however, they had a very similar protein band in 43 KDa area. Conclusion: Since our data was able to divide Salmonella species to sub-types and differentiate them from Enterobacteriacea species, we concluded that analsying SDS-PAGE profile of water soluble whole-cell proteins can be used for typing of these organisms and it is comparble with serotyping, nevertheless, further researches are needed to establish SDS-PAGE method and to replace it with serotyping method.

  13. Antimicrobial resistance and resistance genes in Salmonella strains isolated from broiler chickens along the slaughtering process in China.

    Science.gov (United States)

    Zhu, Yuanting; Lai, Haimei; Zou, Likou; Yin, Sheng; Wang, Chengtao; Han, Xinfeng; Xia, Xiaolong; Hu, Kaidi; He, Li; Zhou, Kang; Chen, Shujuan; Ao, Xiaolin; Liu, Shuliang

    2017-10-16

    A total of 189 Salmonella isolates were recovered from 627 samples which were collected from cecal contents of broilers, chicken carcasses, chicken meat after cutting step and frozen broiler chicken products along the slaughtering process at a slaughterhouse in Sichuan province of China. The Salmonella isolates were subjected to antimicrobial susceptibility testing to 10 categories of antimicrobial agents using the Kirby-Bauer disk diffusion method. Those antibiotics-resistant isolates were further investigated for the occurrence of resistance genes, the presence of class 1 integron as well as the associated gene cassettes, and the mutations within the gyrA and parC genes. Consequently, the prevalence of Salmonella was 30.14% (47.96% for cecal content, 18.78% for chicken carcasses, 31.33% for cutting meat and 14.00% for frozen meat, respectively). The predominant serotypes were S. Typhimurium (15.34%) and S. Enteritidis (69.84%). High resistance rates to the following drugs were observed: nalidixic acid (99.5%), ampicillin (87.8%), tetracycline (51.9%), ciprofloxacin (48.7%), trimethoprim/sulfamethoxazole (48.1%), and spectinomycin (34.4%). Antimicrobial resistance profiling showed that 60.8% of isolates were multidrug resistant (MDR), and MDR strains increased from 44.7% to 78.6% along the slaughtering line. 94.6% (n=157) of beta-lactam-resistant isolates harbored at least one resistance gene of bla TEM or bla CTX-M . The relatively low prevalence of aminoglycoside resistance genes (aac(3)-II, aac(3)-IV, and ant(2″)-I) was found in 49 (66.2%) of antibiotic-resistant isolates. The tetracycline resistance genes (tet(A), tet(B), tet(C), and tet(G) and sulfonamide resistance genes (sul1, sul2, and sul3) were identified in 84 (85.7%) and 89 (97.8%) antibiotic-resistant isolates respectively. floR was identified in 44 (97.8%) florfenicol-resistant isolates. Class 1 integron was detected in 37.4% (n=43) of the MDR isolates. Two different gene cassettes, bla OXA-30 -aad

  14. Analysis of the Salmonella regulatory network suggests involvement of SsrB and H-NS in σE-regulated SPI-2 gene expression

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    Jie eLi

    2015-02-01

    Full Text Available The extracytoplasmic functioning sigma factor σE is known to play an essential role for Salmonella enterica serovar Typhimurium to survive and proliferate in macrophages and mice. However, its regulatory network is not well characterized, especially during infection. Here we used microarray to identify genes regulated by σE in Salmonella grown in three conditions: a nutrient-rich condition and two others that mimic early and late intracellular infection. We found that in each condition σE regulated different sets of genes, and notably, several global regulators. When comparing nutrient-rich and infection-like conditions, large changes were observed in the expression of genes involved in Salmonella pathogenesis island (SPI-1 type-three secretion system (TTSS, SPI-2 TTSS, protein synthesis, and stress responses. In total, the expression of 58% of Salmonella genes was affected by σE in at least one of the three conditions. An important finding is that σE up-regulates SPI-2 genes, which are essential for Salmonella intracellular survival, by up-regulating SPI-2 activator ssrB expression at the early stage of infection and down-regulating SPI-2 repressor hns expression at a later stage. Moreover, σE is capable of countering the silencing of H-NS, releasing the expression of SPI-2 genes. This connection between E and SPI-2 genes, combined with the global regulatory effect of σE, may account for the lethality of rpoE-deficient Salmonella in murine infection.

  15. Characterization of Resistance Genes and Plasmids from Outbreaks and Illness Clusters Caused by Salmonella Resistant to Ceftriaxone in the United States, 2011-2012.

    Science.gov (United States)

    Folster, Jason P; Grass, Julian E; Bicknese, Amelia; Taylor, Julia; Friedman, Cindy R; Whichard, Jean M

    2017-03-01

    Salmonella is an important cause of foodborne illness; however, quickly identifying the source of these infections can be difficult, and source identification is a crucial step in preventing additional illnesses. Although most infections are self-limited, invasive salmonellosis may require antimicrobial treatment. Ceftriaxone, an extended-spectrum cephalosporin, is commonly used for treatment of salmonellosis. Previous studies have identified a correlation between the food animal/retail meat source of ceftriaxone-resistant Salmonella and the type of resistance gene and plasmid it carries. In this study, we examined seven outbreaks of ceftriaxone-resistant Salmonella infections, caused by serotypes Typhimurium, Newport, Heidelberg, and Infantis. All isolates were positive for a plasmid-encoded bla CMY gene. Plasmid incompatibility typing identified five IncI1 and two IncA/C plasmids. Both outbreaks containing bla CMY -IncA/C plasmids were linked to consumption of cattle products. Three of five outbreaks with bla CMY -IncI1 (ST12) plasmids were linked to a poultry source. The remaining IncI1 outbreaks were associated with ground beef (ST20) and tomatoes (ST12). In addition, we examined isolates from five unsolved clusters of ceftriaxone-resistant Salmonella infections and used our plasmid-encoded gene findings to predict the source. Overall, we identified a likely association between the source of ceftriaxone-resistant Salmonella outbreaks and the type of resistance gene/plasmid it carries.

  16. Effects of subtherapeutic concentrations of antimicrobials on gene acquisition events in Yersinia, Proteus, Shigella, and Salmonella recipient organisms in isolated ligated intestinal loops of swine.

    Science.gov (United States)

    Brewer, Matt T; Xiong, Nalee; Anderson, Kristi L; Carlson, Steve A

    2013-08-01

    To assess antimicrobial resistance and transfer of virulence genes facilitated by subtherapeutic concentrations of antimicrobials in swine intestines. 20 anesthetized pigs experimentally inoculated with donor and recipient bacteria. 4 recipient pathogenic bacteria (Salmonella enterica serotype Typhimurium, Yersinia enterocolitica, Shigella flexneri, or Proteus mirabilis) were incubated with donor bacteria in the presence of subinhibitory concentrations of 1 of 16 antimicrobials in isolated ligated intestinal loops in swine. Donor Escherichia coli contained transferrable antimicrobial resistance or virulence genes. After coincubations, intestinal contents were removed and assessed for pathogens that acquired new antimicrobial resistance or virulence genes following exposure to the subtherapeutic concentrations of antimicrobials. 3 antimicrobials (apramycin, lincomycin, and neomycin) enhanced transfer of an antimicrobial resistance plasmid from commensal E coli organisms to Yersinia and Proteus organisms, whereas 7 antimicrobials (florfenicol, hygromycin, penicillin G, roxarsone, sulfamethazine, tetracycline, and tylosin) exacerbated transfer of an integron (Salmonella genomic island 1) from Salmonella organisms to Yersinia organisms. Sulfamethazine induced the transfer of Salmonella pathogenicity island 1 from pathogenic to nonpathogenic Salmonella organisms. Six antimicrobials (bacitracin, carbadox, erythromycin, sulfathiazole, tiamulin, and virginiamycin) did not mediate any transfer events. Sulfamethazine was the only antimicrobial implicated in 2 types of transfer events. 10 of 16 antimicrobials at subinhibitory or subtherapeutic concentrations augmented specific antimicrobial resistance or transfer of virulence genes into pathogenic bacteria in isolated intestinal loops in swine. Use of subtherapeutic antimicrobials in animal feed may be associated with unwanted collateral effects.

  17. Contribution of the hdeB-like gene (SEN1493) to survival of Salmonella enterica enteritidis Nal(R) at pH 2.

    Science.gov (United States)

    Joerger, Rolf D; Choi, Steven

    2015-04-01

    Periplasmic proteins are particularly vulnerable to denaturation upon entry into a highly acid environment. In Escherichia coli, a level of protection of these proteins is afforded by acid-inducible chaperonins encoded by hdeAB. In contrast, Salmonella enterica only harbors an hdeB-like gene and it is currently not known what function it plays in this genus. In the present study, the hdeB-like gene was deleted in Salmonella enterica Enteritidis Nal(R) and Salmonella enterica Kentucky 3795. When grown overnight in tryptic soy broth (TSB) medium buffered at pH 5.5 and then exposed to TSB pH 2 for 20 min, Enteritidis wild-type strain experienced a 0.5-log10 reduction in colony-forming units, whereas the deletion strain's surviving cells were reduced by 1.6 log10. No difference in survival was observed in the corresponding Salmonella enterica Kentucky 3795 strains treated the same way. Exposure of the strains to pH 2.5 or 3 resulted in the same log reduction regardless of the presence of the hdeB-like gene. When wild-type and deletion strains of both serovars were grown in medium buffered at pH 7 prior to exposure to the acidic pHs, no difference in survival with respect to serovar or presence/absence of the hdeB-like gene was found. Salmonella enterica Enteritidis Nal(R) carrying its own or the intragenic region upstream of the hdeB-like from Salmonella enterica Kentucky 3795 cloned in front of the gfp gene from pFPV25 showed maximum fluorescence when grown at pH 5.5, whereas the corresponding plasmid-carrying Salmonella enterica Kentucky strains did not exhibit fluorescence regardless of the pH of the growth medium. Therefore, the hdeB-like gene in Salmonella enterica Enteritidis, but not in Salmonella enterica Kentucky 3795, contributed to survival at pH 2 and its expression is responsive to the pH of the medium.

  18. LeuO is a global regulator of gene expression in Salmonella enterica serovar Typhimurium

    DEFF Research Database (Denmark)

    Dillon, Shane C.; Espinosa, Elena; Hokamp, Karsten

    2012-01-01

    We report the first investigation of the binding of the Salmonella enterica LeuO LysR‐type transcription regulator to its genomic targets in vivo. Chromatin‐immunoprecipitation‐on‐chip identified 178 LeuO binding sites on the chromosome of S. enterica serovar Typhimurium strain SL1344. These site...

  19. Gene expression response of the rat small intestine following oral salmonella infection

    NARCIS (Netherlands)

    Rodenburg, G.C.H.; Bovee-Oudenhoven, I.M.J.; Kramer, E.H.M.; Meer, van der R.; Keijer, J.

    2007-01-01

    Data on the molecular response of the intestine to the food-borne pathogen Salmonella are derived from in vitro studies, whereas in vivo data are lacking. We performed an oral S. enteritidis infection study in Wistar rats to obtain insight in the in vivo response in time. Expression profiles of

  20. Characterization of Salmonella enterica and detection of the virulence genes specific to diarrheagenic Escherichia coli from poultry carcasses in Ouagadougou, Burkina Faso.

    Science.gov (United States)

    Kagambèga, Assèta; Barro, Nicolas; Traoré, Alfred S; Siitonen, Anja; Haukka, Kaisa

    2012-07-01

    One hundred chicken carcasses purchased from three markets selling poultry in Ouagadougou, Burkina Faso, between June 2010 and October 2010 were examined for their microbiological quality. The presence of Salmonella was investigated using standard bacteriological procedures, and the isolates obtained were serotyped and tested for antimicrobial susceptibility. The presence of virulence-associated genes of the five main pathogroups of diarrheagenic Escherichia coli-Shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), enterotoxigenic E. coli, and enteroinvasive E. coli-was investigated using 16-plex polymerase chain reaction (PCR) on the mixed bacterial cultures from the poultry samples. Of the 100 chicken carcasses studied, 57 were contaminated by Salmonella; 16 different serotypes were identified, the most frequent being Salmonella Derby, found in 28 samples. Four Salmonella strains were resistant to tetracycline, and two were resistant to streptomycin. Based on the PCR detection of the virulence genes, in total, 45 carcasses were contaminated by three pathogroups of E. coli: STEC, EPEC, or EAEC. The STEC and EPEC virulence genes were detected on six and 39 carcasses, respectively. EAEC virulence genes were only detected in combination with those of EPEC (on 11 carcasses) or STEC (on two carcasses). The STEC-positive carcasses contained the genes stx(1), stx(2), eaeA, escV, and ent in different combinations. None of the EPEC-positive carcasses contained the bfp gene, indicating that only atypical EPEC was present. EAEC virulence genes detected were aggR and/or pic. The high proportion of chicken carcasses contaminated by Salmonella and diarrheagenic E. coli indicates a potential food safety risk for consumers and highlights the necessity of public awareness of these pathogens.

  1. Antibiogram pattern of Salmonella in blood samples of enteric fever patients at Lalitpur, Nepal

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    Biraj Gurung

    2017-01-01

    Full Text Available Objective: To determine the status of isolation blood stream serotypes of enteric fever pathogens and their antibiotic susceptibility patterns and to guide clinicians for appropriate therapy. Methods: Samples were examined by microbiological techniques to identify the causative agent and determine their antimicrobial susceptibility patterns by Kirby-Bauer disk diffusion methods and interpreted as per Clinical and Laboratory Standards Institute guidelines. Results: Among 403 blood samples, 76 (18.85% showed growth for Salmonella isolates. Distribution of Salmonella typhi and Salmonella paratyphi A isolates were found to be 54% and 46% respectively. Among 76 Salmonella isolates, 28 (36.84% were from male and remaining 48 (63.15% were from female belonging to all age-groups. Multidrug-resistance was found to be 17% among the Salmonella isolates. Nalidixic acid resistance was 73.68% in Salmonella with higher proportion in Salmonella paratyphi A (85.7% in comparison to Salmonella typhi (63.42%. Salmonella isolates demonstrated 100% susceptibility to azithromycin, ceftriazone, ciprofloxacin, ofloxacin and imipenem. Conclusions: The need of continual surveillance of resistance levels to guide clinicians for appropriate therapy based on the antibiotic susceptibility pattern for Salmonella isolates is sustained with discouragement in misuse of antibiotics prior to prescription as multidrugresistance-nalidixic acid resistant strains.

  2. Nucleotide polymorphism and evolution in the glyceraldehyde-3-phosphate dehydrogenase gene (gapA) in natural populations of Salmonella and Escherichia coli.

    Science.gov (United States)

    Nelson, K; Whittam, T S; Selander, R K

    1991-08-01

    Nucleotide sequences of the gapA gene, encoding the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase, were determined for 16 strains of Salmonella and 13 strains of Escherichia coli recovered from natural populations. Pairs of sequences from strains representing the eight serovar groups of Salmonella differed, on average, at 3.8% of nucleotide sites and 1.1% of inferred amino acids, and comparable values for E. coli were an order of magnitude smaller (0.2% and 0.1%, respectively). The rate of substitution at synonymous sites was significantly higher for codons specifying the catalytic domain of the enzyme than for those encoding the NAD(+)-binding domain, but the nonsynonymous substitution rate showed the opposite relationship. For Salmonella, statistical tests for nonrandom clustering of polymorphic sites failed to provide evidence that intragenic recombination or gene conversion has contributed to the generation of allelic diversity. The topology of a tree constructed from the gapA sequences was generally similar to that of phylogenetic trees of the strains based on multilocus enzyme electrophoresis, but the level of divergence of gapA in Salmonella group V from other Salmonella and E. coli strains is much greater than that indicated by DNA hybridization for the genome as a whole.

  3. Unstable tandem gene amplification generates heteroresistance (variation in resistance within a population) to colistin in Salmonella enterica.

    Science.gov (United States)

    Hjort, Karin; Nicoloff, Hervé; Andersson, Dan I

    2016-10-01

    Heteroresistance, a phenomenon where subpopulations of a bacterial isolate exhibit different susceptibilities to an antibiotic, is a growing clinical problem where the underlying genetic mechanisms in most cases remain unknown. We isolated colistin resistant mutants in Escherichia coli and Salmonella enterica serovar Typhimurium at different concentrations of colistin. Genetic analysis showed that genetically stable pmrAB point mutations were responsible for colistin resistance during selection at high drug concentrations for both species and at low concentrations for E. coli. In contrast, for S. Typhimurium mutants selected at low colistin concentrations, amplification of different large chromosomal regions conferred a heteroresistant phenotype. All amplifications included the pmrD gene, which encodes a positive regulator that up-regulates proteins that modify lipid A, and as a result increase colistin resistance. Inactivation and over-expression of the pmrD gene prevented and conferred resistance, respectively, demonstrating that the PmrD protein is required and sufficient to confer resistance. The heteroresistance phenotype is explained by the variable gene dosage of pmrD in a population, where sub-populations with different copy number of the pmrD gene show different levels of colistin resistance. We propose that variability in gene copy number of resistance genes can explain the heteroresistance observed in clinically isolated pathogenic bacteria. © 2016 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd.

  4. The Salmonella enterica PhoP directly activates the horizontally acquired SPI-2 gene sseL and is functionally different from a S. bongori ortholog.

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    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.

  5. Analysis of Salmonella enterica serotype paratyphi A gene expression in the blood of bacteremic patients in Bangladesh.

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    Alaullah Sheikh

    2010-12-01

    Full Text Available Salmonella enterica serotype Paratyphi A is a human-restricted cause of paratyphoid fever, accounting for up to a fifth of all cases of enteric fever in Asia.In this work, we applied an RNA analysis method, Selective Capture of Transcribed Sequences (SCOTS, and cDNA hybridization-microarray technology to identify S. Paratyphi A transcripts expressed by bacteria in the blood of three patients in Bangladesh. In total, we detected 1,798 S. Paratyphi A mRNAs expressed in the blood of infected humans (43.9% of the ORFeome. Of these, we identified 868 in at least two patients, and 315 in all three patients. S. Paratyphi A transcripts identified in at least two patients encode proteins involved in energy metabolism, nutrient and iron acquisition, vitamin biosynthesis, stress responses, oxidative stress resistance, and pathogenesis. A number of detected transcripts are expressed from PhoP and SlyA-regulated genes associated with intra-macrophage survival, genes contained within Salmonella Pathogenicity Islands (SPIs 1-4, 6, 10, 13, and 16, as well as RpoS-regulated genes. The largest category of identified transcripts is that of encoding proteins with unknown function. When comparing levels of bacterial mRNA using in vivo samples collected from infected patients to samples from in vitro grown organisms, we found significant differences for 347, 391, and 456 S. Paratyphi A transcripts in each of three individual patients (approximately 9.7% of the ORFeome. Of these, expression of 194 transcripts (4.7% of ORFs was concordant in two or more patients, and 41 in all patients. Genes encoding these transcripts are contained within SPI-1, 3, 6 and 10, PhoP-regulated genes, involved in energy metabolism, nutrient acquisition, drug resistance, or uncharacterized genes. Using quantitative RT-PCR, we confirmed increased gene expression in vivo for a subset of these genes.To our knowledge, we describe the first microarray-based transcriptional analysis of a pathogen

  6. Host response transcriptional profiling reveals extracellular components and ABC (ATP-binding cassette transporters gene enrichment in typhoid fever-infected Nigerian children

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    Resau James H

    2011-09-01

    Full Text Available Abstract Background Salmonella enterica serovar Typhi (S. Typhi is a human-specific pathogen that causes typhoid fever, and remains a global health problem especially in developing countries. Its pathogenesis is complex and host response is poorly understood. In Africa, typhoid fever can be a major cause of morbidity in young infected children. The onset of the illness is insidious and clinical diagnosis is often unreliable. Gold standard blood culture diagnostic services are limited, thus rapid, sensitive, and affordable diagnostic test is essential in poor-resourced clinical settings. Routine typhoid fever vaccination is highly recommended but currently licensed vaccines provide only 55-75% protection. Recent epidemiological studies also show the rapid emergence of multi-drug resistant S. Typhi strains. High-throughput molecular technologies, such as microarrays, can dissect the molecular mechanisms of host responses which are S. Typhi-specific to provide a comprehensive genomic component of immunological responses and suggest new insights for diagnosis and treatment. Methods Global transcriptional profiles of S. Typhi-infected young Nigerian children were obtained from their peripheral blood and compared with that of other bacteremic infections using Agilent gene expression microarrays. The host-response profiles of the same patients in acute vs. convalescent phases were also determined. The top 96-100 differentially-expressed genes were identified and four genes were validated by quantitative real-time PCR. Gene clusters were obtained and functional pathways were predicted by DAVID (Database for Annotation, Visualization and Integrated Discovery. Results Transcriptional profiles from S. Typhi-infected children could be distinguished from those of other bacteremic infections. Enriched gene clusters included genes associated with extracellular peptides/components such as lipocalin (LCN2 and systemic immune response which is atypical in

  7. Cellulitis Due to Salmonella infantis.

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    Satish R Patil

    2013-01-01

    Full Text Available Bacteria of the genus Salmonella are highly adapted for the growth in both humans and animals and cause a wide spectrum of disease. The growth of Serotypes S. typhi and S. paratyphi is restricted to human hosts, in whom these organisms cause enteric (typhoid fever. The remaining Serotypes (non typhoidal Salmonella or NTS can colonize the gastrointestinal tracts of the broad range of animals, including mammals, reptiles, birds and insects. The usual clinical presentation of non-typhoidal salmonellae (NTS infection is self limited gastroenteritis; however bacteremia and focal extra intestinal infection may occur. However salmonella localization to the skin presenting as cutaneous ulceration is regarded as a rare event. Rates of morbidity and mortality associated with NTS are highest among the elderly, infants, and immunocompromised individuals, including those with hemoglobinopathies, HIV infection, or infections that cause blockade of the reticuloendothelial system. We isolated S.infantis in 50 years old man with left leg cellulitis. The serotype was confirmed at Central Research Institute, Kasauli.

  8. Identification of the crucial genes in the elimination and survival process of Salmonella enterica ser. Pullorum in the chicken spleen.

