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Detection of InvA Gene in Isolated Salmonella from Broilers by PCR Method  

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Full Text Available The presence of Salmonella was detected in 192 samples of poultry carcasses from poultry farms in Shiraz province (Iran. A total of 30 Salmonella isolates were found in chicken samples (15.6%, by conventional culturing and confirmed by PCR and serology methods. Strains of serogroup D1 were the most prevalent strains, followed by serogroups C1, B and C2. All strains were subjected to Salmonella-specific gene (invA and were confirmed as Salmonella positive by the predicted product a 284-bp DNA fragment.

T. Zahraei Salehi

2005-01-01

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Presencia del gen de invasividad inv A en cepas de Salmonella spp: aisladas de alimentos del Caribe Colombiano / Presence of the invasive gene invA in Salmonella spp: strains isolated from food in several cities of the Colombian Caribbean area  

Scientific Electronic Library Online (English)

Full Text Available SciELO Cuba | Language: Spanish Abstract in spanish Objetivo: establecer la presencia del gen de invasividad invA en aislamientos de Salmonella spp. obtenidos de alimentos en la región Caribe Colombiana. Métodos: se realizó un estudio microbiológico de control de alimentos en 4 ciudades de la región Caribe entre enero de 2002 y marzo de 2003. Se anal [...] izaron 1 300 muestras de alimentos provenientes de mercados y ventas callejeras. Resultados: se recuperaron 74 aislamientos de Salmonella spp. , en carne de res 30 (40,5 %), embutidos 13 (17,6 %), pollo 12 (16,2 %), queso 9 (12,2 %), cerdo 6 (8,1 %) y otros 4 (5,5 %). Los serotipos más frecuentes fueron: S. anatum 14 (18,9 %), S. uganda 13 (17,6 %), S. newport 9 (12,2 %) y S. typhimurium 7 (9,5 %). El cebador invA amplificó un fragmento de 378 pb, el gen invA se detectó en 72 (97,3 %) aislamientos de Salmonella. Conclusiones: se detectó la presencia del gen invA en los serotipos de Salmonella circulantes en alimentos en la región Caribe Colombiana. Las implicaciones epidemiológicas de estos resultados permiten sugerir a las autoridades sanitarias tomar medidas estrictas en el control, prevención y diagnóstico de la infección por Salmonella en esta región Abstract in english Objective: to establish the presence of invasive gene invA in Salmonella spp. strains obtained from food in several cities of the Colombian Caribbean area. Methods: from January 2002 to March 2003, a microbiological study of quality control of food was carried out in four cities of the Colombian Car [...] ibbean area. One thousand and three hundred food samples were analyzed in fast food outlets located in city squares or markets. Results: seventy four isolates of Salmonella were recovered: 30 (40.5) in meat; 13 (17.6 %) in sausage; 12 (16.2 %) in chicken; 9 (12.2 %) in cheese; 6 (8.1 %) in pork and 4 (5.5 %) in other types of food. The most frequently isolated serotypes were S.anatum in 14 (18.9 %), S.uganda in 13 (17.6 %), S. newport in 9 (12.2 %) y S. typhimurium in 7 (9.5 %). The invA primer amplified 378 pb fragment, invA gene was detected in 72 (97.3 %) Salmonella isolates. Conclusions: it was possible to detect the invA gene in circulating serotypes of Salmonella isolates obtained from food in the Colombian Caribbean area, the epidemiological implications allow the health authorities to take measure for the prevention, control and diagnosis of Salmonella infection in the Colombian Caribbean area

Paula, Espinal Marin; Edgar, Prieto Suárez; Vanessa, Otero Jiménez; Salim, Máttar Velilla.

2006-06-01

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Salmonella enteritidis-specific monoclonal antibodies.  

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Three monoclonal antibodies (MAbs) were derived that are specific for Salmonella enteritidis. Such antibodies are of interest because reagents that specifically identify S. enteritidis are potentially useful for the diagnosis and detection of this pathogen. Immunization of BALB/c mice with intact, unfixed, ultraviolet-killed S. enteritidis permitted the derivation of a collection of hybridomas among which were found three MAbs: 1053, 1110, and 1170. Each MAb reacted with six independent field isolates of S. enteritidis, including phage type 4. However, none of these S. enteritidis-specific MAbs reacted with any of the following members of a broad diversity of Salmonella species: S. typhimurium, S. pullorum, S. berta, S. agona, S. dublin, S. miami, S. heidelberg, S. montevideo, S. senftenberg, and S. schwarzengrund. The S. enteritidis-specific determinant recognized by these MAbs is heat-labile, and preliminary experiments indicate that at least two of the MAbs recognize the same determinant. PMID:1378262