    Science.gov (United States)

    Ma, T; Xu, L; Wang, H; Guo, X; Li, Z; Wan, F; Chen, J; Liu, L; Liu, X; Chang, G; Chen, G

    2017-06-01

    Salmonella enterica ser. Pullorum is one of the most easily re-infecting pathogens in poultry production because of its mechanism of escaping from immune elimination. We used the transcriptome method to investigate the variation in gene expression in chicken spleen resulting from the interaction between hosts and S. Pullorum in the survival process. The expression of various genes related to the maturation and activation of B cells was activated before S. Pullorum was eliminated, which might help S. Pullorum escape from the elimination process. The suppression of some genes involved in the fusion of autophagosomes and lysosomes, such as MYO6, was identified and may be regulated by the secretion systems of S. Pullorum. In addition, a large proportion of these differentially expressed genes could be localized in the identified quantitative trait loci regions associated with the antibody response to bacteria. Collectively, these identified genes provided an outline for further understanding the interaction between chicken immune cells and S. Pullorum in chicken spleen. © 2017 Stichting International Foundation for Animal Genetics.

  9. Bistable expression of virulence genes in salmonella leads to the formation of an antibiotic-tolerant subpopulation.

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    Markus Arnoldini

    2014-08-01

    Full Text Available Phenotypic heterogeneity can confer clonal groups of organisms with new functionality. A paradigmatic example is the bistable expression of virulence genes in Salmonella typhimurium, which leads to phenotypically virulent and phenotypically avirulent subpopulations. The two subpopulations have been shown to divide labor during S. typhimurium infections. Here, we show that heterogeneous virulence gene expression in this organism also promotes survival against exposure to antibiotics through a bet-hedging mechanism. Using microfluidic devices in combination with fluorescence time-lapse microscopy and quantitative image analysis, we analyzed the expression of virulence genes at the single cell level and related it to survival when exposed to antibiotics. We found that, across different types of antibiotics and under concentrations that are clinically relevant, the subpopulation of bacterial cells that express virulence genes shows increased survival after exposure to antibiotics. Intriguingly, there is an interplay between the two consequences of phenotypic heterogeneity. The bet-hedging effect that arises through heterogeneity in virulence gene expression can protect clonal populations against avirulent mutants that exploit and subvert the division of labor within these populations. We conclude that bet-hedging and the division of labor can arise through variation in a single trait and interact with each other. This reveals a new degree of functional complexity of phenotypic heterogeneity. In addition, our results suggest a general principle of how pathogens can evade antibiotics: Expression of virulence factors often entails metabolic costs and the resulting growth retardation could generally increase tolerance against antibiotics and thus compromise treatment.

  10. High-level fluoroquinolone resistant Salmonella enterica serovar Kentucky ST198 epidemic clone with IncA/C conjugative plasmid carrying bla(CTX-M-25) gene.

    Science.gov (United States)

    Wasyl, Dariusz; Kern-Zdanowicz, Izabela; Domańska-Blicharz, Katarzyna; Zając, Magdalena; Hoszowski, Andrzej

    2015-01-30

    Multidrug resistant Salmonella Kentucky strains have been isolated from turkeys in Poland since 2009. Multiple mutations within chromosomal genes gyrA and parC were responsible for high-level ciprofloxacin resistance. One of the isolates was extended spectrum β-lactamase- (ESBL) positive: the strain 1643/2010 carried a conjugative 167,779 bps plasmid of IncA/C family. The sequence analysis revealed that it carried a blaCTX-M-25 gene and an integron with another β-lactamase encoding gene-blaOXA-21. This is the first known report of a CTX-M-25 encoding gene both in Poland and in Salmonella Kentucky world-wide, as well as in the IncA/C plasmid. Analysis of the integron showed a novel arrangement of gene cassettes-aacA4, aacC-A1 and blaOXA-21 where the latter might result from an intergeneric gene transfer. The study confirmed Salmonella Kentucky population isolated in Poland belongs to global epidemics of high level fluoroquinolone resistant clone ST198 that can carry rare β-lactamase genes. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Lysogeny and Bacteriocinogeny in Salmonella, Shigella, Bacillus Pyocyaneus and Neisseria meningitidis.

    Science.gov (United States)

    1979-09-01

    Ibrahim Mohamed Shabana "Flora of acopharynx in Allergic and Bacterial Rhinitis with Special Stress on Neisseria Spesies". 5. J . Blham Metuwie uSerology...almunelli typhi-murii Related to Txvo- Uutbretaks of Food Poisgoning in NE.Ypt.’ JLl-JLzhar Miedical JuuruEJ "Iln Press). Phag Patterns of Salmonella

  12. Salmonella Pathogenicity and Host Adaptation in Chicken-Associated Serovars

    Science.gov (United States)

    Johnson, Timothy J.; Ricke, Steven C.; Nayak, Rajesh; Danzeisen, Jessica

    2013-01-01

    SUMMARY Enteric pathogens such as Salmonella enterica cause significant morbidity and mortality. S. enterica serovars are a diverse group of pathogens that have evolved to survive in a wide range of environments and across multiple hosts. S. enterica serovars such as S. Typhi, S. Dublin, and S. Gallinarum have a restricted host range, in which they are typically associated with one or a few host species, while S. Enteritidis and S. Typhimurium have broad host ranges. This review examines how S. enterica has evolved through adaptation to different host environments, especially as related to the chicken host, and continues to be an important human pathogen. Several factors impact host range, and these include the acquisition of genes via horizontal gene transfer with plasmids, transposons, and phages, which can potentially expand host range, and the loss of genes or their function, which would reduce the range of hosts that the organism can infect. S. Gallinarum, with a limited host range, has a large number of pseudogenes in its genome compared to broader-host-range serovars. S. enterica serovars such as S. Kentucky and S. Heidelberg also often have plasmids that may help them colonize poultry more efficiently. The ability to colonize different hosts also involves interactions with the host's immune system and commensal organisms that are present. Thus, the factors that impact the ability of Salmonella to colonize a particular host species, such as chickens, are complex and multifactorial, involving the host, the pathogen, and extrinsic pressures. It is the interplay of these factors which leads to the differences in host ranges that we observe today. PMID:24296573

  13. Identification of Salmonella enterica genes with a role in persistence on lettuce leaves during cold storage by recombinase-based in vivo expression technology.

    Science.gov (United States)

    Kroupitski, Y; Brandl, M T; Pinto, R; Belausov, E; Tamir-Ariel, D; Burdman, S; Sela Saldinger, S

    2013-04-01

    Recurrent outbreaks of enteric illness linked to lettuce and a lack of efficacious strategies to decontaminate produce underscores the need for a better understanding of the molecular interactions of foodborne pathogens with plants. This study aimed at identifying Salmonella enterica genes involved in the persistence of this organism on post-harvest lettuce during cold storage using recombinase-based in vivo expression technology (RIVET). In total, 37 potentially induced loci were identified in four distinct screenings. Knockout mutations in eight upregulated genes revealed that four of them have a role in persistence of the pathogen in this system. These genes included stfC, bcsA, misL, and yidR, encoding a fimbrial outer membrane usher, a cellulose synthase catalytic subunit, an adhesin of the autotransporter family expressed from the Salmonella pathogenicity island-3, and a putative ATP/GTP-binding protein, respectively. bcsA, misL, and yidR but not stfC mutants were impaired also in attachment and biofilm formation, suggesting that these functions are required for survival of S. enterica on post-harvest lettuce. This is the first report that MisL, which has a role in Salmonella binding to fibronectin in animal hosts, is involved also in adhesion to plant tissue. Hence, our study uncovered a new plant attachment factor in Salmonella and demonstrates that RIVET is an effective approach for investigating human pathogen-plant interactions in a post-harvest leafy vegetable.

  14. 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.

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

    Directory of Open Access Journals (Sweden)

    Ramachandran Vinoy K

    2012-01-01

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

  16. Three Epidemics of Invasive Multidrug-Resistant Salmonella Bloodstream Infection in Blantyre, Malawi, 1998–2014

    Science.gov (United States)

    Feasey, Nicholas A.; Masesa, Clemens; Jassi, Chikondi; Faragher, E. Brian; Mallewa, Jane; Mallewa, Macpherson; MacLennan, Calman A.; Msefula, Chisomo; Heyderman, Robert S.; Gordon, Melita A.

    2015-01-01

    Background. The Malawi Liverpool Wellcome Trust Clinical Research Programme (MLW) has routinely collected specimens for blood culture from febrile patients, and cerebrospinal fluid from patients with suspected meningitis, presenting to Queen Elizabeth Central Hospital (QECH), Blantyre, Malawi, since 1998. Methods. We present bloodstream infection (BSI) and meningitis surveillance data from 1998 to 2014. Automated blood culture, manual speciation, serotyping, and antimicrobial susceptibility testing were performed at MLW. Population data for minimum-incidence estimates in urban Blantyre were drawn from published estimates. Results. Between 1998 and 2014, 167 028 blood cultures were taken from adult and pediatric medical patients presenting to QECH; Salmonella Typhi was isolated on 2054 occasions (1.2%) and nontyphoidal Salmonella (NTS) serovars were isolated 10 139 times (6.1%), of which 8017 (79.1%) were Salmonella Typhimurium and 1608 (15.8%) were Salmonella Enteritidis. There were 392 cases of NTS meningitis and 9 cases of Salmonella Typhi meningitis. There have been 3 epidemics of Salmonella BSI in Blantyre; Salmonella Enteritidis from 1999 to 2002, Salmonella Typhimurium from 2002 to 2008, and Salmonella Typhi, which began in 2011 and was ongoing in 2014. Multidrug resistance has emerged in all 3 serovars and is seen in the overwhelming majority of isolates, while resistance to third-generation cephalosporins and fluoroquinolones is currently uncommon but has been identified. Conclusions. Invasive Salmonella disease in Malawi is dynamic and not clearly attributable to a single risk factor, although all 3 epidemics were associated with multidrug resistance. To inform nonvaccine and vaccine interventions, reservoirs of disease and modes of transmission require further investigation. PMID:26449953

  17. Three Epidemics of Invasive Multidrug-Resistant Salmonella Bloodstream Infection in Blantyre, Malawi, 1998-2014.

    Science.gov (United States)

    Feasey, Nicholas A; Masesa, Clemens; Jassi, Chikondi; Faragher, E Brian; Mallewa, Jane; Mallewa, Macpherson; MacLennan, Calman A; Msefula, Chisomo; Heyderman, Robert S; Gordon, Melita A

    2015-11-01

    The Malawi Liverpool Wellcome Trust Clinical Research Programme (MLW) has routinely collected specimens for blood culture from febrile patients, and cerebrospinal fluid from patients with suspected meningitis, presenting to Queen Elizabeth Central Hospital (QECH), Blantyre, Malawi, since 1998. We present bloodstream infection (BSI) and meningitis surveillance data from 1998 to 2014. Automated blood culture, manual speciation, serotyping, and antimicrobial susceptibility testing were performed at MLW. Population data for minimum-incidence estimates in urban Blantyre were drawn from published estimates. Between 1998 and 2014, 167,028 blood cultures were taken from adult and pediatric medical patients presenting to QECH; Salmonella Typhi was isolated on 2054 occasions (1.2%) and nontyphoidal Salmonella (NTS) serovars were isolated 10,139 times (6.1%), of which 8017 (79.1%) were Salmonella Typhimurium and 1608 (15.8%) were Salmonella Enteritidis. There were 392 cases of NTS meningitis and 9 cases of Salmonella Typhi meningitis. There have been 3 epidemics of Salmonella BSI in Blantyre; Salmonella Enteritidis from 1999 to 2002, Salmonella Typhimurium from 2002 to 2008, and Salmonella Typhi, which began in 2011 and was ongoing in 2014. Multidrug resistance has emerged in all 3 serovars and is seen in the overwhelming majority of isolates, while resistance to third-generation cephalosporins and fluoroquinolones is currently uncommon but has been identified. Invasive Salmonella disease in Malawi is dynamic and not clearly attributable to a single risk factor, although all 3 epidemics were associated with multidrug resistance. To inform nonvaccine and vaccine interventions, reservoirs of disease and modes of transmission require further investigation. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.

  18. Salmonella Osteomyelitis

    National Research Council Canada - National Science Library

    McAnearney, S; McCall, D

    2015-01-01

    .... Salmonella as an aetiological agent in osteomyelitis is essentially rare and salmonella osteomyelitis in itself is predominantly seen in patients with haemoglobinopathies such as sickle cell disease or thalassemia...

  19. Development and evaluation of DNA and RNA real-time assays for food analysis using the hilA gene of Salmonella enterica subspecies enterica.

    Science.gov (United States)

    McCabe, Evonne M; Burgess, Catherine M; O'Regan, Edel; McGuinness, Sheila; Barry, Thomas; Fanning, Séamus; Duffy, Geraldine

    2011-05-01

    The objective of this study was the development of DNA and RNA real-time PCR methods for detection of food-borne Salmonella sp. as rapid alternatives to the traditional cultural method (ISO 6579, 2004) in fresh meat carcasses and processed meat samples. These PCR methods were based on the hilA sequence, with primers and hybridisation probes designed against this gene target. The primers and probes were evaluated for their efficiency and dynamic range and subsequently the specificity of the assay was tested using 106 Salmonella enterica subspecies enterica strains and 30 non-salmonellae strains. An internal amplification control (IAC) was also developed for incorporation. The optimum copy number of IAC was determined to be 500 copies per reaction. A complementary enrichment protocol was adapted from the existing standard ISO 6579:2004 and consisted of enrichment in Buffered Peptone Water (BPW) 22 ± 2 h and a second selective enrichment for 6 h in Rappaport Vassiliadis with Soya (RVS). The DNA and RNA-based real-time PCR protocols, were applied to meat samples inoculated with Salmonella enterica subspecies enterica strains, including swabs from meat carcasses and minced beef samples which were heat treated or frozen. The developed methods have the potential as useful alternatives to the standard ISO 6579:2004 method for the detection of Salmonella enterica subspecies enterica on carcass swabs and raw meat using hilA as a target. The DNA assay is a useful tool for the screening of meat samples in the abattoir within 3 days of slaughter or in a food production process and the RNA-based assay has the potential to detect viable Salmonella enterica subspecies enterica in ready-to-eat products. Copyright © 2010 Elsevier Ltd. All rights reserved.

  20. Genomic characterization of an extensively-drug resistance Salmonella enterica serotype Indiana strain harboring blaNDM-1 gene isolated from a chicken carcass in China.

    Science.gov (United States)

    Wang, Wei; Peng, Zixin; Baloch, Zulqarnain; Hu, Yujie; Xu, Jin; Zhang, Wenhui; Fanning, Séamus; Li, Fengqin

    2017-11-01

    The objective of this study was to genetically characterize the antimicrobial resistance mechanisms of Salmonella enterica serotype Indiana C629 isolated from a chicken carcass in China in 2014. Antimicrobial susceptibility against a panel of 23 antimicrobial agents was carried out on Salmonella enterica serotype Indiana C629 and assessed according to CLSI standards. Whole-genome sequencing of this isolate was conducted to obtain the complete genome of S. Indiana. Salmonella Indiana C629 expressed an XDR phenotype being resistant to more than 20 antimicrobial agents, including imipenem and meropenem. From the analysis of the resistance mechanisms, two mutations were identified in subunit A of DNA gyrase within the quinolone resistance determining region, in addition to the acquisition of mobile efflux pumps encoding oqxA/B/R. Additionally, four beta-lactamases resistance genes (blaCTX-M-65, blaTEM-1, blaOXA-1, and blaNDM-1), five aminoglycosides resistance genes (aac(3)-IV, aac(6')-Ib-cr, aadA2, aadA5, and aph(4)-Ia), two phenicol resistance genes (catB3 and floR), and five trimethoprim/sulfamethoxazole resistance genes (sul1/2/3 and dfrA12/17) were also identified. A total of 191 virulence genes were identified. Among them, 57 belonged to type-three secretion system (T3SS) encoding genes, 55 belonged to fimbrial adherence encoding genes, and 39 belonged to flagella-encoding genes CONCLUSIONS: This study demonstrated that multi-resistance mechanisms consistent with an XDR-phenotype, along with various virulence encoding genes of a S. Indiana strain in China These findings highlight the importance of cooperation among different sectors in order to monitor the spread of resistant pathogens among food animal, foods of animal origin and human beings that might further take measures to protect consumers' health. Copyright © 2017 Elsevier GmbH. All rights reserved.

  1. Epidemic multiple drug resistant Salmonella Typhimurium causing invasive disease in sub-Saharan Africa have a distinct genotype.

    Science.gov (United States)

    Kingsley, Robert A; Msefula, Chisomo L; Thomson, Nicholas R; Kariuki, Samuel; Holt, Kathryn E; Gordon, Melita A; Harris, David; Clarke, Louise; Whitehead, Sally; Sangal, Vartul; Marsh, Kevin; Achtman, Mark; Molyneux, Malcolm E; Cormican, Martin; Parkhill, Julian; MacLennan, Calman A; Heyderman, Robert S; Dougan, Gordon

    2009-12-01

    Whereas most nontyphoidal Salmonella (NTS) are associated with gastroenteritis, there has been a dramatic increase in reports of NTS-associated invasive disease in sub-Saharan Africa. Salmonella enterica serovar Typhimurium isolates are responsible for a significant proportion of the reported invasive NTS in this region. Multilocus sequence analysis of invasive S. Typhimurium from Malawi and Kenya identified a dominant type, designated ST313, which currently is rarely reported outside of Africa. Whole-genome sequencing of a multiple drug resistant (MDR) ST313 NTS isolate, D23580, identified a distinct prophage repertoire and a composite genetic element encoding MDR genes located on a virulence-associated plasmid. Further, there was evidence of genome degradation, including pseudogene formation and chromosomal deletions, when compared with other S. Typhimurium genome sequences. Some of this genome degradation involved genes previously implicated in virulence of S. Typhimurium or genes for which the orthologs in S. Typhi are either pseudogenes or are absent. Genome analysis of other epidemic ST313 isolates from Malawi and Kenya provided evidence for microevolution and clonal replacement in the field.

  2. Molecular detection of Rickettsia typhi in cats and fleas.

    Directory of Open Access Journals (Sweden)

    Maria Mercedes Nogueras

    Full Text Available BACKGROUND: Rickettsiatyphi is the etiological agent of murine typhus (MT, a disease transmitted by two cycles: rat-flea-rat, and peridomestic cycle. Murine typhus is often misdiagnosed and underreported. A correct diagnosis is important because MT can cause severe illness and death. Our previous seroprevalence results pointed to presence of human R. typhi infection in our region; however, no clinical case has been reported. Although cats have been related to MT, no naturally infected cat has been described. The aim of the study is to confirm the existence of R. typhi in our location analyzing its presence in cats and fleas. METHODOLOGY/PRINCIPAL FINDINGS: 221 cats and 80 fleas were collected from Veterinary clinics, shelters, and the street (2001-2009. Variables surveyed were: date of collection, age, sex, municipality, living place, outdoor activities, demographic area, healthy status, contact with animals, and ectoparasite infestation. IgG against R. typhi were evaluated by indirect immunofluorescence assay. Molecular detection in cats and fleas was performed by real-time PCR. Cultures were performed in those cats with positive molecular detection. Statistical analysis was carried out using SPSS. A p < 0.05 was considered significant. Thirty-five (15.8% cats were seropositive. There were no significant associations among seropositivity and any variables. R. typhi was detected in 5 blood and 2 cultures. High titres and molecular detection were observed in stray cats and pets, as well as in spring and winter. All fleas were Ctenocephalides felis. R. typhi was detected in 44 fleas (55%, from shelters and pets. Co-infection with R. felis was observed. CONCLUSIONS: Although no clinical case has been described in this area, the presence of R. typhi in cats and fleas is demonstrated. Moreover, a considerable percentage of those animals lived in households. To our knowledge, this is the first time R. typhi is detected in naturally infected cats.

  3. Feeding a high dosage of zinc oxide affects suppressor of cytokine gene expression in Salmonella Typhimurium infected piglets.

    Science.gov (United States)

    Schulte, Jasper N; Brockmann, Gudrun A; Kreuzer-Redmer, Susanne

    2016-10-01

    Suppressor of cytokine signaling (SOCS) proteins play an important role in the regulation of the immune response by inhibiting cytokines. Here we investigated the effects of zinc oxide fed at three different dosages (LZN=57ppm, MZN=167ppm, HZN=2425ppm) to weaned piglets that were or were not orally infected with Salmonella enterica serovar Typhimurium DT 104. We detected higher expression of SOCS3 six days after weaning for all analyzed piglets, regardless of the infection or the zinc feeding, suggesting a stress induced immune response. Whereas, SOCS1 showed only higher transcript amounts in S. Typhimurium infected piglets, especially the LZN group. This might indicate an infection regulating effect of zinc oxide in the infection model. After 42days of infection, the expression of SOCS2, SOCS4, and SOCS7 was increased only in animals fed the highest concentrations of zinc oxide, while non-infected piglets at the age of 56days showed no regulation for these genes. The up-regulation of SOCS genes in the mesenteric lymph nodes of piglets fed a diet with a very high concentration of zinc over 6 weeks suggests that such treatments may impair the immune response. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. 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

    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...

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

    DEFF Research Database (Denmark)

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

    1998-01-01

    to the sul1 and qacE Delta 1 genes characteristic of integrons. The first integron encoded the ant (3")-Ia gene that specified resistance to spectinomycin and streptomycin. The second contained the pse-1 beta-lactamase gene. All the multiresistant strains contained both integrons. The presence of these two...

  6. Antimicrobial susceptibility and occurrence of resistance genes among Salmonella enterica serovar Weltevreden from different countries

    DEFF Research Database (Denmark)

    Aarestrup, Frank Møller; Lertworapreecha, M.; Evans, M.C.

    2003-01-01

    and gentamicin. All nine ampicillin-resistant isolates contained a sequence similar to the bla(TEM-1b) gene, one of the eight chloramphenicol-resistant isolates a sequence similar to the catA1 gene, all three neomycin-resistant isolates a sequence similar to the aphA-2 gene, 16 (73%) of the 22 streptomycin...

  7. The consequences of a sudden demographic change on the seroprevalence pattern, virulence genes, identification and characterisation of integron-mediated antibiotic resistance in the Salmonella enterica isolated from clinically diarrhoeic humans in Egypt.

    Science.gov (United States)

    Osman, K M; Hassan, W M M; Mohamed, R A H

    2014-08-01

    The present study was undertaken to identify and characterise integrons and integrated resistance gene cassettes among eight multidrug-resistant (MDR) Salmonella serovars isolated from humans in Egypt. Virulotyping by polymerase chain reaction (PCR) was used for the detection of the presence of virulence genes. Integron PCR was used to detect the presence of class 1 in the MDR strains. The associated individual resistance gene cassettes were identified using specific PCRs. The isolated serovars were Salmonella Grampian (C1; 2/5), Larose (C1; 1/5), Hato (B; 1/5) and Texas (B; 1/5). Among the Salmonella serovars, five Salmonella isolates showed the highest resistance to amoxicillin, ampicillin, chloramphenicol, lincomycin, gentamicin, nalidixic acid, streptomycin and trimethoprim (100%), followed by neomycin, norfloxacin and tetracycline (80%), while the lowest resistance was recorded to colistin sulphate and ciprofloxacin in percentages of 20 and 40%, respectively. The invA, avrA, ssaQ, mgtC, siiD and sopB genes were detected in all isolates (100%), while the spvC and gipA genes were totally (100%) absent from all isolates. The remaining three virulence genes were diversely distributed as follows: the bcfC gene was detected in all isolates except Salmonella Hato (80%); the sodC1 gene was detected only in Salmonella Grampian and Salmonella Texas (60%); and the sopE1 gene was detected only in Salmonella Grampian, Hato and Texas (60%). Class 1 integrons were detected in 90% of the MDR isolates, comprising serovars Muenster, Florian, Noya, Grampian, Larose, Hato and Texas. Of the class 1 integron-positive isolates, 45% harboured Salmonella genomic island 1 (SGI1) either right junction or right and left junction having an A-C-S-T phenotype. Of the class 1 integron-positive isolates, 44% harboured integron gene cassette aadA2, while 11% harboured the floR gene present in multidrug resistance flanked by two integrons of SGI1. The results of the present study indicate that

  8. First Description of the Extended Spectrum-Beta-Lactamase Gene blaCTX-M-109 in Salmonella Grumpensis Strains Isolated from Neonatal Nosocomial Infections in Dakar, Senegal.

    Directory of Open Access Journals (Sweden)

    Amadou Diop

    Full Text Available Nosocomial infections are very common in African hospitals, particularly in neonatal units. These infections are most often caused by bacteria such as Escherichia coli, Klebsiella spp and Staphylococcus spp. Salmonella strains are rarely involved in nosocomial infections. Here, we report the first description of S. Grumpensis in neonatal infections in Senegal. Seventeen Salmonella strains were isolated from hospitalized infants' stool samples. The following resistance phenotype was described in strains: AMXRTICRCFR FOXRCFXRCTXRCAZRIMPSATMRNARNORRCIPRTMRGMRTERSXTR. All isolates were susceptible to imipenem, 15 out of 17 produced an extended spectrum ß-lactamase (ESBL. blaOXA-1, blaSHV-1, blaTEM-1, blaCTX-M1 genes were detected in strains 8, 13, 5 and 8, respectively. blaCTX-M1 sequencing revealed the presence of blaCTX-M-109. Thirteen of the 17 Salmonella Grumpensis strains were analyzed by PFGE. These 13 isolates belonged to a single pulsotype and were genotypically identical. This is the first report of neonatal S. Grumpensis infections in Senegal, and the first report of blaCTX-M-109 in the genus Salmonella.

  9. Plasmid- and chromosomal genes-encoded two separate O-polysaccharide chains of Salmonella Uccle (O:3,54)--structural elucidation.

    Science.gov (United States)

    Paszkiewicz, Monika; Tokarska-Pietrzak, Ewa; Gołębiowski, Marek; Kunikowska, Danuta; Stepnowski, Piotr

    2013-11-01

    The bacterium Salmonella Uccle belongs to serotype O:54 in the Kauffmann'a-White scheme. Group O:54 is unique among the serogroups belonging to the genus Salmonella. Normally, the enzymes involved in the biosynthesis of the repeating units of somatic antigen are encoded by a set of genes, located in the region of the bacterial chromosome. Expression of O54 factor is associated with the presence of the plasmid. Factor O54 can be lost spontaneously in the subcultures of some serotypes. In these cases, the O54 negative variants become indistinguishable from the serotypes classified in other groups. We noticed lower activity of LPS-u O:3,54 with rabbit sera against antigens O:3, when compared with the activity of sera anti-A:54, which may indicate a partial inhibition of the expression of factor O:3 on the surface of the bacterial cell. The main aim of our study was to answer the question whether the products of different biosynthetic pathways combine on the outer side of the cytoplasmic membrane, thus forming a single chain or separate chains. Therefore, the O-polysaccharides (O-antigen) of Salmonella Uccle O:3,54 were isolated by mild acid degradation of both obtained LPSs and their structure was established using sugar and methylation analysis and NMR spectroscopy. The primary structure of two separate O-polysaccharide chains isolated from Salmonella Uccle were established. Copyright © 2013. Published by Elsevier Inc.

  10. Lymphocytic proliferative response to outer-membrane proteins isolated from Salmonella.

    Science.gov (United States)

    González, C R; Isibasi, A; Ortiz-Navarrete, V; Paniagua, J; García, J A; Blanco, F; Kumate, J

    1993-01-01

    Porins isolated from Salmonella typhi have been demonstrated to protect against the challenge with this bacteria in mice. The mechanism has not been clarified, but could be associated with activation of both humoral and cellular immunity. In order to evaluate the induction of specific T cell responses, the lymphocytic proliferation to porins isolated from Salmonella typhimurium, Salmonella typhi and Escherichia coli was examined by 3H-thymidine incorporation assay in mice immunized with three different antigens: acetone-killed S. typhimurium, its porins, or outer-membrane proteins (OMPs) isolated from S. typhi. Higher proliferative responses were observed in mice immunized with porins and OMPs compared with those which received the acetone-killed bacteria. Although cross-reactivity was observed between porins, they were not mitogenic. Moreover, porins were able to activate T lymphocytes isolated from mice immunized with S. typhi OMPs. These results suggest that T cell activation, through the release of lymphokines, may play a role in the induction of protective immunity with porins.

  11. 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

    Directory of Open Access Journals (Sweden)

    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

  12. Salmonella Infections (For Parents)

    Science.gov (United States)

    ... Needs a Kidney Transplant Vision Facts and Myths Salmonella Infections KidsHealth > For Parents > Salmonella Infections Print A ... Last? Can Salmonella Infections Be Prevented? What Is Salmonella ? Salmonella is a kind of bacteria , with many ...

  13. Horizontal gene transfer and antibiotic resistance plasmids in multi-drug resistant Salmonella enterica serovars

    Science.gov (United States)

    Antibiotic resistant foodborne pathogens pose serious public health concerns and increase the burden of disease treatment. Antibiotic resistance genes can reside on the bacterial chromosome or on other self-replicating DNA molecules such as plasmids. The resistance genes/DNA can be transferred int...

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

    DEFF Research Database (Denmark)

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

    2014-01-01

    -td was shown not to affect invasion of cultured epithelial cells, while the absence of the gene significantly affects uptake and intracellular survival within macrophages. The gene st313-td was proven to be strongly associated to invasiveness, harboured by 92.5% of S. Typhimurium blood isolates (n...

  15. Rickettsia typhi and R. felis in rat fleas (Xenopsylla cheopis), Oahu, Hawaii.

    Science.gov (United States)

    Eremeeva, Marina E; Warashina, Wesley R; Sturgeon, Michele M; Buchholz, Arlene E; Olmsted, Gregory K; Park, Sarah Y; Effler, Paul V; Karpathy, Sandor E

    2008-10-01

    Rickettsia typhi (prevalence 1.9%) and R. felis (prevalence 24.8%) DNA were detected in rat fleas (Xenopsylla cheopis) collected from mice on Oahu Island, Hawaii. The low prevalence of R. typhi on Oahu suggests that R. felis may be a more common cause of rickettsiosis than R. typhi in Hawaii.

  16. Rickettsia typhi and R. felis in Rat Fleas (Xenopsylla cheopis), Oahu, Hawaii

    OpenAIRE

    Eremeeva, Marina E.; Warashina, Wesley R.; Sturgeon, Michele M.; Buchholz, Arlene E.; Olmsted, Gregory K.; Park, Sarah Y.; Effler, Paul V.; Karpathy, Sandor E.

    2008-01-01

    Rickettsia typhi (prevalence 1.9%) and R. felis (prevalence 24.8%) DNA were detected in rat fleas (Xenopsylla cheopis) collected from mice on Oahu Island, Hawaii. The low prevalence of R. typhi on Oahu suggests that R. felis may be a more common cause of rickettsiosis than R. typhi in Hawaii.

  17. Enhancement of Gastric Ulcer Healing and Angiogenesis by Hepatocyte Growth Factor Gene Mediated by Attenuated Salmonella in Rats.

    Science.gov (United States)

    Ha, Xiaoqin; Peng, Junhua; Zhao, Hongbin; Deng, Zhiyun; Dong, Juzi; Fan, Hongyan; Zhao, Yong; Li, Bing; Feng, Qiangsheng; Yang, Zhihua

    2017-02-01

    The present study developed an oral hepatocyte growth factor (HGF) gene therapy strategy for gastric ulcers treatment. An attenuated Salmonella typhimurium that stably expressed high HGF (named as TPH) was constructed, and the antiulcerogenic effect of TPH was evaluated in a rat model of gastric ulcers that created by acetic acid subserosal injection. From day 5 after injection, TPH (1 × 10⁹ cfu), vehicle (TP, 1 × 10⁹ cfu), or sodium bicarbonate (model control) was administered orally every alternate day for three times. Then ulcer size was measured at day 21 after ulcer induction. The ulcer area in TPH-treated group was 10.56 ± 3.30 mm², which was smaller when compared with those in the TP-treated and model control groups (43.47 ± 4.18 and 56.25 ± 6.38 mm², respectively). A higher level of reepithelialization was found in TPH-treated group and the crawling length of gastric epithelial cells was significantly longer than in the other two groups (P ulcer granulation tissues of the TPH-treated rats was 39.9 vessels/mm², which was greater than in the TP-treated and model control rats, with a significant statistical difference. These results suggest that TPH treatment significantly accelerates the healing of gastric ulcers via stimulating proliferation of gastric epithelial cells and enhancing angiogenesis on gastric ulcer site.

  18. Altered virulence potential of Salmonella Enteritidis cultured in different foods: A cumulative effect of differential gene expression and immunomodulation.

    Science.gov (United States)

    Jaiswal, Sangeeta; Sahoo, Prakash Kumar; Ryan, Daniel; Das, Jugal Kishore; Chakraborty, Eesha; Mohakud, Nirmal Kumar; Suar, Mrutyunjay

    2016-08-02

    Salmonella enterica serovars Enteritidis (S. Enteritidis) is one of the most common causes of food borne illness. Bacterial growth environment plays an important role in regulating gene expression thereby affecting the virulence profile of the bacteria. Different foods present diverse growth conditions which may affect the pathogenic potential of the bacteria. In the present study, the effect of food environments on the pathogenic potential of S. Enteritidis has been evaluated. S. Enteritidis was grown in different foods e.g. egg white, peanut butter and milk, and virulent phenotypes were compared to those grown in Luria Bertani broth. In-vivo experiments in C57BL/6 mice revealed S. Enteritidis grown in egg white did not induce significant (panalysis revealed SPI-1 effectors were downregulated in bacteria grown in egg white. Interestingly, bacteria grown in egg white showed reversal of phenotype upon change in growth media to LB. Additionally, bacteria grown in milk and peanut butter showed different degrees of virulence in mice as compared to those grown in LB media. Thus, the present study demonstrates that, S. Enteritidis grown in egg white colonizes systemic sites without causing colitis in a mouse model, while bacteria grown in milk and peanut butter show different pathogenicity profiles suggesting that food environments significantly affect the pathogenicity of S. Enteritidis. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Salmonella: Salmonellosis

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Hansen, Trine; Maurischat, Sven

    2015-01-01

    Salmonella remains one of the most important zoonotic pathogenic bacteria and is the causative agents of salmonellosis. The aim of this article is to give an overview of Salmonella and salmonellosis, starting by describing the characteristics of the microorganism Salmonella, including biochemical...... properties, physiology, classification, and nomenclature. Thereafter, the epidemiology of the organism is introduced, including the routes of transmission. Finally, the disease salmonellosis, the virulence mechanisms, and the occurrence in different types of food are described....

  20. Characterization of integrons and resistance genes in multidrug-resistant Salmonella enterica isolated from meat and dairy products in Egypt.

    Science.gov (United States)

    Ahmed, Ashraf M; Shimamoto, Toshi; Shimamoto, Tadashi

    2014-10-17

    Foodborne pathogens are a leading cause of illness and death, especially in developing countries. The problem is exacerbated if bacteria attain multidrug resistance. Little is currently known about the extent of antibiotic resistance in foodborne pathogens and the molecular mechanisms underlying this resistance in Africa. Therefore, the current study was carried out to characterize, at the molecular level, the mechanism of multidrug resistance in Salmonella enterica isolated from 1600 food samples (800 meat products and 800 dairy products) collected from different street venders, butchers, retail markets and slaughterhouses in Egypt. Forty-seven out of 69 isolates (68.1%) showed multidrug resistance phenotypes to at least three classes of antimicrobials. The incidence of multidrug-resistant isolates was higher in meat products (37, 69.8%) than in dairy products (10, 62.5%). The multidrug-resistant serovars included, S. enterica serovar Typhimurium (24 isolates, 34.8%), S. enterica serovar Enteritidis, (15 isolates, 21.8%), S. enterica serovar Infantis (7 isolates, 10.1%) and S. enterica non-typable serovar (1 isolate, 1.4%). The highest resistance was to ampicillin (95.7%), then to kanamycin (93.6%), spectinomycin (93.6%), streptomycin (91.5%) and sulfamethoxazole/trimethoprim (91.5%). PCR and DNA sequencing were used to screen and characterize integrons and antibiotic resistance genes and 39.1% and 8.7% of isolates were positive for class 1 and class 2 integrons, respectively. β-lactamase-encoding genes were identified in 75.4% of isolates and plasmid-mediated quinolone resistance genes were identified in 27.5% of isolates. Finally, the florphenicol resistance gene, floR, was identified in 18.8% of isolates. PCR screening identified S. enterica serovar Typhimurium DT104 in both meat and dairy products. This is the first study to report many of these resistance genes in dairy products. This study highlights the high incidence of multidrug-resistant S. enterica in

  1. Plasmid-mediated quinolone resistance by genes qnrA1 and qnrB19 in Salmonella strains isolated in Brazil.

    Science.gov (United States)

    Ferrari, Rafaela; Galiana, Antonio; Cremades, Rosa; Rodriguez, Juan Carlos; Magnani, Marciane; Tognim, Maria Cristina B; Oliveira, Tereza C R M; Royo, Gloria

    2011-07-04

    Considering the importance of the mechanisms involved in quinolone resistance, this study evaluate the presence of PMQR in 126 epidemic and not epidemic strains of Salmonella spp. It was noted that presence of PMQR, by itself, did not generate resistance to ciprofloxacin; but detection of qnr genes in Salmonella spp. and the identification of the qnrB19 variant in a strain of poultry origin alert for the indiscriminate use of quinolones in poultry production, that can result in a pressure for mutant selection of resistant strains with a clinical limitation use of FQs in the near future. And last but not least, is the need to continued study of resistance mechanisms and to monitor the microbial resistance profile of epidemiological strains.

  2. Scarless deletion of up to seven methyl-accepting chemotaxis genes with an optimized method highlights key function of CheM in Salmonella Typhimurium.

    Directory of Open Access Journals (Sweden)

    Stefanie Hoffmann

    Full Text Available Site-directed scarless mutagenesis is an essential tool of modern pathogenesis research. We describe an optimized two-step protocol for genome editing in Salmonella enterica serovar Typhimurium to enable multiple sequential mutagenesis steps in a single strain. The system is based on the λ Red recombinase-catalyzed integration of a selectable antibiotics resistance marker followed by replacement of this cassette. Markerless mutants are selected by expressing the meganuclease I-SceI which induces double-strand breaks in bacteria still harboring the resistance locus. Our new dual-functional plasmid pWRG730 allows for heat-inducible expression of the λ Red recombinase and tet-inducible production of I-SceI. Methyl-accepting chemotaxis proteins (MCP are transmembrane chemoreceptors for a vast set of environmental signals including amino acids, sugars, ions and oxygen. Based on the sensory input of MCPs, chemotaxis is a key component for Salmonella virulence. To determine the contribution of individual MCPs we sequentially deleted seven MCP genes. The individual mutations were validated by PCR and genetic integrity of the final seven MCP mutant WRG279 was confirmed by whole genome sequencing. The successive MCP mutants were functionally tested in a HeLa cell infection model which revealed increased invasion rates for non-chemotactic mutants and strains lacking the MCP CheM (Tar. The phenotype of WRG279 was reversed with plasmid-based expression of CheM. The complemented WRG279 mutant showed also partially restored chemotaxis in swarming assays on semi-solid agar. Our optimized scarless deletion protocol enables efficient and precise manipulation of the Salmonella genome. As demonstrated with whole genome sequencing, multiple subsequent mutagenesis steps can be realized without the introduction of unwanted mutations. The sequential deletion of seven MCP genes revealed a significant role of CheM for the interaction of S. Typhimurium with host cells

  3. Multiple transmissible genes encoding fluoroquinolone and third-generation cephalosporin resistance co-located in non-typhoidal Salmonella isolated from food-producing animals in China.

    Science.gov (United States)

    Jiang, Hong-Xia; Song, Li; Liu, Ji; Zhang, Xiao-Hua; Ren, Yan-Na; Zhang, Wen-Hui; Zhang, Jing-Yuan; Liu, Ya-Hong; Webber, Mark A; Ogbolu, David O; Zeng, Zhen-Ling; Piddock, Laura J V

    2014-03-01

    The aim of this study was to identify genes conferring resistance to fluoroquinolones and extended-spectrum β-lactams in non-typhoidal Salmonella (NTS) from food-producing animals in China. In total, 31 non-duplicate NTS were obtained from food-producing animals that were sick. Isolates were identified and serotyped and the genetic relatedness of the isolates was determined by pulsed-field gel electrophoresis of XbaI-digested chromosomal DNA. Antimicrobial susceptibility was determined using Clinical and Laboratory Standards Institute methodology. The presence of extended-spectrum β-lactamase (ESBL) and fluoroquinolone resistance genes was established by PCR and sequencing. Genes encoded on transmissible elements were identified by conjugation and transformation. Plasmids were typed by PCR-based replicon typing. The occurrence and diversity of numerous different transmissible genes conferring fluoroquinolone resistance [qnrA, qnrD, oqxA and aac(6')-Ib-cr] and ESBLs (CTX-M-27 and CTX-M-14), and which co-resided in different isolates and serovars of Salmonella, were much higher than in European countries. Furthermore, different plasmids encoded fluoroquinolone resistance (ca. 6 kb) and β-lactam resistance (ca. 63 kb) and these co-resided in isolates with mutations in topoisomerase genes (gyrA and parC) giving very resistant Salmonella. The presence of multidrug-resistant bacteria in food-producing animals in countries that export foodstuffs suggests that global transfer of antibiotic resistances from country to country on food is possible. Copyright © 2013 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  4. Influence of Salmonella enterica serovar Pullorum pathogenicity island 2 on type III secretion system effector gene expression in chicken macrophage HD11 cells.

    Science.gov (United States)

    Yin, Junlei; Chen, Yun; Xie, Xiaolei; Xia, Jie; Li, Qiuchun; Geng, Shizhong; Jiao, Xinan

    2017-04-01

    Salmonella pathogenicity island 2 (SPI2) can encode type III secretion system 2 (T3SS2) which plays an important role in systemic disease development through delivering different effector proteins into host cells. Here, the influence of Salmonella Pullorum pathogenicity island 2 on T3SS2 effector gene expression was studied using qRT-PCR in chicken macrophage HD11 cells. Our results showed that all the detected genes (including pseudogenes sifB, sspH2 and steC) can express in HD11 cells of S. Pullorum infection; deletion of SPI2 of S. Pullorum did not significantly affect the expression of genes cigR, gtgA, slrP, sopD, sseK1, steB and steC, but had a significant effect on the expression of genes pipB2, sifB, sopD2, sseJ, sseL, sspH2, steD, sifA, pipB and steA at different degrees. These results suggest that SPI2 can significantly affect the expression of some T3SS2 effector genes. Some effectors may have secretion pathways other than T3SS2 and pseudogenes may play roles in the process of S. Pullorum infection.

  5. Poor biofilm-forming ability and long-term survival of invasive Salmonella Typhimurium ST313.

    Science.gov (United States)

    Ramachandran, Girish; Aheto, Komi; Shirtliff, Mark E; Tennant, Sharon M

    2016-07-01

    Salmonella enterica serovar Typhimurium, an enteric pathogen that causes a self-limiting gastroenteritis, forms biofilms on different surfaces. In sub-Saharan Africa, Salmonella Typhimurium of a novel sequence type (ST) 313 was identified and produces septicemia in the absence of gastroenteritis. No animal reservoir has been identified, and it is hypothesized that transmission occurs via human to human. In this study, we show that invasive Salmonella Typhimurium ST313 strains from Mali are poor biofilm producers compared to Salmonella Typhimurium ST19 strains, which are found worldwide and are known to be associated with gastroenteritis. We evaluated biofilms using crystal violet staining, examination of the red, dry and rough morphotype, pellicle formation and a continuous flow system. One month-old Salmonella Typhimurium ST19 colonies survived in the absence of exogenous nutrients and were highly resistant to sodium hypochlorite treatment compared to Salmonella Typhimurium ST313. This study for the first time demonstrates the comparative biofilm-forming ability and long-term survival of clinical Salmonella Typhimurium ST19 and ST313 isolates. Salmonella Typhimurium ST19 strains are strong biofilm producers and can survive desiccation compared to Salmonella Typhimurium ST313 that form weak biofilms and survive poorly following desiccation. Our data suggest that like Salmonella Typhi, Salmonella Typhimurium ST313 lack mechanisms that allow it to persist in the environment. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Selected Lactic Acid-Producing Bacterial Isolates with the Capacity to Reduce Salmonella Translocation and Virulence Gene Expression in Chickens

    OpenAIRE

    Xiaojian Yang; Jennifer Brisbin; Hai Yu; Qi Wang; Fugui Yin; Yonggang Zhang; Parviz Sabour; Shayan Sharif; Joshua Gong

    2014-01-01

    BACKGROUND: Probiotics have been used to control Salmonella colonization/infection in chickens. Yet the mechanisms of probiotic effects are not fully understood. This study has characterized our previously-selected lactic acid-producing bacterial (LAB) isolates for controlling Salmonella infection in chickens, particularly the mechanism underlying the control. METHODOLOGY/PRINCIPAL FINDINGS: In vitro studies were conducted to characterize 14 LAB isolates for their tolerance to low pH (2.0) an...