Lin, A W; Goldsby, R A; Snoeyenbos, G H

1992-01-01

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RAPID DETECTION OF Salmonella IN SHRIMP BY POLYMERASE CHAIN REACTION [Deteksi Cepat Salmonella pada Udang dengan Polymerase Chain Reaction  

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Full Text Available Shrimp is an important non-oil commodity for foreign trade in Indonesia. However, rejection of shrimp exports by the importing countries is still commonly encountered. In 2011, the USFDA recorded two cases of Salmonella spp. contamination in shrimp products from two shrimp processing companies in Indonesia. Analysis of Salmonella spp. in seafood is generally performed using a conventional method which takes at least 5 days. The objective of the study is to get a Salmonellae rapid detection method in shrimp by PCR. In this study, optimization of PCR protocol method to detect Salmonella invA gene was conducted using six different annealing temperatures (59, 59.5, 60.8, 62, 64 and 64.5°C. The results showed that 64°C was the optimum annealing temperature to detect the 284 bp fragment of Salmonella invA gene. The PCR based detection method has a DNA detection limit of 27.81ug/mL and 10°CFU/mL of viable salmonellae with 100% specificity. The PCR protocol is capable of detecting six different Salmonella serovars (S. Enteritidis, S. Hadar, S. Heidelberg, S. Kentucky, S. Paratyphi and S. Typhimurium but none of the non salmonellae isolates. Application of the PCR assay on Salmonella in shrimp after the selective enrichment step suggested that all 16 samples were positive for Salmonella. At the same time, the conventional method could only detected 3 samples for Salmonella positive.

Ulfah Amalia

2014-06-01

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InvA Protein Is a Nudix Hydrolase Required for Infection by Pathogenic Leptospira in Cell Lines and Animals*  

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Leptospirosis caused by pathogenic species of the genus Leptospira is a re-emerging zoonotic disease, which affects a wide variety of host species and is transmitted by contaminated water. The genomes of several pathogenic Leptospira species contain a gene named invA, which contains a Nudix domain. However, the function of this gene has never been characterized. Here, we demonstrated that the invA gene was highly conserved in protein sequence and present in all tested pathogenic Leptospira species. The recombinant InvA protein of pathogenic L. interrogans strain Lai hydrolyzed several specific dinucleoside oligophosphate substrates, reflecting the enzymatic activity of Nudix in Leptospira species. Pathogenic leptospires did not express this protein in media but temporarily expressed it at early stages (within 60 min) of infection of macrophages and nephric epithelial cells. Comparing with the wild type, the invA-deficient mutant displayed much lower infectivity and a significantly reduced survival rate in macrophages and nephric epithelial cells. Moreover, the invA-deficient leptospires presented an attenuated virulence in hamsters, caused mild histopathological damage, and were transmitted in lower numbers in the urine, compared with the wild-type strain. The invA revertant, made by complementing the invA-deficient mutant with the invA gene, reacquired virulence similar to the wild type in vitro and in vivo. The LD50 in hamsters was 1000-fold higher for the invA-deficient mutant than for the invA revertant and wild type. These results demonstrate that the InvA protein is a Nudix hydrolase, and the invA gene is essential for virulence in pathogenic Leptospira species. PMID:21862592

Luo, Yihui; Liu, Yan; Sun, Dexter; Ojcius, David M.; Zhao, Jinfang; Lin, Xuai; Wu, Dong; Zhang, Rongguang; Chen, Ming; Li, Lanjuan; Yan, Jie

2011-01-01

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Evaluation of different analysis and identification methods for Salmonella detection in surface drinking water sources  

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The standard method for detecting Salmonella generally analyzes food or fecal samples. Salmonella often occur in relatively low concentrations in environmental waters. Therefore, some form of concentration and proliferation may be needed. This study compares three Salmonella analysis methods and develops a new Salmonella detection procedure for use in environmental water samples. The new procedure for Salmonella detection include water concentration, nutrient broth enrichment, selection of Salmonella containing broth by PCR, isolation of Salmonella strains by selective culture plates, detection of possible Salmonella isolate by PCR, and biochemical testing. Serological assay and pulsed-field gel electrophoresis (PFGE) can be used to identify Salmonella serotype and genotype, respectively. This study analyzed 116 raw water samples taken from 18 water plants and belonging to 5 watersheds. Of these 116, 10 water samples (8.6%) taken from 7 water plants and belonging to 4 watersheds were positive for a Salmonella-specific polymerase chain reaction targeting the invA gene. Guided by serological assay results, this study identified 7 cultured Salmonella isolates as Salmonella enterica serovar: Alnaby, Enteritidis, Houten, Montevideo, Newport, Paratyphi B var. Java, and Victoria. These seven Salmonella serovars were identified in clinical cases for the same geographical areas, but only one of them was 100% homologous with clinical cases in the PFGE pattern. - Research highlights: ? A new Salmonella detecting procedure for environmental water is developed. ? Salmonella isolates are identified by serological assay and PFGE. ? A total of seven Salmonella serovars is isolated from environmental water.