  7. Invasive Salmonella infections in areas of high and low malaria transmission intensity in Tanzania.

    Science.gov (United States)

    Biggs, Holly M; Lester, Rebecca; Nadjm, Behzad; Mtove, George; Todd, Jim E; Kinabo, Grace D; Philemon, Rune; Amos, Ben; Morrissey, Anne B; Reyburn, Hugh; Crump, John A

    2014-03-01

     The epidemiology of Salmonella Typhi and invasive nontyphoidal Salmonella (NTS) differs, and prevalence of these pathogens among children in sub-Saharan Africa may vary in relation to malaria transmission intensity.  We compared the prevalence of bacteremia among febrile pediatric inpatients aged 2 months to 13 years recruited at sites of high and low malaria endemicity in Tanzania. Enrollment at Teule Hospital, the high malaria transmission site, was from June 2006 through May 2007, and at Kilimanjaro Christian Medical Centre (KCMC), the low malaria transmission site, from September 2007 through August 2008. Automated blood culture, malaria microscopy with Giemsa-stained blood films, and human immunodeficiency virus testing were performed.  At Teule, 3639 children were enrolled compared to 467 at KCMC. Smear-positive malaria was detected in 2195 of 3639 (60.3%) children at Teule and 11 of 460 (2.4%) at KCMC (P Salmonella Typhi was isolated from 11 (0.3%) children at Teule and 6 (1.3%) at KCMC (P = .008). With NTS excluded, the prevalence of bacteremia at Teule was 5.0% and at KCMC 4.1% (P = .391).  Where malaria transmission was intense, invasive NTS was common and Salmonella Typhi was uncommon, whereas the inverse was observed at a low malaria transmission site. The relationship between these pathogens, the environment, and the host is a compelling area for further research.

  8. SALMONELLA SPECIES

    African Journals Online (AJOL)

    DR. AMINU

    e. Biochemical screening and serological tests for Salmonellae. Identification of Salmonella species was done biochemically. Triple sugar Iron (TSI) agar motility, urease and citrate utilization tests were also used to screen the isolates before serologic testing was performed. (Cheesbrough, 2002; Perilla, 2003). Triple sugar ...

  9. Direct evidence of Rickettsia typhi infection in Rhipicephalus ...

    African Journals Online (AJOL)

    Murine typhus is a rickettsiosis caused by Rickettsia typhi, whose transmission is carried out by rat fleas in urban settlements as classically known, but it also has been related to cat fleas in a sub-urban alternative cycle that has been suggested by recent reports. These studies remarks that in addition to rats, other animals ...

  10. Identification of a novel transposon-associated phosphoethanolamine transferase gene, mcr-5, conferring colistin resistance in d-tartrate fermenting Salmonella enterica subsp. enterica serovar Paratyphi B

    DEFF Research Database (Denmark)

    Borowiak, Maria; Fischer, Jennie; Hammerl, Jens A

    2017-01-01

    phosphoethanolamine transferase genes involved in colistin resistance. Subsequently PCR screening, S1-PFGE and DNA-DNA hybridization were performed to analyse the prevalence and location of the identified mcr-5 gene. Cloning and transformation experiments in Escherichia coli DH5α and Salmonella Paratyphi B d Ta...... during 2011-13. The respective gene, further termed as mcr-5 (1644 bp), is part of a 7337 bp transposon of the Tn 3 family and usually located on related multi-copy ColE-type plasmids. Interestingly, in one isolate an additional subclone with a chromosomal location of the mcr-5 transposon was observed....... Our findings suggest that the transfer of colistin-resistance-mediating phosphoethanolamine transferase genes from bacterial chromosomes to mobile genetic elements has occurred in multiple independent events raising concern regarding their variety, prevalence and impact on public health....

  11. Study of antibiotic resistance pattern and incidence of pathogenic genes of mgtC, spi4R, agfA, invE/A and ttrC in Salmonella infantis isolated from clinical specimens

    Directory of Open Access Journals (Sweden)

    Roya Aghdasi-Araghinezhad

    2017-12-01

    Full Text Available Background: The importance of the health of red meat, poultry and eggs in human nutrition is very high. One of the factors that jeopardize the health of poultry food products is the bacterial family of Enterobacteriaceae, especially Salmonella. The aim of this study was to detect pathogenic genes in Salmonella infectious bacteria isolated from stool specimens using the multiple PCR assay. Materials and Methods: Selective and specific media for isolation of Salmonella were used. Primary isolation was carried out using Peptone water, Rapaport, selenite cysteine, MacConky agar and xylose-lysine deoxycholate agar. To confirm the diagnosis, biochemical tests including TSI, urea, endodontic, and citrate were used. The Salmonella Polyvalent Kit was used to determine Salmonella groups and mgtC, spi4R, agfA, invE/A and ttrC genes were studied in 60 samples by the multiple PCR method. Results: The results showed that all samples had 2 genes mgtC and ttrC, and none of the samples showed resistance to cefepime. Of the 60 samples of Salmonella, none were resistant to cefepime and ceftriaxone; 38.8% of the samples were resistant to amoxicillin, 53% to erythromycin and 38.3% to sulfamethoxazole. Conclusion: It can be concluded that cefepime is the best selective drug for the treatment of Salmonella infections. Identification and validation of genes in the region's bacteria can play a role in the broad epidemiological examination, antibiotic resistance, vaccine production, level of virulence, prevention and treatment. Also, evaluation of these genes in the samples for their virulence index is very important.

  12. Suppression of dendritic cell and T-cell activation by the pRST⁹⁸ Salmonella plasmid.

    Science.gov (United States)

    Wei, Li; Jin, Qili; Chu, Yuanyuan; Wu, Shuyan; Huang, Rui

    2015-03-01

    Salmonella evades host immune response via the expression of a variety of pathogenic factors. The 'pRST98' plasmid of Salmonella enterica serotype Typhi (S. Typhi) is involved in conferring the multidrug‑resistance and virulence of S. typhi. However, its specific effect on host‑cell function has remained elusive. Dendritic cells (DCs) are key regulators of immune responses. The present study therefore aimed to investigate whether pRST98 may target DCs involved in mediating the adaptive immune response. In vivo experiments with Salmonella enterica serotype Typhimurium χ3337 and χ3337/pRST98 revealed that pRST98 may influence multiple important functions of murine DCs, including maturation, survival and cytokine production. In addition, pRST98 markedly contributed to decreasing T‑cell activation. These data suggested that by targeting the aforementioned functions of DCs, pRST98 may partially overturn the adaptive immune defense mechanisms of the host, which are required for elimination of this pathogen from infected tissues. This may contribute to the evasion of host adaptive immune responses by S. Typhi and therefore provide a target for the prevention and treatment of typhoid fever.

  13. Antibodies in Action: Role of Human Opsonins in Killing Salmonella enterica Serovar Typhi▿

    Science.gov (United States)

    Lindow, Janet C.; Fimlaid, Kelly A.; Bunn, Janice Y.; Kirkpatrick, Beth D.

    2011-01-01

    Although vaccines have been available for over a century, a correlate of protection for typhoid fever has yet to be identified. Antibodies are produced in response to typhoid infection and vaccination and are generally used as the gold standard for determining vaccine immunogenicity, even though their role in clearance of Salmonella enterica serovar Typhi infections is poorly defined. Here, we describe the first functional characterization of S. Typhi-specific antibodies following vaccination with a new vaccine, M01ZH09 (Ty2 ΔaroC ΔssaV). We determined that postvaccination sera increased the uptake of wild-type S. Typhi by human macrophages up to 2.3-fold relative to prevaccination (day 0) or placebo samples. These results were recapitulated using immunoglobulins purified from postvaccination serum, demonstrating that antibodies were largely responsible for increases in uptake. Imaging verified that macrophages internalized 2- to 9.5-fold more S. Typhi when the bacteria were opsonized with postvaccination sera than when the bacteria were opsonized with day 0 or placebo sera. Once inside macrophages, the survival of S. Typhi was reduced as much as 50% when opsonized with postvaccination sera relative to day 0 or placebo serum samples. Lastly, bactericidal assays indicated that antibodies generated postvaccination were recognized by complement factors and assisted in killing S. Typhi: mean postvaccination bactericidal antibody titers were higher at all time points than placebo and day 0 titers. These data clearly demonstrate that there are at least two mechanisms by which antibodies facilitate killing of S. Typhi. Future work could lead to improved immunogenicity tests associated with vaccine efficacy and the identification of correlates of protection against typhoid fever. PMID:21628517

  14. Control of Invasive Salmonella Disease in Africa: Is There a Role for Human Challenge Models?

    Science.gov (United States)

    Gibani, Malick M; Jin, Celina; Darton, Thomas C; Pollard, Andrew J

    2015-11-01

    Invasive Salmonella disease in Africa is a major public health concern. With evidence of the transcontinental spread of the Salmonella Typhi H58 haplotype, improved estimates of the burden of infection and understanding of the complex interplay of factors affecting disease transmission are needed to assist with efforts aimed at disease control. In addition to Salmonella Typhi, invasive nontyphoidal Salmonella are increasingly recognized as an important cause of febrile illness and mortality in sub-Saharan Africa. Human experimental oral challenge studies with Salmonella can be used as a model to offer unique insights into host-pathogen interactions as well as a platform to efficiently test new diagnostic and vaccine candidates. In this article, we review the background and use of human challenge studies to date and discuss how findings from these studies may lead to progress in the control of invasive Salmonella disease in Africa. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.

  15. The microbiological and clinical characteristics of invasive salmonella in gallbladders from cholecystectomy patients in kathmandu, Nepal.

    Directory of Open Access Journals (Sweden)

    Sabina Dongol

    Full Text Available Gallbladder carriage of invasive Salmonella is considered fundamental in sustaining typhoid fever transmission. Bile and tissue was obtained from 1,377 individuals undergoing cholecystectomy in Kathmandu to investigate the prevalence, characteristics and relevance of invasive Salmonella in the gallbladder in an endemic area. Twenty percent of bile samples contained a Gram-negative organism, with Salmonella Typhi and Salmonella Paratyphi A isolated from 24 and 22 individuals, respectively. Gallbladders that contained Salmonella were more likely to show evidence of acute inflammation with extensive neutrophil infiltrate than those without Salmonella, corresponding with higher neutrophil and lower lymphocyte counts in the blood of Salmonella positive individuals. Antimicrobial resistance in the invasive Salmonella isolates was limited, indicating that gallbladder colonization is unlikely to be driven by antimicrobial resistance. The overall role of invasive Salmonella carriage in the gallbladder is not understood; here we show that 3.5% of individuals undergoing cholecystectomy in this setting have a high concentration of antimicrobial sensitive, invasive Salmonella in their bile. We predict that such individuals will become increasingly important if current transmission mechanisms are disturbed; prospectively identifying these individuals is, therefore, paramount for rapid local and regional elimination.

  16. Salmonella species on meat contact surfaces and processing water in Sokoto main market and abattoir, Nigeria

    Directory of Open Access Journals (Sweden)

    Olufemi Oludayo Faleke

    2017-03-01

    Full Text Available This study was carried out to determine Salmonella contamination of food contact surfaces and processing water in meat, fish and poultry processing units in Sokoto State, Nigeria. A total of 200 swab (100 from abattoir and 100 from poultry and fish markets and 60 processing water samples (30 from abattoir and 30 from poultry and fish markets were collected between May to August 2015. Cultural isolation, bio-typing and sero-grouping using Salmonella Sero-Quick Group Kit was conducted to analyse the samples. Seventy-five (75/260, 28.8 % of the total samples were positive to Salmonella by cultural isolation and bio-typing. Thirty (30/130; 23.1 % of samples collected in abattoir and 45 (45/130; 34.6 % of those collected from poultry and fish markets were positive for Salmonella respectively. Sero-groups D+Vi (Salmonella Typhi, B (Salmonella Paratyphi B, Salmonella Typhimurium and C (Salmonella Paratyphi C, Salmonella Cholerae suis were identified as the prevailing sero-groups in this study. Sero-group D+Vi has the highest prevalence (73.3 %; 55/75 from the positive bio-typing isolates. This study revealed the presence of contaminating and pathogenic Salmonella on food contact surfaces and processing water in the meat retail markets, indicating there is an urgent need to improve on the hygienic status of retail meat, poultry and fish markets.

  17. SEROTYPING AND ANTIMICROBIAL DRUG RESISTANCE OF SALMONELLA ISOLATED FROM LETTUCE AND HUMAN DIARRHEA SAMPLES IN BURKINA FASO.

    Science.gov (United States)

    Siourimè, Somda Namwin; Isidore, Bonkoungou Ouindgueta Juste; Oumar, Traoré; Nestor, Bassolé Ismael Henri; Yves, Traoré; Nicolas, Barro; Aly, Savadogo

    2017-01-01

    In Burkina Faso dirty water in particular those of the stoppings and the gutter ones are used for vegetables irrigation in the gardens. The aim of this study was to determine the prevalence and antibiotic susceptibility of Salmonella serotypes from humans and lettuce samples inBurkina Faso. Materials and Methods:Salmonella strains isolated from patients in 2009 to 2015 and lettuce samples in 2014 in Burkina Faso were serotyped using specific antisera. All strains were subjected to a set of 14 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method. Out of 154 Salmonella isolated, 60 were from human and 94 from lettuce samples. Serotyping revealed four different serotypes and 39% (60) untypeable strains from human and lettuce (14 and 46 strains). Salmonella serotypes from human and lettuce samples were: Paratyphi A (10% and 22%), Paratyphi B (34% and 8%), Paratyphi C (14% and 18%) and Typhi (21% and 1%). A high resistance of Salmonella Paratyphi B and Salmonella spp to tetracycline were 70% from human and 35 % from lettuce samples. Multiresistance was observed to tetracycline, chloramphenicol and amoxicillin/clavulanic-acid or ampicillin with Salmonella ParatyphiB 35% and Salmonella Typhi 33% from human samples and Salmonella spp 4% from lettuce samples. This study showed the diversity of Salmonella serotypes from both clinical and environmental samples and emergence of multiresistant Salmonella to antibiotics in Burkina Faso. A lettuce is a potential source of transmission of Salmonella causing diarrhea among human in Burkina Faso. List of non-standard Abbreviations : HDB: Hôpital du District de Bogodogo, LNSP: Laboratoire National de Santé Publique, DSG : District Sanitaire de Gourcy, DSB : District Sanitaire de Boromo.

  18. Distribution of virulence genes sefC, pefA and spvC in Salmonella Enteritidis phage type 4 strains isolated in Brazil Distribuição de genes de virulência sefC, pefA e spvC em cepas de Salmonella Enteritidis fago tipo 4 isoladas no Brasil

    Directory of Open Access Journals (Sweden)

    Karina Salvagni Castilla

    2006-06-01

    Full Text Available The distribution of virulence genes, sefC, pefA and spvC, was investigated in 110 Salmonella Enteritidis phage type 4 strains by polymerase chain reaction. Their influence in the caecal colonization and invasion of liver and spleen of one-day-old chickens was studied. Eight isolates were negative for the spvC gene, three for the pefA gene and one, for the sefC gene. These results allowed grouping the strains into four genotypes. Presence of these genes did not influence bacteria invasion in the liver and spleen of the chickens ten days after infection, although the presence of more than one fimbrial gene can be related to caecal colonization.A distribuição dos genes de virulência sefC, pefA e spvC foi investigada em 110 amostras de Salmonella Enteritidis pertencentes ao fagotipo 4 através da reação em cadeia da polimerase. A influência destes genes na colonização do ceco e invasão do fígado e baço em pintinhos de um dia de idade foi avaliada. Oito amostras foram negativas para o gene spvC, três para o gene pefA e uma amostra para o gene sefC. Estes resultados permitiram a classificação das amostras em quatro genótipos. A presença destes genes não influenciou a invasão da bactéria no fígado e baço das aves dez dias após a infecção, entretanto, a presença de mais de um gene fimbrial pode ter relação com a colonização cecal.

  19. Effects of L-arabinose efflux on λ Red recombination-mediated gene knockout in multiple-antimicrobial-resistant Salmonella enterica serovar Choleraesuis.

    Science.gov (United States)

    Liao, Shi-Wei; Lee, Jen-Jie; Ptak, Christopher P; Wu, Ying-Chen; Hsuan, Shih-Ling; Kuo, Chih-Jung; Chen, Ter-Hsin

    2017-10-03

    In this study, six swine-derived multiple-antimicrobial-resistant (MAR) strains of Salmonella Choleraesuis (S. Choleraesuis) were demonstrated to possess higher efflux pump activity than the wild-type (WT). L-Arabinose, a common inducer for gene expression, modulated S. Choleraesuis efflux pump activity in a dose-dependent manner. At low L-arabinose concentrations, increasing L-arabinose led to a corresponding increase in fluorophore efflux, while at higher L-arabinose concentrations, increasing L-arabinose decreased fluorophore efflux activity. The WT S. Choleraesuis that lacks TolC (ΔtolC), an efflux protein associated with bacterial antibiotic resistance and virulence, was demonstrated to possess a significantly reduced ability to extrude L-arabinose. Further, due to the rapid export of L-arabinose, an efficient method for recombination-mediated gene knockout, the L-arabinose-inducible bacteriophage λ Red recombinase system, has a reduced recombination frequency (~ 12.5%) in clinically isolated MAR Salmonella strains. An increased recombination frequency (up to 60%) can be achieved using a higher concentration of L-arabinose (fivefold) for genetic manipulation and functional analysis for MAR Salmonella using the λ Red system. The study suggests that L-arabinose serves not only as an inducer of the TolC-dependent efflux system but also acts as a competitive substrate of the efflux system. In addition, understanding the TolC-dependent efflux of L-arabinose should facilitate the optimization of L-arabinose induction in strains with high efflux activity.

  20. Gene Expression Response of Salmonella enterica Serotype Enteritidis Phage Type 8 to Subinhibitory Concentrations of the Plant-Derived Compounds Trans-Cinnamaldehyde and Eugenol

    Directory of Open Access Journals (Sweden)

    Anup Kollanoor Johny

    2017-09-01

    Full Text Available Background:Salmonella Enteritidis phage type 8 (PT8 is a major poultry-associated Salmonella strain implicated in foodborne outbreaks in the United States. We previously reported that two plant-derived compounds generally recognized as safe (GRAS, trans-cinnamaldehyde (TC, and eugenol (EG, significantly reduced S. Enteritidis colonization in broiler and layer chickens. To elucidate potential PT8 genes affected by TC and EG during colonization, a whole-genome microarray analysis of the bacterium treated with TC and EG was conducted.Results:S. Enteritidis PT8 was grown in Luria-Bertani broth at 37°C to an OD600 of ~0.5. Subinhibitory concentrations (SICs; concentration that does not inhibit bacterial growth of TC (0.01%; 0.75 mM or EG (0.04%; 2.46 mM were then added to the culture. S. Enteritidis PT8 RNA was extracted before and 30 min after TC or EG addition. Labeled cDNA from three replicate experiments was subsequently hybridized to a microarray of over 99% of S. Enteritidis PT4 genes, and the hybridization signals were quantified. The plant-derived compounds down-regulated (P < 0.005 expression of S. Enteritidis PT8 genes involved in flagellar motility, regulation of the Salmonella Pathogenicity Island 1, and invasion of intestinal epithelial cells. TC and EG also suppressed transcription of genes encoding multiple transport systems and outer membrane proteins. Moreover, several metabolic and biosynthetic pathways in the pathogen were down-regulated during exposure to the plant-derived compounds. Both TC and EG stimulated the transcription of heat shock genes, such as dnaK, dnaJ, ibpB, and ibpA in S. Enteritidis PT8 (P < 0.005. The results obtained from microarray were validated using a quantitative real-time PCR.Conclusion: The plant-derived compounds TC and EG exert antimicrobial effects on S. Enteritidis PT8 by affecting multiple genes, including those associated with virulence, colonization, cell membrane composition, and transport

  1. Distribution of prophages and SGI-1 antibiotic-resistance genes among different Salmonella enterica serovar Typhimurium isolates

    NARCIS (Netherlands)

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

    2006-01-01

    Recently, the authors identified Salmonella enterica serovar Typhimurium (S. Typhimurium) definitive type (DT)104-specific sequences of mainly prophage origin by genomic subtractive hybridization. In the present study, the distribution of the prophages identified, ST104 and ST64B, and the novel

  2. MUTATIONAL AND TRANSCRIPTIONAL RESPONSE OF SALMONELLA TO MX: CORRELATION OF MUTATIONAL DOSE RESPONSE TO CHANGES IN GENE EXPRESSION

    Science.gov (United States)

    We measured the mutational and transcriptional response of Salmonella TA100 to 3 concentrations of the drinking water mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy2(5H)-furanone (MX). The mutagenicity of MX in strain TA100 was evaluated in a 30min suspension assay, and the mutage...

  3. Development of a DNA microarray to detect antimicrobial resistance genes identified in the National Center for Biotechnology Information database.

    Science.gov (United States)

    Frye, Jonathan G; Lindsey, Rebecca L; Rondeau, Gaelle; Porwollik, Steffen; Long, Fred; McClelland, Michael; Jackson, Charlene R; Englen, Mark D; Meinersmann, Richard J; Berrang, Mark E; Davis, Johnnie A; Barrett, John B; Turpin, Jennifer B; Thitaram, Sutawee N; Fedorka-Cray, Paula J

    2010-03-01

    To understand the mechanisms and epidemiology of antimicrobial resistance (AR), the genetic elements responsible must be identified. Due to the myriad of possible genes, a high-density genotyping technique is needed for initial screening. To achieve this, AR genes in the National Center for Biotechnology Information GenBank database were identified by their annotations and compiled into a nonredundant list of 775 genes. A DNA microarray was constructed of 70mer oligonucelotide probes designed to detect these genes encoding resistances to aminoglycosides, beta-lactams, chloramphenicols, glycopeptides, heavy metals, lincosamides, macrolides, metronidazoles, polyketides, quaternary ammonium compounds, streptogramins, sulfonamides, tetracyclines, and trimethoprims as well as resistance transfer genes. The microarray was validated with two fully sequenced control strains of Salmonella enterica: Typhimurium LT2 (sensitive) and Typhi CT18 (multidrug resistance [MDR]). All resistance genes encoded on the MDR plasmid, pHCM1, harbored by CT18 were detected in that strain, whereas no resistance genes were detected in LT2. The microarray was also tested with a variety of bacteria, including MDR Salmonella enterica serovars, Escherichia coli, Campylobacter spp., Enterococcus spp., methicillin-resistant Staphylococcus aureus, Listeria spp., and Clostridium difficile. The results presented here demonstrate that a microarray can be designed to detect virtually all AR genes found in the National Center for Biotechnology Information database, thus reducing the subsequent assays necessary to identify specific resistance gene alleles.

  4. Effect of Plant Derived Antimicrobials on Salmonella Enteritidis Adhesion to and Invasion of Primary Chicken Oviduct Epithelial Cells in vitro and Virulence Gene Expression

    Directory of Open Access Journals (Sweden)

    Kumar Venkitanarayanan

    2013-05-01

    Full Text Available Salmonella Enteritidis (SE is a major foodborne pathogen in the United States and one of the most frequently reported Salmonella serotypes globally. Eggs are the most common food product associated with SE infections in humans. The pathogen colonizes the intestinal tract in layers, and migrates to reproductive organs systemically. Since adhesion to and invasion of chicken oviduct epithelial cells (COEC is critical for SE colonization in reproductive tract, reducing these virulence factors could potentially decrease egg yolk contamination. This study investigated the efficacy of sub-inhibitory concentrations of three plant-derived antimicrobials (PDAs, namely carvacrol, thymol and eugenol in reducing SE adhesion to and invasion of COEC, and survival in chicken macrophages. In addition, the effect of PDAs on SE genes critical for oviduct colonization and macrophage survival was determined using real-time quantitative PCR (RT-qPCR. All PDAs significantly reduced SE adhesion to and invasion of COEC (p < 0.001. The PDAs, except thymol consistently decreased SE survival in macrophages (p < 0.001. RT-qPCR results revealed down-regulation in the expression of genes involved in SE colonization and macrophage survival (p < 0.001. The results indicate that PDAs could potentially be used to control SE colonization in chicken reproductive tract; however, in vivo studies validating these results are warranted.

  5. Differentiation of UK endemic strains of Salmonella enterica serovar Newport from epidemic North American strains by PCR detection of a truncated bapA chromosomal gene.

    Science.gov (United States)

    Horton, R A; Card, R; Randall, L P; Teale, C J

    2016-02-01

    The aim of this study was to develop a PCR test to detect chromosomal differences between epidemic multidrug resistant (epi-MDR) strains of Salmonella enterica serovar Newport (S. Newport) and non-epi-MDR strains of S. Newport that are endemic to the United Kingdom (UK). Sequence analysis of the biofilm-associated protein A gene (bapA) showed that epi-MDR strains of S. Newport from the United States of America (USA) had a deletion of 309 bp, which was not present in non-epi-MDR strains of S. Newport from the UK. A PCR test was developed using primers designed to target this difference and was applied to a panel of S. Newport isolates comprising of strains from the UK (n=20, non-epi-MDR), from the USA (n=10, epi-MDR) and from Canada (n=7). A second panel of isolates (n=73) was used to assess the test specificity, and these isolates consisted of non-Newport Salmonella serovars (n=25), and other epidemic serovars (n=48). Epi-MDR S. Newport isolates produced a characteristic 505 bp amplicon, whereas non-epi-MDR S. Newport isolates produced an 814 bp amplicon. The bapA PCR has potential to discriminate between these S. Newport strains irrespective of their carrying resistance genes. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

  6. Validation of DNA and RNA real-time assays for food analysis using the hilA gene of Salmonella enterica serovars.

    Science.gov (United States)

    McCabe, Evonne M; Burgess, Catherine M; Walsh, Des; O'Regan, Edel; McGuinness, Sheila; Barry, Thomas; Fanning, Séamus; Duffy, Geraldine

    2011-01-01

    In Europe, alternative methods for the detection of food-borne pathogens can be used instead of the standard ISO/CEN reference protocol, if validated according to the protocol outlined in ISO 16140, 2003. In this study, the performance of two novel methods for the detection of Salmonella sp. using real-time PCR technology in tandem with an adapted two-step enrichment protocol were assessed and validated against a reference culture method, ISO 6579, 2004. The DNA and RNA real-time PCR assays amplified a 270 bp region of the hilA gene of Salmonella enterica serovars, and incorporated an internal amplification control (IAC) which was co-amplified with the hilA gene to monitor potential PCR inhibitors and ensure successful amplification. The inclusivity and exclusivity of the hilA primer set was examined for both the DNA and RNA methods and detected the 30 S. enterica serovars but not the 30 non-salmonellae strains. The inoculation of meat carcass swabs with five different S. enterica serovars at five different inocula, indicated both PCR methods were able to detect between 1 and 10 CFU per carcass swab. The real-time DNA PCR assay performed as well as the traditional cultural method in detecting Salmonella sp. in artificially contaminated salad, chocolate, fish and cheese samples. The relative accuracy, relative sensitivity and relative specificity of the DNA PCR real-time method were determined to be 98.5, 98.1 and 100%, respectively. The DNA method was further validated in a collaborative inter-laboratory trial according to ISO 16140, 2003. The validated methods provide an accurate means for the rapid detection and tracking of S. enterica serovars giving equivalent results to the standard method within three days, thus providing an alternative testing method to the reference microbiological method. The real-time PCR methodology not only offers significant time-saving advantages compared to traditional methods, it can also be applied to a wide range of samples types

  7. Conservation of Salmonella infection mechanisms in plants and animals.

    Science.gov (United States)

    Schikora, Adam; Virlogeux-Payant, Isabelle; Bueso, Eduardo; Garcia, Ana V; Nilau, Theodora; Charrier, Amélie; Pelletier, Sandra; Menanteau, Pierrette; Baccarini, Manuela; Velge, Philippe; Hirt, Heribert

    2011-01-01

    Salmonella virulence in animals depends on effectors injected by Type III Secretion Systems (T3SSs). In this report we demonstrate that Salmonella mutants that are unable to deliver effectors are also compromised in infection of Arabidopsis thaliana plants. Transcriptome analysis revealed that in contrast to wild type bacteria, T3SS mutants of Salmonella are compromised in suppressing highly conserved Arabidopsis genes that play a prominent role during Salmonella infection of animals. We also found that Salmonella originating from infected plants are equally virulent for human cells and mice. These results indicate a high degree of conservation in the defense and infection mechanism of animal and plant hosts during Salmonella infection.

  8. Antimicrobial susceptibility and serovars of Salmonella from chickens and humans in Ibadan, Nigeria

    DEFF Research Database (Denmark)

    Fashae, K; Ogunsola, F; Aarestrup, Frank Møller

    2010-01-01

    BACKGROUND: This study determines the prevalence and antibiotic resistance of Salmonella serovars from humans and chickens in Ibadan, Nigeria, in 2004-2007. METHODOLOGY: A total of 991 blood samples were collected from patients in 2004 to 2005 and 641 fecal samples were collected from poultry farms...... in 2007. All Salmonella isolates were serotyped and tested for antimicrobial susceptibility. RESULTS: Thirty-nine (4%) Salmonella isolates were obtained from human blood and 70 (11%) from chicken fecal samples. The human isolates revealed nine different serovars; 82% were non-typhoidal Salmonella and 18......% were (S. Typhi). The majority of serovars from humans were S. Enteritidis (33%), S. Dublin (18%), and S. Typhimurium (18%). Resistance to chloramphenicol, sulfamethoxazole, trimethoprim, and ampicillin ranged from 36% to 59% for the human isolates. Eight different serovars were obtained from chickens...

  9. The contribution of genes required for anaerobic respiration to the virulence of Salmonella enterica serovar Gallinarum for chickens.

    Science.gov (United States)

    Paiva, J B; Penha Filho, R A C; Pereira, E A; Lemos, M V F; Barrow, P A; Lovell, M A; Berchieri, A

    2009-10-01

    Salmonella enterica serovar Gallinarum (SG) is an intracellular pathogen of chickens. To survive, to invade and to multiply in the intestinal tract and intracellularly it depends on its ability to produce energy in anaerobic conditions. The fumarate reductase (frdABCD), dimethyl sulfoxide (DMSO)-trimethylamine N-oxide (TMAO) reductase (dmsABC), and nitrate reductase (narGHIJ) operons in Salmonella Typhimurium (STM) encode enzymes involved in anaerobic respiration to the electron acceptors fumarate, DMSO, TMAO, and nitrate, respectively. They are regulated in response to nitrate and oxygen availability and changes in cell growth rate. In this study mortality rates of chickens challenged with mutants of Salmonella Gallinarum, which were defective in utilising anaerobic electron acceptors, were assessed in comparison to group of bird challenged with wild strain. The greatest degree of attenuation was observed with mutations affecting nitrate reductase (napA, narG) with additional attenuations induced by a mutation affecting fumarate reductase (frdA) and a double mutant (dmsA torC) affecting DMSO and TMAO reductase.

  10. Evaluation of the GeneQuence DNA hybridization method in conjunction with 24-hour enrichment protocols for detection of Salmonella spp. in select foods: collaborative study.

    Science.gov (United States)

    Mozola, Mark A; Peng, Xuan; Wendorf, Michael

    2007-01-01

    A multilaboratory study was conducted to compare performance of the GeneQuence DNA hybridization (DNAH) method incorporating new 24 h enrichment protocols and reference culture procedures for detection of Salmonella spp. in select foods. Six food types (raw ground turkey, raw ground beef, dried whole egg, milk chocolate, walnuts, and dry pet food) were tested by the DNAH method and by the culture methods of either the U.S. Department of Agriculture-Food Safety and Inspection Service (USDA-FSIS) or the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA/BAM). Fifteen laboratories participated in the study. Four of the foods tested (raw ground turkey, dried whole egg, milk chocolate, and dry pet food), showed no statistically significant differences in performance between the DNAH method and the reference procedure as determined by Chi square analysis. Sensitivity rates for the DNAH method ranged from 92 to 100%. The DNAH method, with the specific enrichment protocol evaluated, was found to be ineffective for detection of Salmonella spp. in walnuts. For raw ground beef, results from one trial showed a statistically significant difference in performance, with more positives obtained by the reference method. However, evidence suggests that the difference in the number of positives was likely due to lack of homogeneity of the test samples rather than to DNAH method performance.

  11. Colostrum whey down-regulates the expression of early and late inflammatory response genes induced by Escherichia coli and Salmonella enterica Typhimurium components in intestinal epithelial cells.

    Science.gov (United States)

    Blais, M; Fortier, M; Pouliot, Y; Gauthier, S F; Boutin, Y; Asselin, C; Lessard, M

    2015-01-28

    Pathogenic invasion by Escherichia coli and Salmonellae remains a constant threat to the integrity of the intestinal epithelium and can rapidly induce inflammatory responses. At birth, colostrum consumption exerts numerous beneficial effects on the properties of intestinal epithelial cells and protects the gastrointestinal tract of newborns from pathogenic invasion. The present study aimed to investigate the effect of colostrum on the early and late inflammatory responses induced by pathogens. The short-term (2 h) and long-term (24 h) effects of exposure to heat-killed (HK) E. coli and Salmonella enterica Typhimurium on gene expression in the porcine intestinal epithelial cell (IPEC-J2) model were first evaluated by microarray and quantitative PCR analyses. Luciferase assays were performed using a NF-κB-luc reporter construct to investigate the effect of colostrum whey treatment on the activation of NF-κB induced by HK bacteria. Luciferase assays were also performed using NF-κB-luc, IL-8-luc and IL-6-luc reporter constructs in human colon adenocarcinoma Caco-2/15 cells exposed to dose-response stimulations with HK bacteria and colostrum whey. Bovine colostrum whey treatment decreased the expression of early and late inflammatory genes induced by HK bacteria in IPEC-J2, as well as the transcriptional activation of NF-κB-luc induced by HK bacteria. Unlike that with colostrum whey, treatment with other milk fractions failed to decrease the activation of NF-κB-luc induced by HK bacteria. Lastly, the reduction of the HK bacteria-induced activation of NF-κB-luc, IL-8-luc and IL-6-luc by colostrum whey was dose dependent. The results of the present study indicate that bovine colostrum may protect and preserve the integrity of the intestinal mucosal barrier in the host by controlling the expression levels of early and late inflammatory genes following invasion by enteric pathogens.

  12. Antimicrobial activity of the bioactive components of essential oils from Pakistani spices against Salmonella and other multi-drug resistant bacteria

    Science.gov (United States)

    2013-01-01

    Background The main objective of this study was the phytochemical characterization of four indigenous essential oils obtained from spices and their antibacterial activities against the multidrug resistant clinical and soil isolates prevalent in Pakistan, and ATCC reference strains. Methods Chemical composition of essential oils from four Pakistani spices cumin (Cuminum cyminum), cinnamon (Cinnamomum verum), cardamom (Amomum subulatum) and clove (Syzygium aromaticum) were analyzed on GC/MS. Their antibacterial activities were investigated by minimum inhibitory concentration (MIC) and Thin-Layer Chromatography-Bioautographic (TLC-Bioautographic) assays against pathogenic strains Salmonella typhi (D1 Vi-positive), Salmonella typhi (G7 Vi-negative), Salmonella paratyphi A, Escherichia coli (SS1), Staphylococcus aureus, Pseudomonas fluorescens and Bacillus licheniformis (ATCC 14580). The data were statistically analyzed by using Analysis of Variance (ANOVA) and Least Significant Difference (LSD) method to find out significant relationship of essential oils biological activities at p spices can be pursued against multidrug resistant bacteria. PMID:24119438

  13. Gene Expression Analysis of Toll-like Receptor Pathways in Heterophils from Genetic Chicken Lines That Differ In Their Susceptibility to Salmonella enteritidis

    Directory of Open Access Journals (Sweden)

    Michael eKogut

    2012-07-01

    Full Text Available Previously conducted studies using two chicken lines (A and B show that line A birds have increased resistance to a number of bacterial and protozoan challenges and that heterophils isolated from line A birds are functionally more responsive. Furthermore, when stimulated with toll-like receptor (TLR agonists, heterophils from line A expressed a totally different cytokine and chemokine mRNA expression pattern than heterophils from line B. A large-scale gene expression profile using an Agilent 44K microarray on heterophils isolated from line A and line B also revealed significantly differential expression in many immune-related genes following Salmonella enteritidis (SE stimulation, which included genes involved in the TLR pathway. Therefore, we hypothesize the differences between the lines result from distinctive TLR pathway signaling cascades that mediate heterophil function and, thus, innate immune responsiveness to SE. Using quantitative RT-PCR on mRNA from heterophils isolated from control and SE-stimulated heterophils of each line, we profiled the expression of all chicken homologous genes identified in a reference TLR pathway. Several differentially expressed genes found were involved in the TLR-induced My88-dependent pathway, showing higher gene expression in line A than line B heterophils following SE stimulation. These genes included the toll-like receptor genes TLR4, TLR15, TLR21, MD2, the adaptor proteins toll-interleukin 1 receptor domain containing adaptor protein (TIRAP, Tumor necrosis factor-receptor associated factor 3 (TRAF3, the IκB kinases TGF-β-activating kinase 1 (TAK1, IKKε and IKKα, the transcription factors NFkB2 and interferon regulatory factor 7 (IRF7, phosphoinositol-3 kinase (PI-3K, and the mitogen-activated protein kinase (MAPK p38. These results indicate that higher expression of TLR signaling activation of both MyD88-dependent and TRIF-dependent pathways are more beneficial to avian heterophil-mediated innate

  14. Flagella overexpression attenuates Salmonella pathogenesis.

    Directory of Open Access Journals (Sweden)

    Xinghong Yang

    Full Text Available Flagella are cell surface appendages involved in a number of bacterial behaviors, such as motility, biofilm formation, and chemotaxis. Despite these important functions, flagella can pose a liability to a bacterium when serving as potent immunogens resulting in the stimulation of the innate and adaptive immune systems. Previous work showing appendage overexpression, referred to as attenuating gene expression (AGE, was found to enfeeble wild-type Salmonella. Thus, this approach was adapted to discern whether flagella overexpression could induce similar attenuation. To test its feasibility, flagellar filament subunit FliC and flagellar regulon master regulator FlhDC were overexpressed in Salmonella enterica serovar Typhimurium wild-type strain H71. The results show that the expression of either FliC or FlhDC alone, and co-expression of the two, significantly attenuates Salmonella. The flagellated bacilli were unable to replicate within macrophages and thus were not lethal to mice. In-depth investigation suggests that flagellum-mediated AGE was due to the disruptive effects of flagella on the bacterial membrane, resulting in heightened susceptibilities to hydrogen peroxide and bile. Furthermore, flagellum-attenuated Salmonella elicited elevated immune responses to Salmonella presumably via FliC's adjuvant effect and conferred robust protection against wild-type Salmonella challenge.

  15. Flagella Overexpression Attenuates Salmonella Pathogenesis

    Science.gov (United States)

    Yang, Xinghong; Thornburg, Theresa; Suo, Zhiyong; Jun, SangMu; Robison, Amanda; Li, Jinquan; Lim, Timothy; Cao, Ling; Hoyt, Teri; Avci, Recep; Pascual, David W.

    2012-01-01

    Flagella are cell surface appendages involved in a number of bacterial behaviors, such as motility, biofilm formation, and chemotaxis. Despite these important functions, flagella can pose a liability to a bacterium when serving as potent immunogens resulting in the stimulation of the innate and adaptive immune systems. Previous work showing appendage overexpression, referred to as attenuating gene expression (AGE), was found to enfeeble wild-type Salmonella. Thus, this approach was adapted to discern whether flagella overexpression could induce similar attenuation. To test its feasibility, flagellar filament subunit FliC and flagellar regulon master regulator FlhDC were overexpressed in Salmonella enterica serovar Typhimurium wild-type strain H71. The results show that the expression of either FliC or FlhDC alone, and co-expression of the two, significantly attenuates Salmonella. The flagellated bacilli were unable to replicate within macrophages and thus were not lethal to mice. In-depth investigation suggests that flagellum-mediated AGE was due to the disruptive effects of flagella on the bacterial membrane, resulting in heightened susceptibilities to hydrogen peroxide and bile. Furthermore, flagellum-attenuated Salmonella elicited elevated immune responses to Salmonella presumably via FliC’s adjuvant effect and conferred robust protection against wild-type Salmonella challenge. PMID:23056473

  16. A cross sectional study on antibiotic resistance pattern of Salmonella typhi clinical isolates from Bangladesh

    Directory of Open Access Journals (Sweden)

    Adnan Mannan

    2014-04-01

    Conclusions: Although the population density of Dhaka region is markedly higher than Rajshahi and Chittagong regions, no significant difference in resistance pattern was found. The rate of multidrug resistance is a matter of concern. Physicians should reconsider before prescribing nalidixic acid and cefixime. Further molecular study is needed to reveal the genomic and proteomic basis of resistance.

  17. Characterization of epidemic IncI1-Iγ plasmids harboring ambler class A and C genes in Escherichia coli and Salmonella enterica from animals and humans.

    Science.gov (United States)

    Smith, Hilde; Bossers, Alex; Harders, Frank; Wu, Guanghui; Woodford, Neil; Schwarz, Stefan; Guerra, Beatriz; Rodríguez, Irene; van Essen-Zandbergen, Alieda; Brouwer, Michael; Mevius, Dik

    2015-09-01

    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 β-lactamase (ESBL) or AmpC β-lactamase genes were compared using plasmid multilocus sequence typing (pMLST). Thirty-two of these plasmids belonging to different pMLST types were sequenced using Roche 454 and Illumina platforms. Epidemic IncI1-Iγ plasmids could be assigned to various dominant clades, whereas rarely detected plasmids clustered together as a distinct clade. Similar phylogenetic trees were obtained using only the plasmid backbone sequences, showing that the differences observed between the plasmids belonging to distinct clades resulted mainly from differences between their backbone sequences. Plasmids belonging to the various clades differed particularly in the presence/absence of genes encoding partitioning and addiction systems, which contribute to stable inheritance during cell division and plasmid maintenance. Despite this, plasmids belonging to the various phylogenetic clades also showed marked resistance gene associations, indicating the circulation of successful plasmid-gene combinations. The variation in traY and excA genes found in IncI1-Iγ plasmids is conserved within pMLST sequence types and plays a role in incompatibility, although functional study is needed to elucidate the role of these genes in plasmid epidemiology. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. Antibiotic resistance of prevalent Salmonella and Shigella strains in northwest Ethiopia.

    Science.gov (United States)

    Aseffa, A; Gedlu, E; Asmelash, T

    1997-11-01

    Salmonella and Shigella strains were isolated prospectively from in- and outpatient specimens of the Gondar College teaching hospital over a two year period, from June 1994 to May 1996. Of 7993 miscellaneous specimens cultured, 80 yielded Salmonella and 147 Shigella. Serogroup B dominated among the salmonella, accounting for 61% of isolates, followed by S. typhi (21%). S. flexneri (58.5%) and S. dysenteriae (36.7%) were the most frequently isolated species among the Shigella. Results of sensitivity testing to five commonly used antibiotics in the area: ampicillin, chloramphenicol, gentamicin, tetracycline and cotrimoxazole, showed multiple drug resistance among the isolates of both salmonella and shigella. Only 4% of shigella were sensitive to all five antibiotics screened for in the study. Overall sensitive of shigella isolates to individual antibiotics was 8.8% to tetracycline, 10% to ampicillin, 28% to co-trimoxazole and 98% to gentamicin. No resistance was observed to nalidixic acid among 108 shigella strains tested for this antibiotic. The continued sensitivity of S. typhi to chloramphenicol, cotrimoxazole, ampicillin and the other commonly used antibiotics is noted. It is recommended that nalidixic acid be introduced into the area under strict regulation for treatment of severe cases of shigella dysentery, an illness which is highly endemic in the region and the cause of occasional epidemics with high mortality.

  19. 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.