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Rapid identification of Salmonella enterica serovars, Typhimurium, Choleraesuis, Infantis, Hadar, Enteritidis, Dublin and Gallinarum, by multiplex PCR.  

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Salmonella enterica subsp. enterica poses a threat to both human and animal health, with more than 2500 serovars having been reported to date. Salmonella serovars are identified by slide and tube agglutination tests using O and H antigen-specific anti-sera, although this procedure is both labor intensive and time consuming. Establishment of a method for rapid screening of the major Salmonella serovars is therefore required. We have established multiplex polymerase chain reaction (m-PCR) assays for identification of seven serovars of Salmonella, i.e., Typhimurium, Choleraesuis, Infantis, Hadar, Enteritidis, Dublin and Gallinarum. Three serovar-specific genomic regions (SSGRs) of each serovar were selected using an approach in comparative genomics. The Salmonella-specific invA gene was used to confirm the genetic background of the organisms. The isolates tested were identified as a target serovar when the three selected SSGRs and invA were all positive for amplification. The specificity of each m-PCR assay was investigated using 118 serovars of Salmonella and 12 species of non-Salmonella strains. Although a small number of false-positive results were observed in the m-PCR assays used to identify Typhimurium, Choleraesuis, Enteritidis and Dublin for closely related serovars, false-negative results were not observed in any assays. These assays had sufficient specificity to identify the seven Salmonella serovars, and therefore, have the potential for use as rapid screening methods. PMID:21329737

Akiba, Masato; Kusumoto, Masahiro; Iwata, Taketoshi

2011-04-01

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Differentially Evolved Genes of Salmonella Pathogenicity Islands: Insights into the Mechanism of Host Specificity in Salmonella  

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he species Salmonella enterica (S. enterica) includes many serovars that cause disease in avian and mammalian hosts. These serovars differ greatly in their host range and their degree of host adaptation. The host specificity of S. enterica serovars appears to be a complex phenomenon governed by multiple factors acting at different stages of the infection process, which makes identification of the cause/s of host specificity solely by experimental methods difficult.

Eswarappa, Sandeepa M.; Janice, Jessin; Nagarajan, Arvindhan G.; Balasundaram, Sudhagar V.; Karnam, Guruswamy; Dixit, Narendra M.; Chakravortty, Dipshikha

2008-01-01

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Differentially Evolved Genes of Salmonella Pathogenicity Islands: Insights into the Mechanism of Host Specificity in Salmonella  

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Background The species Salmonella enterica (S. enterica) includes many serovars that cause disease in avian and mammalian hosts. These serovars differ greatly in their host range and their degree of host adaptation. The host specificity of S. enterica serovars appears to be a complex phenomenon governed by multiple factors acting at different stages of the infection process, which makes identification of the cause/s of host specificity solely by experimental methods difficult. Methodology/Principal Findings In this study, we have employed a molecular evolution and phylogenetics based approach to identify genes that might play important roles in conferring host specificity to different serovars of S. enterica. These genes are ‘differentially evolved’ in different S. enterica serovars. This list of ‘differentially evolved’ genes includes genes that encode translocon proteins (SipD, SseC and SseD) of both Salmonella pathogenicity islands 1 and 2 encoded type three secretion systems, sptP, which encodes an effector protein that inhibits the mitogen-activated protein kinase pathway of the host cell, and genes which encode effector proteins (SseF and SifA) that are important in placing the Salmonella-containing vacuole in a juxtanuclear position. Conclusions/Significance Analysis of known functions of these ‘differentially evolved genes’ indicates that the products of these genes directly interact with the host cell and manipulate its functions and thereby confer host specificity, at least in part, to different serovars of S. enterica that are considered in this study. PMID:19050757

Eswarappa, Sandeepa M.; Janice, Jessin; Nagarajan, Arvindhan G.; Balasundaram, Sudhagar V.; Karnam, Guruswamy; Dixit, Narendra M.; Chakravortty, Dipshikha

2008-01-01

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LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP FOR THE DETECTION OF SALMONELLA SPP. ISOLATED FROM DIFFERENT FOOD TYPES  

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Full Text Available The objective of this study was the application and evaluation of a loop-mediated isothermal amplification (LAMP method for the detection of Salmonella spp. strains isolated from food samples. Salmonella specific invA gene sequences (50 strains, 15 serotypes were amplified at 65oC in 60 min. All of the strains of Salmonella subsp. Enterica were shown to be positive using the LAMP reaction assay, whereas, all other bacteria, virus and yeasts tested in this study were negative. LAMP products could be visually detected under day light or ultraviolet light, while the specific amplification of the DNA of Salmonella strains generated ladder-like pattern bands on agarose gel. LAMP is suitable for the sensitive, rapid, and inexpensive detection of Salmonella spp. in food analytical laboratories.