  20. Animal salmonelloses: a brief review of “host adaptation and host specificity” of Salmonella spp.

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    Grammato Evangelopoulou

    2013-07-01

    Full Text Available Salmonella enterica, the most pathogenic species of the genusSalmonella, includes more than 2,500 serovars, many of which are of great veterinary and medical significance. The emergence of food-borne pathogens, such as Salmonella spp., has increased knowledge about the mechanisms helping microorganisms to persist and spread within new host populations. It has also increased information about the properties they acquire for adapting in the biological environment of a new host. Thedifferences observed between serovars in their host preference and clinical manifestations are referred to as “serovar-host specificity” or “serovar-host adaptation”. The genus Salmonella, highly adaptive to vertebrate hosts, has many pathogenic serovars showing host specificity. Serovar Salmonella Typhi, causing disease to man and higher primates, is a good example of host specificity. Thus, understanding the mechanisms that Salmonella serovars use to overcome animal species' barriers or adapt to new hosts is also important for understanding the origins of any other infectious diseases or the emergence of new pathogens. In addition, molecular methods used to study the virulence determinants of Salmonella serovars, could also be used to model ways of studying the virulence determinants used by bacteria in general, when causing disease to a specific animal species

  1. Elevated temperature technique for the isolation of salmonellas from sewage and human faeces.

    Science.gov (United States)

    Nabbut, N H

    1973-03-01

    Modified Moore's swabs, placed in sewers for 5 days, were used to concentrate salmonellas from sewage. Duplicate cultures of swab strips in selenite broth were incubated at 41 and 37 degrees C. respectively. Salmonella organisms were recovered consistently from the swabs when the enrichment broths were incubated at 41 degrees C. However, when equal portions of the same swabs were incubated at 37 degrees C., only 22% of them yielded Salmonella organisms. These results indicate an advantage in incubating the selenite broths at 41 degrees C. rather than 37 degrees C. in attempting to isolate salmonellas from sewage.One hundred and fifty faecal samples were examined for salmonellas by culture in selenite broths incubated at 41 and 37 degrees C. Twelve (8%) samples were positive at 41 degrees C. compared to only 10 (6.7%) positive samples at 37 degrees C. This difference is not statistically significant to indicate an advantage of the elevated-temperature of incubation over the conventional temperature in attempting to isolate salmonellas from human faeces. Moreover, results of the recovery rates of S. paratyphi B, S. typhi, and S. typhimurium indicate that an incubation temperature of 37 degrees C. is more appropriate for recovering salmonellas from artificially infected faecal samples than an incubation temperature of 41 degrees C. This stresses the inability of laboratory studies to mimic conditions in nature.

  2. Salmonella in effluent from sewage treatment plants, wastepipe of butcher's shops and surface water in Walcheren.

    Science.gov (United States)

    Kampelmacher, E H; van Noorle Jansen, L M

    1976-07-01

    In the frame of the "Walcheren-project" in which the epidemiology of salmonellosis is studied in a certain area, effluent from sewage treatment plants, wastepipe's of butcher's shops and surface waters, which receive the effluent were studied for the presence of salmonellae. From 160 samples of effluent 150 (94%) contained salmonellae. The most common serotype was S. typhi murium (35%) followed by S. panama and S. infantis. 14 butcher's shops' wastepipes were sampled 54 times. 14 (26%) times salmonellae were found, but only twice was the type isolated from the butcher's shop the same as found in the effluent on the same day. With regard to the presence of salmonellae in surface waters receiving effluent it was shown that from the immediate vicinity of the plant to 250 m downstream from the site of drainage of effluent the number of salmonellae per 100 ml remains almost constant. After 1.5-4 kilometers Salmonella could not be isolated from any of the samples examined. The results underline the hypothesis that salmonellae multiply in the sewage system and/or plant. The spread of samonellae by effluent seems to be limited to the plant itself and of the nearest vicinity. Proposals are brought forward to interupt contamination cycles by decontamination measures.

  3. Suppressive effects of retinoids, carotenoids and antioxidant vitamins on heterocyclic amine-induced umu C gene expression in Salmonella typhimurium (TA 1535/pSK 1002).

    Science.gov (United States)

    Okai, Y; Higashi-Okai, K; Nakamura, S; Yano, Y; Otani, S

    1996-06-12

    Effects of retinoids, carotenoids and antioxidant vitamins were studied by mutagen-induced umu C gene expression system in Salmonella typhimurium (TA 1535/pSK 1002). Retinol (vitamin A), retinol acetate and retinoic acid showed remarkable inhibitory activities, whereas retinol palmitate exhibited significant but weak activity for umu C gene expression in tester bacteria induced by 3-amino-3,4-dimethyl-5H-pyrido[4.3-b]indol (Trp-P-1) in the presence of hepatic metabolizing enzymes (S9 mixture). Carotenoids having provitamin A activity (beta-carotene and canthaxanthin) exhibited moderate suppressive effects on the same experimental system. The ranks of suppressive activities were retinol > retinol acetate > retinoic acid > canthaxanthin > beta-carotene > retinol palmitate and their doses for inhibition by 50% (ID50) were estimated to be 1.2 x 10(-7), 3.0 x 10(-7), 5.4 x 10(-7), 1.5 x 10(-6), 4.0 x 10(-5) and 6.0 x 10(-5) M, respectively. However, they did not cause significant inhibition on umu C gene expression induced by direct-acting mutagen (adriamycin or mitomycin C) in the absence of S9 mixture. Inhibition of umu gene expression appears to be due to inhibition of P450-mediated metabolic activation of the heterocyclic amine Trp-P-1. Ascorbic acid (vitamin C) showed weak but significant suppressive activity at high-dose concentrations (3 x 10(-6) - 10(-4)M). However, alpha-tocopherol did not exhibit significant suppression at all dose concentrations. The significance of the experimental results is discussed from the viewpoint of the chemoprevention against genotoxicity associated with carcinogenesis.

  4. Genes ycfR, sirA and yigG contribute to the surface attachment of Salmonella enterica Typhimurium and Saintpaul to fresh produce.

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    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.

  5. Gene expression response of Salmonella enterica serotype Enteritidis phage type 8 to the subinhibitory concentrations of the plant-derived compounds,trans-cinnamaldehyde,and eugenol

    Science.gov (United States)

    Background: Salmonella Enteritidis phage type 8 (PT8) is a major poultry-associated Salmonella strain implicated in foodborne outbreaks in the United States. We previously reported that two GRAS-status, plant-derived compounds, trans-cinnamaldehyde (TC) and eugenol (EG) significantly reduced S. Ent...

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

    Science.gov (United States)

    Salmonella Enteritidis is the world’s leading cause of food borne salmonellosis and illness in people is linked strongly to its contamination of eggs produced by otherwise healthy appearing hens. Salmonella Enteritidis is noted for generating exceptional strain heterogeneity despite having a clonal ...

  7. 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. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. A Rapid and Sensitive Diagnosis of Typhoid Fever Based On Nested PCR-Voltammetric DNA Biosensor Using Flagellin Gene Fragment

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    Yeni Wahyuni Hartati

    2016-03-01

    Full Text Available Typhoid fever caused by Salmonella typhi is an important issue for public health in the world. Laboratory methods for rapid and sensitive diagnosis are very important for disease management. The purpose of this study was to determine the performance of nested PCR–voltammetric DNA biosensor using flagellin gene (fla of S. typhi as a marker. The differential pulse voltammetry using pencil graphite electrode was applied to measure the guanine oxidation signal of probes vs synthetic target stDNA and probes vs fla PCR product hybridizations. The probe DNA selectivity was examined by hybridized probes vs non-complementary sequence. The result showed that the first round nested PCR product can not be visualized by agarose electrophoresis, whereas using the voltammetric biosensor methods can be detected both for the first or second round nested PCR product. The average peak current of hybridized probe vs first and second round of PCR product was 2.32 and 1.47 μA respectively, at 0.9 V. Detection of the DNA sequences of the infectious diseases from PCR amplified real sample was also carried out using this voltammetric DNA biosensor methods.

  9. Microarray-based analysis of IncA/C Plasmid-Associated genes from multidrug-resistant Salmonella enterica

    Science.gov (United States)

    Bacteria plasmids are fragments of extra-chromosomal double stranded deoxyribonucleic acid (DNA) that can contain a variety of genes beneficial to the survival of the host bacteria. Classification and tracking of bacterial plasmids is valuable for the study of horizontal gene transfer of drug resis...

  10. Integrated Stress Responses in Salmonella

    Science.gov (United States)

    Shen, Shu; Fang, Ferric C.

    2011-01-01

    The foodborne gram-negative pathogen Salmonella must adapt to varied environmental conditions encountered within foods, the host gastrointestinal tract and the phagosomes of host macrophages. Adaptation is achieved through the coordinate regulation of gene expression in response to environmental signals such as temperature, pH, osmolarity, redox state, antimicrobial peptides, and nutrient deprivation. This review will examine mechanisms by which the integration of regulatory responses to a broad array of environmental signals can be achieved. First, in the most straightforward case, tandem promoters allow gene expression to respond to multiple signals. Second, versatile sensor proteins may respond to more than one environmental signal. Third, transcriptional silencing and counter-silencing as demonstrated by the H-NS paradigm provides a general mechanism for the convergence of multiple regulatory inputs. Fourth, signaling cascades allow gene activation by independent sensory elements. These mechanisms allow Salmonella to utilize common adaptive stress pathways in response to a diverse range of environmental conditions. PMID:21570144

  11. A previously uncharacterized gene stm0551 plays a repressive role in the regulation of type 1 fimbriae in Salmonella enterica serotype Typhimurium

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    Wang Ke-Chuan

    2012-06-01

    Full Text Available Abstract Background Salmonella enterica serotype Typhimurium produces surface-associated fimbriae that facilitate adherence of the bacteria to a variety of cells and tissues. Type 1 fimbriae with binding specificity to mannose residues are the most commonly found fimbrial type. In vitro, static-broth culture favors the growth of S. Typhimurium with type 1 fimbriae, whereas non-type 1 fimbriate bacteria are obtained by culture on solid-agar media. Previous studies demonstrated that the phenotypic expression of type 1 fimbriae is the result of the interaction and cooperation of the regulatory genes fimZ, fimY, fimW, and fimU within the fim gene cluster. Genome sequencing revealed a novel gene, stm0551, located between fimY and fimW that encodes an 11.4-kDa putative phosphodiesterase specific for the bacterial second messenger cyclic-diguanylate monophosphate (c-di-GMP. The role of stm0551 in the regulation of type 1 fimbriae in S. Typhimurium remains unclear. Results A stm0551-deleted stain constructed by allelic exchange constitutively produced type 1 fimbriae in both static-broth and solid-agar medium conditions. Quantative RT-PCR revealed that expression of the fimbrial major subunit gene, fimA, and one of the regulatory genes, fimZ, were comparably increased in the stm0551-deleted strain compared with those of the parental strain when grown on the solid-agar medium, a condition that normally inhibits expression of type 1 fimbriae. Following transformation with a plasmid possessing the coding sequence of stm0551, expression of fimA and fimZ decreased in the stm0551 mutant strain in both culture conditions, whereas transformation with the control vector pACYC184 relieved this repression. A purified STM0551 protein exhibited a phosphodiesterase activity in vitro while a point mutation in the putative EAL domain, substituting glutamic acid (E with alanine (A, of STM0551 or a FimY protein abolished this activity. Conclusions The finding that the

  12. Salmonella spp. and risk factors for the contamination of slaughtered cattle carcass from a slaughterhouse of Bahir Dar Town, Ethiopia

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    Gizachew Muluneh

    2015-02-01

    Full Text Available Objective: To assess the prevalence of Salmonella species and associated factors from cattle carcasses in abattoir of Bahir Dar, Ethiopia. Methods: A cross-sectional study was conducted among slaughtered cattle carcass samples from Bahir Dar abattoir from December 2012 to February 2013. A total of 300 carcasses from hind limb, abdomen and neck of one hundred cattle were collected and analyzed for the prevalence of Salmonella. An observation checklist was used to collect data on the risk factors for contamination of carcass. Results: Out of the total 300 carcass samples, 23 (7.6% were positive for Salmonella species. Eleven (47.8%, nine (39.1% and three (13.1% of the isolates were Salmonella group A, Salmonella arizonae and Salmonella typhi, respectively. Wearing garment during slaughtering, handwashing after separating intestinal content, washing of the knife before slaughtering, slaughtering on sanitized floor and carcass-washing during slaughter were important risk factors that have statistically significant association with the isolation rate of Salmonella in slaughtered cattle carcasses (P<0.05. The mean aerobic mesophilic count of cattle carcass surfaces was 5.39 log CFU/ cm2. Conclusions: Unlike other studies, Salmonella group A appears to be the most prevalent species in cattle carcasses in the present study. Among carcass sites, the highest contamination was recorded in the abdomen and the main source of contamination is human and intestinal contents. Therefore, there is a need to maintain proper hygienic practices in Bahir Dar abattoir during slaughtering.

  13. Subtyping Salmonella enterica serovar enteritidis isolates from different sources by using sequence typing based on virulence genes and clustered regularly interspaced short palindromic repeats (CRISPRs).

    Science.gov (United States)

    Liu, Fenyun; Kariyawasam, Subhashinie; Jayarao, Bhushan M; Barrangou, Rodolphe; Gerner-Smidt, Peter; Ribot, Efrain M; Knabel, Stephen J; Dudley, Edward G

    2011-07-01

    Salmonella enterica subsp. enterica serovar Enteritidis is a major cause of food-borne salmonellosis in the United States. Two major food vehicles for S. Enteritidis are contaminated eggs and chicken meat. Improved subtyping methods are needed to accurately track specific strains of S. Enteritidis related to human salmonellosis throughout the chicken and egg food system. A sequence typing scheme based on virulence genes (fimH and sseL) and clustered regularly interspaced short palindromic repeats (CRISPRs)-CRISPR-including multi-virulence-locus sequence typing (designated CRISPR-MVLST)-was used to characterize 35 human clinical isolates, 46 chicken isolates, 24 egg isolates, and 63 hen house environment isolates of S. Enteritidis. A total of 27 sequence types (STs) were identified among the 167 isolates. CRISPR-MVLST identified three persistent and predominate STs circulating among U.S. human clinical isolates and chicken, egg, and hen house environmental isolates in Pennsylvania, and an ST that was found only in eggs and humans. It also identified a potential environment-specific sequence type. Moreover, cluster analysis based on fimH and sseL identified a number of clusters, of which several were found in more than one outbreak, as well as 11 singletons. Further research is needed to determine if CRISPR-MVLST might help identify the ecological origins of S. Enteritidis strains that contaminate chickens and eggs.

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

    DEFF Research Database (Denmark)

    Herrero-Fresno, Ana; Espinel, Irene Cartas; Spiegelhauer, Malene Roed

    2018-01-01

    -pathogen interaction in S. Dublin, a mutant where this gene was deleted (S. Dublin ΔbstA) and a mutant which was further genetically complemented with bstA (S. Dublin 3246-C) were constructed and tested in models of in vitro and in vivo infection as well as during growth competition assays in M9 medium, Luria...... not affect either the amount of cytokines released by THP-1 macrophages or the cytotoxicity toward these cells. The histology of the livers and spleens of mice infected with the wild-type strain and the S. Dublin ΔbstA mutant revealed similar levels of inflammation between the two groups. The gene...

  15. Molecular methods for serovar determination of Salmonella.

    Science.gov (United States)

    Shi, Chunlei; Singh, Pranjal; Ranieri, Matthew Louis; Wiedmann, Martin; Moreno Switt, Andrea Isabel

    2015-01-01

    Salmonella is a diverse foodborne pathogen, which has more than 2600 recognized serovars. Classification of Salmonella isolates into serovars is essential for surveillance and epidemiological investigations; however, determination of Salmonella serovars, by traditional serotyping, has some important limitations (e.g. labor intensive, time consuming). To overcome these limitations, multiple methods have been investigated to develop molecular serotyping schemes. Currently, molecular methods to predict Salmonella serovars include (i) molecular subtyping methods (e.g. PFGE, MLST), (ii) classification using serovar-specific genomic markers and (iii) direct methods, which identify genes encoding antigens or biosynthesis of antigens used for serotyping. Here, we reviewed reported methodologies for Salmonella molecular serotyping and determined the "serovar-prediction accuracy", as the percentage of isolates for which the serovar was correctly classified by a given method. Serovar-prediction accuracy ranged from 0 to 100%, 51 to 100% and 33 to 100% for molecular subtyping, serovar-specific genomic markers and direct methods, respectively. Major limitations of available schemes are errors in predicting closely related serovars (e.g. Typhimurium and 4,5,12:i:-), and polyphyletic serovars (e.g. Newport, Saintpaul). The high diversity of Salmonella serovars represents a considerable challenge for molecular serotyping approaches. With the recent improvement in sequencing technologies, full genome sequencing could be developed into a promising molecular approach to serotype Salmonella.

  16. Salmonella Diagnosis and Treatment

    Science.gov (United States)

    ... FDA) USDA Food Safety and Inspection Service Follow Salmonella RSS Diagnosis and Treatment Recommend on Facebook Tweet Share Compartir How Can Salmonella Infections Be Diagnosed? Diagnosing salmonellosis requires testing a ...

  17. Salmonella in Swedish cattle

    OpenAIRE

    Ågren, Estelle

    2017-01-01

    In Sweden, all herds detected with salmonella are put under restrictions and measures aiming at eradication are required. The purpose of these studies was to provide a basis for decisions on how surveillance and control of salmonella in Swedish cattle can be made more cost-efficient. Results from a bulk milk screening were used to investigate seroprevalence of salmonella and to study associations between salmonella status and geographical location, local animal density, number of test pos...

  18. CHARACTERIZATION OF SALMONELLA SPECIES FROM WATER BODIES IN DAR-ES-SALAAM CITY, TANZANIA

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    Eliningaya Kweka

    2013-03-01

    Full Text Available Background: Water-borne diseases are the most common cause of illness and death among the poor population from developing countries. The majority of the people are inadequately aware that aquatic environment is a major source of salmonellosis. Dar es Salaam city is among the cities with most of its population live in squatter. Typhoid fever ranks second with 14.3% of all notifiable disease cases in the city. The city experience water scarcity which forces water wells and rivers to become the main sources of water for domestic use and livestock. This study therefore, characterized Salmonella strains from different water bodies of city as possible sources for enteric diseases endemicity. Methods: The Salmonella Chromogenic Agar (SC Agar and Kligler Iron Agar (KIA media were used for isolation and enumeration of the strains. The inoculated cultures were incubated at 370C for 24 hours. Salmonella colonies were confirmed by magenta colorations and hydrogen sulfide production on SC Agar and KIA Agar, respectively. The Analytical Profile Index 20 Enterobacteriaceae kit (API 20E kit was used to identify Salmonella species. Results: Based on the API 20E kit, the identified Salmonella species from different water bodies were Salmonella ser. paratyphi A (96.9%, Salmonella cholelaesuis spp choleraesuis (99.5% and Salmonella typhi (99.9%. Conclusion: This study shows that shallow wells and rivers which are mainly used by the city dwellers were highly contaminated with Salmonella and were more contaminated than deep wells and marine water bodies. This warrants further investigation on the disease mapping in the urban and peri-urban areas.

  19. 78 FR 42526 - Salmonella

    Science.gov (United States)

    2013-07-16

    ... HUMAN SERVICES Food and Drug Administration Salmonella Contamination of Dry Dog Food; Withdrawal of...) entitled ``Sec. 690.700 Salmonella Contamination of Dry Dog Food.'' This CPG is obsolete. DATES: The.... SUPPLEMENTARY INFORMATION: FDA issued the CGP entitled ``Sec. 690.700 Salmonella Contamination of Dry Dog Food...

  20. Flagella Overexpression Attenuates Salmonella Pathogenesis

    OpenAIRE

    Xinghong Yang; Theresa Thornburg; Zhiyong Suo; SangMu Jun; Amanda Robison; Jinquan Li; Timothy Lim; Ling Cao; Teri Hoyt; Recep Avci; Pascual, David W.

    2012-01-01

    Flagella are cell surface appendages involved in a number of bacterial behaviors, such as motility, biofilm formation, and chemotaxis. Despite these important functions, flagella can pose a liability to a bacterium when serving as potent immunogens resulting in the stimulation of the innate and adaptive immune systems. Previous work showing appendage overexpression, referred to as attenuating gene expression (AGE), was found to enfeeble wild-type Salmonella. Thus, this approach was adapted to...

  1. Whole Genome Epidemiological Typing of Salmonella

    DEFF Research Database (Denmark)

    Leekitcharoenphon, Pimlapas

    Salmonella is one of the most common foodborne pathogens worldwide. In the US alone, salmonellosis was estimated to cause 1.4 million cases effecting 17,000 hospitalization and almost 600 deaths each year. Particularly, Salmonella enterica is a common cause of minor and large food borne outbreaks...... used for typing is crucial for successful discrimination. The core genes or the genes that are conserved in all members of a genus or species are potentially good candidates for investigating genomic variation in phylogeny and epidemiology. A total of 2,882 core genes have been observed among 73....../absence of all genes across genomes, is similar to the consensus tree but with higher branching confidence value. 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. These core genes are useful for investigating molecular...

  2. Drug resistant Salmonella in broiler chicken sold at local market in ...

    African Journals Online (AJOL)

    This study was designed to isolate and identify Salmonella spp. from cloacal swabs of apparently healthy broiler chickens in Bangladesh. Salmonella was characterized culturally, biochemically and also via PCR method. Among 50 isolates, 16 were found to be positive for Salmonella. PCR using 16S rRNA gene primers ...

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

    Directory of Open Access Journals (Sweden)

    Harichandran D

    2017-03-01

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

  4. 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

    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...

  5. Design of a Comprehensive Biochemistry and Molecular Biology Experiment: Phase Variation Caused by Recombinational Regulation of Bacterial Gene Expression

    Science.gov (United States)

    Sheng, Xiumei; Xu, Shungao; Lu, Renyun; Isaac, Dadzie; Zhang, Xueyi; Zhang, Haifang; Wang, Huifang; Qiao, Zheng; Huang, Xinxiang

    2014-01-01

    Scientific experiments are indispensable parts of Biochemistry and Molecular Biology. In this study, a comprehensive Biochemistry and Molecular Biology experiment about "Salmonella enterica" serovar Typhi Flagellar phase variation has been designed. It consisted of three parts, namely, inducement of bacterial Flagellar phase variation,…

  6. Ultra-fast and sensitive detection of non-typhoidal Salmonella using microwave-accelerated metal-enhanced fluorescence ("MAMEF".

    Directory of Open Access Journals (Sweden)

    Sharon M Tennant

    Full Text Available Certain serovars of Salmonella enterica subsp. enterica cause invasive disease (e.g., enteric fever, bacteremia, septicemia, meningitis, etc. in humans and constitute a global public health problem. A rapid, sensitive diagnostic test is needed to allow prompt initiation of therapy in individual patients and for measuring disease burden at the population level. An innovative and promising new rapid diagnostic technique is microwave-accelerated metal-enhanced fluorescence (MAMEF. We have adapted this assay platform to detect the chromosomal oriC locus common to all Salmonella enterica subsp. enterica serovars. We have shown efficient lysis of biologically relevant concentrations of Salmonella spp. suspended in bacteriological media using microwave-induced lysis. Following lysis and DNA release, as little as 1 CFU of Salmonella in 1 ml of medium can be detected in <30 seconds. Furthermore the assay is sensitive and specific: it can detect oriC from Salmonella serovars Typhi, Paratyphi A, Paratyphi B, Paratyphi C, Typhimurium, Enteritidis and Choleraesuis but does not detect Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae or Acinetobacter baumanii. We have also performed preliminary experiments using a synthetic Salmonella oriC oligonucleotide suspended in whole human blood and observed rapid detection when the sample was diluted 1:1 with PBS. These pre-clinical data encourage progress to the next step to detect Salmonella in blood (and other ordinarily sterile, clinically relevant body fluids.

  7. Ultra-fast and sensitive detection of non-typhoidal Salmonella using microwave-accelerated metal-enhanced fluorescence ("MAMEF").

    Science.gov (United States)

    Tennant, Sharon M; Zhang, Yongxia; Galen, James E; Geddes, Chris D; Levine, Myron M

    2011-04-08

    Certain serovars of Salmonella enterica subsp. enterica cause invasive disease (e.g., enteric fever, bacteremia, septicemia, meningitis, etc.) in humans and constitute a global public health problem. A rapid, sensitive diagnostic test is needed to allow prompt initiation of therapy in individual patients and for measuring disease burden at the population level. An innovative and promising new rapid diagnostic technique is microwave-accelerated metal-enhanced fluorescence (MAMEF). We have adapted this assay platform to detect the chromosomal oriC locus common to all Salmonella enterica subsp. enterica serovars. We have shown efficient lysis of biologically relevant concentrations of Salmonella spp. suspended in bacteriological media using microwave-induced lysis. Following lysis and DNA release, as little as 1 CFU of Salmonella in 1 ml of medium can be detected in Salmonella serovars Typhi, Paratyphi A, Paratyphi B, Paratyphi C, Typhimurium, Enteritidis and Choleraesuis but does not detect Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae or Acinetobacter baumanii. We have also performed preliminary experiments using a synthetic Salmonella oriC oligonucleotide suspended in whole human blood and observed rapid detection when the sample was diluted 1:1 with PBS. These pre-clinical data encourage progress to the next step to detect Salmonella in blood (and other ordinarily sterile, clinically relevant body fluids).

  8. Global Gene-expression Analysis of the Response of Salmonella Enteritidis to Egg White Exposure Reveals Multiple Egg White-imposed Stress Responses

    Directory of Open Access Journals (Sweden)

    Florence Baron

    2017-05-01

    Full Text Available Chicken egg white protects the embryo from bacterial invaders by presenting an assortment of antagonistic activities that combine together to both kill and inhibit growth. The key features of the egg white anti-bacterial system are iron restriction, high pH, antibacterial peptides and proteins, and viscosity. Salmonella enterica serovar Enteritidis is the major pathogen responsible for egg-borne infection in humans, which is partly explained by its exceptional capacity for survival under the harsh conditions encountered within egg white. However, at temperatures up to 42°C, egg white exerts a much stronger bactericidal effect on S. Enteritidis than at lower temperatures, although the mechanism of egg white-induced killing is only partly understood. Here, for the first time, the impact of exposure of S. Enteritidis to egg white under bactericidal conditions (45°C is explored by global-expression analysis. A large-scale (18.7% of genome shift in transcription is revealed suggesting major changes in specific aspects of S. Enteritidis physiology: induction of egg white related stress-responses (envelope damage, exposure to heat and alkalinity, and translation shutdown; shift in energy metabolism from respiration to fermentation; and enhanced micronutrient provision (due to iron and biotin restriction. Little evidence of DNA damage or redox stress was obtained. Instead, data are consistent with envelope damage resulting in cell death by lysis. A surprise was the high degree of induction of hexonate/hexuronate utilization genes, despite no evidence indicating the presence of these substrates in egg white.

  9. Impacts of Salmonella enterica Serovar Typhimurium and Its speG Gene on the Transcriptomes of In Vitro M Cells and Caco-2 Cells

    Science.gov (United States)

    Wang, Ke-Chuan; Huang, Chih-Hung; Huang, Ching-Jou

    2016-01-01

    Microfold or membranous (M) cells are specialized intestinal epithelial cells responsible for host immunity. The speG mutant of Salmonella Typhimurium (S. Typhimurium) is a nonreplicating strain within human cells to be a candidate vaccine vector for interacting with M cells. We conducted this study to identify the genes are differently expressed between in vitro M cells and Caco-2 cells, and to determine whether S. Typhimurium and speG affect the transcriptomes of both cell types. In vitro M cells and Caco-2 cells were infected with wild-type (WT) S. Typhimurium, its ΔspeG mutant, or none for 1 h for RNA microarrays; the transcriptomes among the 6 pools were pairwisely compared. Genetic loci encoding scaffold (e.g., HSCHR7_CTG4_4, HSCHR9_CTG9_35), long noncoding RNA, membrane-associated protein (PITPNB), neuron-related proteins (OR8D1, OR10G9, and NTNG2), and transporter proteins (MICU2 and SLC28A1) were significantly upregulated in uninfected M cells compared with uninfected Caco-2 cells; and their encoding proteins are promising M-cell markers. Significantly upregulated HSCHR7_CTG4_4 of uninfected in vitro M cells were speG-independently downregulated by S. Typhimurium infection that is a remarkable change representing an important but unreported characteristic of M cells. The immune responses of in vitro M cells and Caco-2 cells can differ and reply on speG or not, with speG-dependent regulation of KYL4, SCTR, IL6, TNF, and CELF4 in Caco-2 cells, JUN, KLF6, and KCTD11 in M cells, or speG-independent modulation of ZFP36 in both cells. This study facilitates understanding of the immune responses of in vitro M cells after administering the S. Typhimurium ΔspeG mutant as a future vaccine vector. PMID:27064787

  10. Polymorphism of genes encoding PmrAB in colistin-resistant strains of Escherichia coli and Salmonella enterica isolated from poultry and swine.

    Science.gov (United States)

    Quesada, Alberto; Porrero, M Concepción; Téllez, Sonia; Palomo, Gonzalo; García, María; Domínguez, Lucas

    2015-01-01

    To detect the occurrence of low susceptibility to colistin (polymyxin E), a last-resort antimicrobial, among enterobacteria isolated from samples of animal origin (poultry and swine) and to find out the molecular basis of colistin resistance. Salmonella enterica and Escherichia coli were isolated from eggs and swine samples. Bacterial strains were screened for colistin resistance by using MIC determinations interpreted according to EUCAST recommendations. pmrAB genes were amplified by PCR from bacterial isolates and their sequences were characterized. Nine colistin-resistant strains were detected in a collection of 739 enterobacteria (S. enterica and E. coli) isolated from animal samples taken in different environments. Sequences encoding the PmrAB two-component sensor-regulator from two colistin-resistant E. coli strains isolated from swine faeces presented three non-synonymous polymorphisms, producing the variants 39S → I and 81R → S of PmrA and 161V → G of PmrB, among which the involvement of mutations in PmrA-81 and PmrB-161 in resistance to the antimicrobial had been previously shown. No variation at the protein level was detected after analysis of PmrAB sequences from seven colistin-resistant S. enterica strains. E. coli strains carrying mutations in PmrAB that confer resistance to polymyxins, which might have evolved in vivo and have been rarely detected, are described for the first time in enterobacteria isolated from animals. © The Author 2014. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  11. Global Gene-expression Analysis of the Response of Salmonella Enteritidis to Egg White Exposure Reveals Multiple Egg White-imposed Stress Responses.

    Science.gov (United States)

    Baron, Florence; Bonnassie, Sylvie; Alabdeh, Mariah; Cochet, Marie-Françoise; Nau, Françoise; Guérin-Dubiard, Catherine; Gautier, Michel; Andrews, Simon C; Jan, Sophie

    2017-01-01

    Chicken egg white protects the embryo from bacterial invaders by presenting an assortment of antagonistic activities that combine together to both kill and inhibit growth. The key features of the egg white anti-bacterial system are iron restriction, high pH, antibacterial peptides and proteins, and viscosity. Salmonella enterica serovar Enteritidis is the major pathogen responsible for egg-borne infection in humans, which is partly explained by its exceptional capacity for survival under the harsh conditions encountered within egg white. However, at temperatures up to 42°C, egg white exerts a much stronger bactericidal effect on S. Enteritidis than at lower temperatures, although the mechanism of egg white-induced killing is only partly understood. Here, for the first time, the impact of exposure of S. Enteritidis to egg white under bactericidal conditions (45°C) is explored by global-expression analysis. A large-scale (18.7% of genome) shift in transcription is revealed suggesting major changes in specific aspects of S. Enteritidis physiology: induction of egg white related stress-responses (envelope damage, exposure to heat and alkalinity, and translation shutdown); shift in energy metabolism from respiration to fermentation; and enhanced micronutrient provision (due to iron and biotin restriction). Little evidence of DNA damage or redox stress was obtained. Instead, data are consistent with envelope damage resulting in cell death by lysis. A surprise was the high degree of induction of hexonate/hexuronate utilization genes, despite no evidence indicating the presence of these substrates in egg white.

  12. Conservation of Salmonella infection mechanisms in plants and animals.

    Directory of Open Access Journals (Sweden)

    Adam Schikora

    Full Text Available Salmonella virulence in animals depends on effectors injected by Type III Secretion Systems (T3SSs. In this report we demonstrate that Salmonella mutants that are unable to deliver effectors are also compromised in infection of Arabidopsis thaliana plants. Transcriptome analysis revealed that in contrast to wild type bacteria, T3SS mutants of Salmonella are compromised in suppressing highly conserved Arabidopsis genes that play a prominent role during Salmonella infection of animals. We also found that Salmonella originating from infected plants are equally virulent for human cells and mice. These results indicate a high degree of conservation in the defense and infection mechanism of animal and plant hosts during Salmonella infection.

  13. Prevalence, antibiogram, molecular characterization and reduction trial of Salmonella typhimurium isolated from different fish species

    OpenAIRE

    Bayomi, Rasha M. El; Darwish, Wageh S.; El-Moaty, Amany M. Abd; Gad, Tamer M.

    2016-01-01

    Salmonella is a major cause of food-borne outbreaks and infections in many countries worldwide. The present work aimed to investigate the prevalence, antibiotic sensitivity and detection of virulence-associated genes in Salmonella typhimurium isolated from fish samples (tilapia, mullet and catfish) collected from Zagazig city markets, Egypt. Salmonella typhimurium was isolated only from tilapia with a percentage of 13.3%. Antibiotic susceptibility testing of Salmonella typhimurium revealed ma...

  14. Non-essential genes form the hubs of genome scale protein function and environmental gene expression networks in Salmonella enterica serovar Typhimurium

    DEFF Research Database (Denmark)

    Rosenkrantz, Jesper T.; Aarts, Henk; Abee, Tjakko

    2013-01-01

    in a high number of metabolic pathways and cell functions. The five genes with most connections in the transcriptional network (wraB, ygaU, uspA, cbpA and osmC) and in the genome scale network (ychN, siiF (STM4262), yajD, ybeB and dcoC) were selected for mutations, however mutagenesis of ygaU and ybe...... mutation but a greater susceptibility to directed attacks, however, we found that genes that formed hubs were dispensable for growth, stress adaptation and virulence, suggesting that evolution favors non-essential genes as main connectors in cellular networks....

  15. Identification of Novel Serodiagnostic Signatures of Typhoid Fever Using a Salmonella Proteome Array.

    Science.gov (United States)

    Darton, Thomas C; Baker, Stephen; Randall, Arlo; Dongol, Sabina; Karkey, Abhilasha; Voysey, Merryn; Carter, Michael J; Jones, Claire; Trappl, Krista; Pablo, Jozelyn; Hung, Chris; Teng, Andy; Shandling, Adam; Le, Tim; Walker, Cassidy; Molina, Douglas; Andrews, Jason; Arjyal, Amit; Basnyat, Buddha; Pollard, Andrew J; Blohmke, Christoph J

    2017-01-01

    Current diagnostic tests for typhoid fever, the disease caused by Salmonella Typhi, are poor. We aimed to identify serodiagnostic signatures of typhoid fever by assessing microarray signals to 4,445 S. Typhi antigens in sera from 41 participants challenged with oral S. Typhi. We found broad, heterogeneous antibody responses with increasing IgM/IgA signals at diagnosis. In down-selected 250-antigen arrays we validated responses in a second challenge cohort (n = 30), and selected diagnostic signatures using machine learning and multivariable modeling. In four models containing responses to antigens including flagellin, OmpA, HlyE, sipC, and LPS, multi-antigen signatures discriminated typhoid (n = 100) from other febrile bacteremia (n = 52) in Nepal. These models contained combinatorial IgM, IgA, and IgG responses to 5 antigens (ROC AUC, 0.67 and 0.71) or 3 antigens (0.87), although IgA responses to LPS also performed well (0.88). Using a novel systematic approach we have identified and validated optimal serological diagnostic signatures of typhoid fever.

  16. Identification of Novel Serodiagnostic Signatures of Typhoid Fever Using a Salmonella Proteome Array

    Directory of Open Access Journals (Sweden)

    Thomas C. Darton

    2017-09-01

    Full Text Available Current diagnostic tests for typhoid fever, the disease caused by Salmonella Typhi, are poor. We aimed to identify serodiagnostic signatures of typhoid fever by assessing microarray signals to 4,445 S. Typhi antigens in sera from 41 participants challenged with oral S. Typhi. We found broad, heterogeneous antibody responses with increasing IgM/IgA signals at diagnosis. In down-selected 250-antigen arrays we validated responses in a second challenge cohort (n = 30, and selected diagnostic signatures using machine learning and multivariable modeling. In four models containing responses to antigens including flagellin, OmpA, HlyE, sipC, and LPS, multi-antigen signatures discriminated typhoid (n = 100 from other febrile bacteremia (n = 52 in Nepal. These models contained combinatorial IgM, IgA, and IgG responses to 5 antigens (ROC AUC, 0.67 and 0.71 or 3 antigens (0.87, although IgA responses to LPS also performed well (0.88. Using a novel systematic approach we have identified and validated optimal serological diagnostic signatures of typhoid fever.

  17. Meta-analysis of Chicken – Salmonella infection experiments

    Directory of Open Access Journals (Sweden)

    te Pas Marinus FW

    2012-04-01

    Full Text Available Abstract Background Chicken meat and eggs can be a source of human zoonotic pathogens, especially Salmonella species. These food items contain a potential hazard for humans. Chickens lines differ in susceptibility for Salmonella and can harbor Salmonella pathogens without showing clinical signs of illness. Many investigations including genomic studies have examined the mechanisms how chickens react to infection. Apart from the innate immune response, many physiological mechanisms and pathways are reported to be involved in the chicken host response to Salmonella infection. The objective of this study was to perform a meta-analysis of diverse experiments to identify general and host specific mechanisms to the Salmonella challenge. Results Diverse chicken lines differing in susceptibility to Salmonella infection were challenged with different Salmonella serovars at several time points. Various tissues were sampled at different time points post-infection, and resulting host transcriptional differences investigated using different microarray platforms. The meta-analysis was performed with the R-package metaMA to create lists of differentially regulated genes. These gene lists showed many similarities for different chicken breeds and tissues, and also for different Salmonella serovars measured at different times post infection. Functional biological analysis of these differentially expressed gene lists revealed several common mechanisms for the chicken host response to Salmonella infection. The meta-analysis-specific genes (i.e. genes found differentially expressed only in the meta-analysis confirmed and expanded the biological functional mechanisms. Conclusions The meta-analysis combination of heterogeneous expression profiling data provided useful insights into the common metabolic pathways and functions of different chicken lines infected with different Salmonella serovars.

  18. Microarray-based detection of antibiotic resistance and virulence factor genes in Salmonella spp. isolated from food-producing animals and processed food

    OpenAIRE

    Figueiredo, Rui Filipe Ramos

    2016-01-01

    Tese de doutoramento em Ciências Farmacêuticas, na especialidade de Microbiologia e Parasitologia, apresentada à Faculdade de Farmácia da Universidade de Coimbra Salmonella enterica é uma bactéria patogénica de origem alimentar que infecta seres humanos pelo mundo inteiro. Nalguns casos, as infeções por Salmonella requerem tratamento com antibióticos. A resistência a agentes antimicrobianos é um problema global e leva ao insucesso do tratamento de infeções bacterianas. Alguns estudos têm s...

  19. 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. Published by Elsevier Ltd.

  20. Distribution of antibody titer against Salmonella enterica among healthy individuals in nepal

    Directory of Open Access Journals (Sweden)

    Koirala Janak

    2009-01-01

    Full Text Available Abstract Background Enteric fever is an endemic problem in Nepal and Widal agglutination test is widely used for its diagnosis but a normal baseline titer in healthy population and cutoff values have not been established. Methods We measured average baseline antibody titers against "O" and "H" antigens of Salmonella enterica serotype Typhi and "H" antigens of serotypes Paratyphi A and Paratyphi B among apparently healthy blood donors in Nepal. The antibody titers were measured using Standard Widal Confirmatory Quantitative Tube test. Results Among the 100 blood samples collected from healthy volunteers, 62 individuals had significant antibody titers (≥ 1:20 against one of the four antigens against S. enterica. Among 54 samples with an anti-O titer against serotype Typhi, 15 and 36 samples had titers of ≥ 1:60 and ≥ 1:40, respectively. A significant proportion (12% of all had anti-O titer of ≥ 1:80. Similarly, among the 59 samples demonstrating anti-H titers of ≥ 1:20 to S. enterica serotype Typhi, 29 had a titer of ≥ 1:80 and 12 had 1:160. For S. enterica serotypes Paratyphi A and B, anti-H titers of ≥ 1:20 were found only in 12% and 3%, respectively, of all samples tested. Conclusion When a single Widal agglutination titer is used for the diagnosis of enteric fever, it will be more appropriate to change the currently used cutoff levels against S. enterica serotype Typhi to > 1:80 for anti-O and > 1:160 for anti-H titers for Nepal.

  1. 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

    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

  2. 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

  3. Pathogenicity, Epidemiology and Virulence Factors of Salmonella species: A Review

    Directory of Open Access Journals (Sweden)

    Tamègnon Victorien DOUGNON

    2017-12-01

    Full Text Available Salmonella infections are major public health problems worldwide. The hereby review aimed to establish an overview on the pathogenicity, epidemiology and virulence factors of Salmonella spp. in the world. A systematic search was conducted online using the keywords ‘Salmonella’, ‘Salmonella spp.’, ‘Salmonella spp. Epidemiology’, ‘virulence factors of Salmonella spp. in the world’, ‘bacteria responsible for the contamination of meat products’, ‘non-typhoid salmonella’. These keywords were entered into databases such as PubMed and Google Scholar using mainly French language. The obtained articles were included based on the reliability of their source, the study area (usually Benin and Africa and the subject. The review revealed that Salmonella spp. is motile Gram-negative rod-shaped bacteria, of the family Enterobacteriaceae, currently counting more than 2,600 serovars. Human contamination occurs through the ingestion of contaminated water and food and can cause gastroenteritis or typhoid fever, which are two serious public health problems. A gene set constituting the pathogenicity islands determines the pathogenesis of Salmonella spp. The diagnosis is based on bacteriological, serological and molecular techniques. Salmonella infections are usually treated using antibiotics; however, emergence of antibiotic resistance in these microorganisms suggests that the anti-salmonella control should explore new sources such as medicinal plants

  4. [Epidemiological studies on salmonella in a particular area ("Walcheren Project"). III. The incidence of salmonella in man, insects, gulls as well as foods scrapings from butcher's blocks, effluents of sewage treatment plants and drains from butcher's shops (author's transl)].

    Science.gov (United States)

    Edel, W; Van Schothorst, M; Van Leusden, F M; Kampelmacher, E H

    1977-03-15

    In continuation of previous studies, various materials (meat and meat products, insects, gull droppings, scrapings from butcher's blocks, effluents of sewage treatment plants, drains from butcher's shops and faeces of patients) were examined again at the same time for the presence of Salmonella in a relatively small are (Walcheren) over a period of three months. As was also the case in previous studies, S. typhi murium (27.5 per cent), S. panama (22.2 per cent) and S. brandenburg (9.2 per cent) were the three serotypes most frequently isolated. The three most frequently isolated phage types of S. typhi murium were II 505 (62.1 per cent), II 502 (5.3 per cent) and I 650 (4.2 per cent). The serotypes and phage types were present in nearly all the materials studied which again emphasizes the fact that there are contamination cycles of Salmonella. These studies showed that the route of contamination divides in the butcher's shop. Salmonella ogranisms carried with the meat frome the slaughter-house find their way into the drains on the one hand, and, by meat and meat products, to consumers on the other. Moreover, the high degree of contamination of effluents is not in accordance with the small number of cases of salmonellosis.

  5. E. coli Nissle 1917 Affects Salmonella adhesion to porcine intestinal epithelial cells.

    Directory of Open Access Journals (Sweden)

    Peter Schierack

    Full Text Available BACKGROUND: The probiotic Escherichia coli strain Nissle 1917 (EcN has been shown to interfere in a human in vitro model with the invasion of several bacterial pathogens into epithelial cells, but the underlying molecular mechanisms are not known. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we investigated the inhibitory effects of EcN on Salmonella Typhimurium invasion of porcine intestinal epithelial cells, focusing on EcN effects on the various stages of Salmonella infection including intracellular and extracellular Salmonella growth rates, virulence gene regulation, and adhesion. We show that EcN affects the initial Salmonella invasion steps by modulating Salmonella virulence gene regulation and Salmonella SiiE-mediated adhesion, but not extra- and intracellular Salmonella growth. However, the inhibitory activity of EcN against Salmonella invasion always correlated with EcN adhesion capacities. EcN mutants defective in the expression of F1C fimbriae and flagellae were less adherent and less inhibitory toward Salmonella invasion. Another E. coli strain expressing F1C fimbriae was also adherent to IPEC-J2 cells, and was similarly inhibitory against Salmonella invasion like EcN. CONCLUSIONS: We propose that EcN affects Salmonella adhesion through secretory components. This mechanism appears to be common to many E. coli strains, with strong adherence being a prerequisite for an effective reduction of SiiE-mediated Salmonella adhesion.

  6. Construction of a bivalent DNA vaccine co-expressing S genes of transmissible gastroenteritis virus and porcine epidemic diarrhea virus delivered by attenuated Salmonella typhimurium.

    Science.gov (United States)

    Zhang, Yudi; Zhang, Xiaohui; Liao, Xiaodan; Huang, Xiaobo; Cao, Sanjie; Wen, Xintian; Wen, Yiping; Wu, Rui; Liu, Wumei

    2016-06-01

    Porcine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) can cause severe diarrhea in newborn piglets and led to significant economic losses. The S proteins are the main structural proteins of PEDV and TGEV capable of inducing neutralizing antibodies in vivo. In this study, a DNA vaccine SL7207 (pVAXD-PS1-TS) co-expressing S proteins of TGEV and PEDV delivered by attenuated Salmonella typhimurium was constructed and its immunogenicity in piglets was investigated. Twenty-day-old piglets were orally immunized with SL7207 (pVAXD-PS1-TS) at a dosage of 1.6 × 10(11) CFU per piglet and then booster immunized with 2.0 × 10(11) CFU after 2 weeks. Humoral immune responses, as reflected by virus neutralizing antibodies and specific IgG and sIgA, and cellular immune responses, as reflected by IFN-γ, IL-4, and lymphocyte proliferation, were evaluated. SL7207 (pVAXD-PS1-TS) simultaneously elicited immune responses against TGEV and PEDV after oral immunization. The immune levels started to increase at 2 weeks after immunization and increased to levels statistically significantly different than controls at 4 weeks post-immunization, peaking at 6 weeks and declined at 8 weeks. The humoral, mucosal, and cellular immune responses induced by SL7207 (pAXD-PS1-TS) were significantly higher than those of the PBS and SL7207 (pVAXD) (p < 0.01). In particular, the levels of IFN-γ and IL-4 were higher than those induced by the single-gene vaccine SL7207 (pVAXD-PS1) (p < 0.05). These results demonstrated that SL7207 (pVAXD-PS1-TS) possess the immunological functions of the two S proteins of TGEV and PEDV, indicating that SL7207 (pVAXD-PS1-TS) is a candidate oral vaccine for TGE and PED.

  7. Using current molecular techniques for rapid differentiation of ...

    African Journals Online (AJOL)

    Typhoid fever is responsible for the deaths of many people annually. However, conventional and timeconsuming detection methods for Salmonella Typhi still dominate. By using a molecular based approach, it was possible to identify Salmonella Typhi by amplifying two specific genes (viaB and tyv) and by using RFLP ...

  8. Prevalence of Nontyphoidal Salmonella and Salmonella Strains with Conjugative Antimicrobial-Resistant Serovars Contaminating Animal Feed in Texas.

    Science.gov (United States)

    Hsieh, Yi-Cheng; Poole, Toni L; Runyon, Mick; Hume, Michael; Herrman, Timothy J

    2016-02-01

    The objective of this study was to characterize 365 nontyphoidal Salmonella enterica isolates from animal feed. Among the 365 isolates, 78 serovars were identified. Twenty-four isolates (7.0%) were recovered from three of six medicated feed types. Three of these isolates derived from the medicated feed, Salmonella Newport, Salmonella Typhimurium var. O 5- (Copenhagen), and Salmonella Lexington var. 15+ (Manila), displayed antimicrobial resistance. Susceptibility testing revealed that only 3.0% (12) of the 365 isolates displayed resistance to any of the antimicrobial agents. These 12 isolates were recovered from unmedicated dry beef feed (n = 3), medicated dry beef feed (n = 3), cabbage culls (n = 2), animal protein products (n = 2), dry dairy cattle feed (n = 1), and fish meal (n = 1). Only Salmonella Newport and Salmonella Typhimurium var. O 5- (Copenhagen) were multidrug resistant. Both isolates possessed the IncA/C replicon and the blaCMY-2 gene associated with cephalosporin resistance. Plasmid replicons were amplified from 4 of 12 resistant isolates. Plasmids (40 kb) were Salmonella Montevideo and Salmonella Kentucky. Conjugation experiments were done using 7 of the 12 resistant isolates as donors. Only Salmonella Montevideo, possessing a plasmid and amplifying IncN, produced transconjugants. Transconjugants displayed the same antimicrobial resistance profile as did the donor isolate. Three isolates that amplified replicons corresponding to IncA/C or IncHI2 did not produce transconjugants at 30 or 37°C. The results of this study suggest that the prevalence of antimicrobial-resistant Salmonella contaminating animal feed is low in Texas. However, Salmonella was more prevalent in feed by-products; fish meal had the highest prevalence (84%) followed by animal protein products (48%). Ten of the 35 feed types had no Salmonella contamination. Further investigation is needed to understand the possible role of specific feed types in the dissemination of antimicrobial

  9. 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

    ciprofloxacin and gentamicin was low (n = 2; 0.9 %). Multiply resistant isolates included Salmonella Kentucky, the most resistant serovar. qnrB19 was found in two isolates of Salmonella Corvallis and one isolate of Salmonella Larochelle, respectively, while qnrS1 was found in two isolates of Salmonella Derby...... 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...... Kingston (n = 13; 5.7 %). The most widely distributed serovars among the farms were Salmonella Give (six farms) and Salmonella Elisaberthville (six farms). Resistance to chloramphenicol, sulfonamides, nalidixic acid, streptomycin, and tetracycline ranged from 11.6 % (n = 26) to 22.8 % (n = 51). Resistance...

  10. Attachment of Salmonella spp. to pork meat

    DEFF Research Database (Denmark)

    Hansen, Trine; Riber, Leise; Löfström, Charlotta

    2011-01-01

    Five strains of Salmonella, one wildtype and four knock-out mutants (the prg, flhDC, yhjH and fliC genes) were investigated based on their probability to attach and subsequently detach from a surface of pork fillet. The attachment followed by detachment was measured and modelled for two different...

  11. Adaptation of red blood cell lysis represents a fundamental breakthrough that improves the sensitivity of Salmonella detection in blood

    Science.gov (United States)

    Boyd, MA; Tennant, SM; Melendez, JH; Toema, D; Galen, JE; Geddes, CD; Levine, MM

    2015-01-01

    Aims Isolation of Salmonella Typhi from blood culture is the standard diagnostic for confirming typhoid fever but it is unavailable in many developing countries. We previously described a Microwave Accelerated Metal Enhanced Fluorescence (MAMEF)-based assay to detect Salmonella in medium. Attempts to detect Salmonella in blood were unsuccessful, presumably due to the interference of erythrocytes. The objective of this study was to evaluate various blood treatment methods that could be used prior to PCR, real-time PCR or MAMEF to increase sensitivity of detection of Salmonella. Methods and Results We tested ammonium chloride and erythrocyte lysis buffer, water, Lymphocyte Separation Medium, BD Vacutainer® CPT™ Tubes and dextran. Erythrocyte lysis buffer was the best isolation method as it is fast, inexpensive and works with either fresh or stored blood. The sensitivity of PCR- and real-time PCR detection of Salmonella in spiked blood was improved when whole blood was first lysed using erythrocyte lysis buffer prior to DNA extraction. Removal of erythrocytes and clotting factors also enabled reproducible lysis of Salmonella and fragmentation of DNA, which are necessary for MAMEF sensing. Conclusions Use of the erythrocyte lysis procedure prior to DNA extraction has enabled improved sensitivity of Salmonella detection by PCR and real-time PCR and has allowed lysis and fragmentation of Salmonella using microwave radiation (for future detection by MAMEF). Significance and Impact of the Study Adaptation of the blood lysis method represents a fundamental breakthrough that improves the sensitivity of DNA-based detection of Salmonella in blood. PMID:25630831

  12. Typhoid fever: “You can’t hit what you can’t see”

    OpenAIRE

    Wangdi, Tamding; Winter, Sebastian E.; Bäumler, Andreas J.

    2012-01-01

    The host restricts dissemination of invasive enteric pathogens, such as non-typhoidal Salmonella serovars, by mounting acute inflammatory responses characterized by the recruitment of neutrophils. However, some enteric pathogens, such as Salmonella enterica serovar Typhi (S. typhi), can bypass these defenses and cause an invasive bloodstream infection known as typhoid fever. Recent studies on virulence mechanisms of S. typhi suggest that tight regulation of virulence gene expression during th...

  13. O-Serotype Conversion in Salmonella Typhimurium Induces Protective Immune Responses against Invasive Non-Typhoidal Salmonella Infections.

    Science.gov (United States)

    Li, Pei; Liu, Qing; Luo, Hongyan; Liang, Kang; Yi, Jie; Luo, Ying; Hu, Yunlong; Han, Yue; Kong, Qingke

    2017-01-01

    Salmonella infections remain a big problem worldwide, causing enteric fever by Salmonella Typhi (or Paratyphi) or self-limiting gastroenteritis by non-typhoidal Salmonella (NTS) in healthy individuals. NTS may become invasive and cause septicemia in elderly or immuno-compromised individuals, leading to high mortality and morbidity. No vaccines are currently available for preventing NTS infection in human. As these invasive NTS are restricted to several O-antigen serogroups including B1, D1, C1, and C2, O-antigen polysaccharide is believed to be a good target for vaccine development. In this study, a strategy of O-serotype conversion was investigated to develop live attenuated S. Typhimurium vaccines against the major serovars of NTS infections. The immunodominant O4 serotype of S. Typhimurium was converted into O9, O7, and O8 serotypes through unmarked chromosomal deletion-insertion mutations. O-serotype conversion was confirmed by LPS silver staining and western blotting. All O-serotype conversion mutations were successfully introduced into the live attenuated S. Typhimurium vaccine S738 (Δcrp Δcya) to evaluate their immunogenicity in mice model. The vaccine candidates induced high amounts of heterologous O-polysaccharide-specific functional IgG responses. Vaccinated mice survived a challenge of 100 times the 50% lethality dose (LD50) of wild-type S. Typhimurium. Protective efficacy against heterologous virulent Salmonella challenges was highly O-serotype related. Furthermore, broad-spectrum protection against S. Typhimurium, S. Enteritidis, and S. Choleraesuis was observed by co-vaccination of O9 and O7 O-serotype-converted vaccine candidates. This study highlights the strategy of expressing heterologous O-polysaccharides via genetic engineering in developing live attenuated S. Typhimurium vaccines against NTS infections.

  14. O-Serotype Conversion in Salmonella Typhimurium Induces Protective Immune Responses against Invasive Non-Typhoidal Salmonella Infections

    Directory of Open Access Journals (Sweden)

    Pei Li

    2017-12-01

    Full Text Available Salmonella infections remain a big problem worldwide, causing enteric fever by Salmonella Typhi (or Paratyphi or self-limiting gastroenteritis by non-typhoidal Salmonella (NTS in healthy individuals. NTS may become invasive and cause septicemia in elderly or immuno-compromised individuals, leading to high mortality and morbidity. No vaccines are currently available for preventing NTS infection in human. As these invasive NTS are restricted to several O-antigen serogroups including B1, D1, C1, and C2, O-antigen polysaccharide is believed to be a good target for vaccine development. In this study, a strategy of O-serotype conversion was investigated to develop live attenuated S. Typhimurium vaccines against the major serovars of NTS infections. The immunodominant O4 serotype of S. Typhimurium was converted into O9, O7, and O8 serotypes through unmarked chromosomal deletion–insertion mutations. O-serotype conversion was confirmed by LPS silver staining and western blotting. All O-serotype conversion mutations were successfully introduced into the live attenuated S. Typhimurium vaccine S738 (Δcrp Δcya to evaluate their immunogenicity in mice model. The vaccine candidates induced high amounts of heterologous O-polysaccharide-specific functional IgG responses. Vaccinated mice survived a challenge of 100 times the 50% lethality dose (LD50 of wild-type S. Typhimurium. Protective efficacy against heterologous virulent Salmonella challenges was highly O-serotype related. Furthermore, broad-spectrum protection against S. Typhimurium, S. Enteritidis, and S. Choleraesuis was observed by co-vaccination of O9 and O7 O-serotype-converted vaccine candidates. This study highlights the strategy of expressing heterologous O-polysaccharides via genetic engineering in developing live attenuated S. Typhimurium vaccines against NTS infections.

  15. Isolation, amplification and characterization of foodborne pathogen disease bacteria gene for rapid kit test development

    Science.gov (United States)

    Nurjayadi, M.; Santoso, I.; Kartika, I. R.; Kurniadewi, F.; Saamia, V.; Sofihan, W.; Nurkhasanah, D.

    2017-07-01

    There is a lot of public concern over food safety. Food-safety cases recently, including many food poisoning cases in both the developed and developing countries, considered to be the national security threats which involved police investigation. Quick and accurate detection methods are needed to handle the food poisoning cases with a big number of sufferers at the same time. Therefore, the research is aimed to develop a specific, sensitive, and rapid result molecular detection tool for foodborne pathogen bacteria. We, thus, propose genomic level approach with Polymerase Chain Reaction. The research has successfully produced a specific primer to perform amplification to fim-C S. typhi, E. coli, and pef Salmonella typhimurium genes. The electrophoresis result shows that amplification products are 95 base pairs, 121 base pairs, and 139 base pairs; and all three genes are in accordance with the size of the in silico to third genes bacteria. In conclusion, the research has been successfully designed a specific detection tool to three foodborne pathogen bacteria genes. Further stages test and the uses of Real-time PCR in the detection are still in the trial process for better detection method.

  16. Genomics of Salmonella Species

    Science.gov (United States)

    Canals, Rocio; McClelland, Michael; Santiviago, Carlos A.; Andrews-Polymenis, Helene

    Progress in the study of Salmonella survival, colonization, and virulence has increased rapidly with the advent of complete genome sequencing and higher capacity assays for transcriptomic and proteomic analysis. Although many of these techniques have yet to be used to directly assay Salmonella growth on foods, these assays are currently in use to determine Salmonella factors necessary for growth in animal models including livestock animals and in in vitro conditions that mimic many different environments. As sequencing of the Salmonella genome and microarray analysis have revolutionized genomics and transcriptomics of salmonellae over the last decade, so are new high-throughput sequencing technologies currently accelerating the pace of our studies and allowing us to approach complex problems that were not previously experimentally tractable.

  17. Ultra-Fast and Sensitive Detection of Non-Typhoidal Salmonella Using Microwave-Accelerated Metal-Enhanced Fluorescence (“MAMEF”)

    Science.gov (United States)

    Galen, James E.; Geddes, Chris D.; Levine, Myron M.

    2011-01-01

    Certain serovars of Salmonella enterica subsp. enterica cause invasive disease (e.g., enteric fever, bacteremia, septicemia, meningitis, etc.) in humans and constitute a global public health problem. A rapid, sensitive diagnostic test is needed to allow prompt initiation of therapy in individual patients and for measuring disease burden at the population level. An innovative and promising new rapid diagnostic technique is microwave-accelerated metal-enhanced fluorescence (MAMEF). We have adapted this assay platform to detect the chromosomal oriC locus common to all Salmonella enterica subsp. enterica serovars. We have shown efficient lysis of biologically relevant concentrations of Salmonella spp. suspended in bacteriological media using microwave-induced lysis. Following lysis and DNA release, as little as 1 CFU of Salmonella in 1 ml of medium can be detected in Salmonella serovars Typhi, Paratyphi A, Paratyphi B, Paratyphi C, Typhimurium, Enteritidis and Choleraesuis but does not detect Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae or Acinetobacter baumanii. We have also performed preliminary experiments using a synthetic Salmonella oriC oligonucleotide suspended in whole human blood and observed rapid detection when the sample was diluted 1∶1 with PBS. These pre-clinical data encourage progress to the next step to detect Salmonella in blood (and other ordinarily sterile, clinically relevant body fluids). PMID:21494634

  18. Two Novel Salmonella Bivalent Vaccines Confer Dual Protection against Two Salmonella Serovars in Mice

    Science.gov (United States)

    Zhao, Xinxin; Dai, Qinlong; Jia, Renyong; Zhu, Dekang; Liu, Mafeng; Wang, Mingshu; Chen, Shun; Sun, Kunfeng; Yang, Qiao; Wu, Ying; Cheng, Anchun

    2017-01-01

    Non-typhoidal Salmonella includes thousands of serovars that are leading causes of foodborne diarrheal illness worldwide. In this study, we constructed three bivalent vaccines for preventing both Salmonella Typhimurium and Salmonella Newport infections by using the aspartate semialdehyde dehydrogenase (Asd)-based balanced-lethal vector-host system. The constructed Asd+ plasmid pCZ11 carrying a subset of the Salmonella Newport O-antigen gene cluster including the wzx-wbaR-wbaL-wbaQ-wzy-wbaW-wbaZ genes was introduced into three Salmonella Typhimurium mutants: SLT19 (Δasd) with a smooth LPS phenotype, SLT20 (Δasd ΔrfbN) with a rough LPS phenotype, and SLT22 (Δasd ΔrfbN ΔpagL::T araC PBAD rfbN) with a smooth LPS phenotype when grown with arabinose. Immunoblotting demonstrated that SLT19 harboring pCZ11 [termed SLT19 (pCZ11)] co-expressed the homologous and heterologous O-antigens; SLT20 (pCZ11) exclusively expressed the heterologous O-antigen; and when arabinose was available, SLT22 (pCZ11) expressed both types of O-antigens, while in the absence of arabinose, SLT22 (pCZ11) expressed only the heterologous O-antigen. Exclusive expression of the heterologous O-antigen in Salmonella Typhimurium decreased the swimming ability of the bacterium and its susceptibility to polymyxin B. Next, the crp gene was deleted from the three recombinant strains for attenuation purposes, generating the three bivalent vaccine strains SLT25 (pCZ11), SLT26 (pCZ11), and SLT27 (pCZ11), respectively. Groups of BALB/c mice (12 mice/group) were orally immunized with 109 CFU of each vaccine strain twice at an interval of 4 weeks. Compared with a mock immunization, immunization with all three vaccine strains induced significant serum IgG responses against both Salmonella Typhimurium and Salmonella Newport LPS. The bacterial loads in the mouse tissues were significantly lower in the three vaccine-strain-immunized groups than in the mock group after either Salmonella Typhimurium or Salmonella

  19. Two Novel Salmonella Bivalent Vaccines Confer Dual Protection against Two Salmonella Serovars in Mice

    Directory of Open Access Journals (Sweden)

    Xinxin Zhao

    2017-09-01

    Full Text Available Non-typhoidal Salmonella includes thousands of serovars that are leading causes of foodborne diarrheal illness worldwide. In this study, we constructed three bivalent vaccines for preventing both Salmonella Typhimurium and Salmonella Newport infections by using the aspartate semialdehyde dehydrogenase (Asd-based balanced-lethal vector-host system. The constructed Asd+ plasmid pCZ11 carrying a subset of the Salmonella Newport O-antigen gene cluster including the wzx-wbaR-wbaL-wbaQ-wzy-wbaW-wbaZ genes was introduced into three Salmonella Typhimurium mutants: SLT19 (Δasd with a smooth LPS phenotype, SLT20 (Δasd ΔrfbN with a rough LPS phenotype, and SLT22 (Δasd ΔrfbN ΔpagL::T araC PBADrfbN with a smooth LPS phenotype when grown with arabinose. Immunoblotting demonstrated that SLT19 harboring pCZ11 [termed SLT19 (pCZ11] co-expressed the homologous and heterologous O-antigens; SLT20 (pCZ11 exclusively expressed the heterologous O-antigen; and when arabinose was available, SLT22 (pCZ11 expressed both types of O-antigens, while in the absence of arabinose, SLT22 (pCZ11 expressed only the heterologous O-antigen. Exclusive expression of the heterologous O-antigen in Salmonella Typhimurium decreased the swimming ability of the bacterium and its susceptibility to polymyxin B. Next, the crp gene was deleted from the three recombinant strains for attenuation purposes, generating the three bivalent vaccine strains SLT25 (pCZ11, SLT26 (pCZ11, and SLT27 (pCZ11, respectively. Groups of BALB/c mice (12 mice/group were orally immunized with 109 CFU of each vaccine strain twice at an interval of 4 weeks. Compared with a mock immunization, immunization with all three vaccine strains induced significant serum IgG responses against both Salmonella Typhimurium and Salmonella Newport LPS. The bacterial loads in the mouse tissues were significantly lower in the three vaccine-strain-immunized groups than in the mock group after either Salmonella Typhimurium or

  20. Molecular characterization of "inconsistent" variants of Salmonella Typhimurium isolated in Italy

    DEFF Research Database (Denmark)

    Barco, Lisa; Longo, Alessandra; Lettini, Antonia Anna

    2014-01-01

    Salmonella 4,[5],12:i:- is a variant of Salmonella Typhimurium, which lacks the expression of phase-2 flagellar antigen, generally associated with the deletion of the fljB gene. Additional mechanisms involving the fljAB operon ( fljA, fljB, and hin genes) lead to the lack of expression of phase-2...

  1. Meningitis Caused by Salmonella Newport in a Five-Year-Old Child

    Directory of Open Access Journals (Sweden)

    Ana De Malet

    2016-01-01

    Full Text Available Salmonella Newport is a Gram-negative bacillus belonging to the Enterobacteria family and the nontyphi Salmonella (NTS, usually related to gastroenteritis. Main difference between NTS and Salmonella typhi is that the last one evolves to an invasive disease easier than NTS. These can progress to bacteremias in around 5% of cases and secondary focuses can appear occasionally, as in meningitis. An infection of the central nervous system is uncommon, considering its incidence in 0.6–8% of the cases; most of them are described in developing countries and mainly in childhood, especially neonates. Bacterial meningitis by NTS mostly affects immunosuppressed people in Europe. Prognosis is adverse, with a 50% mortality rate, mainly due to complications of infection: hydrocephalus, ventriculitis, abscesses, subdural empyema, or stroke. Choice antibiotic treatments are cefotaxime, ceftriaxone, or ceftazidime. The aim of this paper is to present a case of meningitis caused by Salmonella Newport diagnosed in a five-year-old girl living in a rural area of the province of Ourense (Spain, with favorable evolution and without neurological disorders.

  2. Laboratory surveillance of Salmonella enterica from human clinical cases in Colombia 2005-2011.

    Science.gov (United States)

    Rodríguez, Edna Catering; Díaz-Guevara, Paula; Moreno, Jaime; Bautista, Adriana; Montaño, Lucy; Realpe, María Elena; Della Gaspera, Anabella; Wiesner, Magdalena

    Salmonella is an enteropathogen acquired through contaminated food or water. In Colombia, Salmonella spp. is included in the national surveillance of Acute Diarrhoeal Diseases and typhoid fever initiated in 1997. This report shows the phenotype and genotype results obtained from 2005 to 2011. A total of 4010 isolates of Salmonella enterica were analysed by serotyping with Kauffmann-White-LeMinor, antimicrobial resistance patterns, and pulse-field gel electrophoresis (PFGE). A total of 93 serovars were identified, of which, Typhimurium, Enteritidis, Typhi, Dublin, Panama, Derby, Braenderup, Saintpaul, an