Kostas Papanotas

2012-08-01

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Comparison of commercial RNA extraction kits for preparation of DNA-free total RNA from Salmonella cells  

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Full Text Available Abstract Background The isolation of DNA-free RNA is a crucial step in the reverse transcription PCR (RT-PCR. Every RNA extraction procedure results in RNA samples contaminated with genomic DNA, which can cause false-positive outcomes in highly sensitive applications, including a recently developed quantitative real-time PCR (RT-qPCR assay that targets invA mRNA for the detection of live Salmonella cells. The assay of this specific mRNA can be used to indicate the presence of live, as opposed to dead, cells of Salmonella enterica in a food matrix. Findings We evaluated the ability of five RNA extraction kits to produce RNA preparations from exponentially growing Salmonella cells. The acceptability of the preparations for use in downstream applications such as RT-qPCR was judged in terms of the total amount of RNA recovered, the integrity of the RNA molecules, and minimal content of DNA. The five kits produced RNA preparations that differed markedly in yield, integrity of the Salmonella RNA and the amount of contaminant DNA. The greatest RNA recovery was achieved with the MasterPure kit; however, the preparation contained high levels of genomic DNA. The UltraClean extraction kit gave a low level of RNA recovery with a poor level of integrity. The RNeasy Mini, RiboPure and PureLink extraction kits produced high-quality, DNA-free RNA suitable for Salmonella detection by RT-qPCR. Conclusions We showed that the RNeasy Mini and PureLink RNA extraction kits were the most suitable for the detection of Salmonella invA mRNA by RT-qPCR. The use of these two kits will greatly reduce the frequency of false-positive results and might allow fast RT-qPCR determination of invA mRNA produced by viable Salmonella in food samples.

Gonzalez-Escalona Narjol

2010-07-01

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Multicenter validation of the analytical accuracy of Salmonella PCR: towards an international standard  

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As part of a major international project for the validation and standardization of PCR for detection of five major food-borne pathogens, four primer sets specific for Salmonella species were evaluated in-house for their analytical accuracy (selectivity and detection limit) in identifying 43 Salmonella spp. and 47 non-Salmonella strains. The most selective primer set was found to be 139-141 (K. Rahn, S. A. De Grandis, R. C. Clarke, S. A. McEwen, J. E. Galan, C. Ginocchio, R. Curtiss 111, and C. L. Gyles, Mol. Cell. Probes 6:271-279, 1992), which targets the invA gene. An extended determination of selectivity by using 364 strains showed that the inclusivity was 99.6% and the exclusivity was 100% for the invA primer set. To indicate possible PCR inhibitors derived from the sample DNA, an internal amplification control (IAC), which was coamplified with the invA target gene, was constructed. In the presence of 300 DNA copies of the IAC, the detection probability for primer set 139-141 was found to be 100% when a cell suspension containing 10(4) CFU/ml was used as the template in the PCR (50 CFU per reaction). The primer set was further validated in an international collaborative study that included 16 participating laboratories. Analysis with 28 coded ("blind") DNA samples revealed an analytical accuracy of 98%. Thus, a simple PCR assay that is specific for Salmonella spp. and amplifies a chromosomal DNA fragment detected by gel electrophoresis was established through extensive validation and is proposed as an international standard. This study addresses the increasing demand of quality assurance laboratories for standard diagnostic methods and presents findings that can facilitate the international comparison and exchange of epidemiological data.

Hoorfar, Jeffrey

2003-01-01

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Inhibition of the metabolism of streptococci and salmonella by specific antisera.  

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Streptococcal and salmonella antisera inhibited carbohydrate metabolism for groups A, B, C, and D streptococci and group E salmonella, as measured by the formation of [(14)C]dioxide from [(14)C]glucose metabolism. For salmonella, the inhibition was type specific since group E salmonella were inhibited only by salmonella E antisera and not by anti-salmonella A or C(1). For streptococci, quantitative differences were demonstrated, but major cross-reactivity was observed. At high concentrations, the antisera were bactericidal; at more dilute concentrations, for both salmonella and streptococci, carbohydrate metabolism was suppressed, but subculture on chocolate agar showed abundant growth. Cross-reacting antibodies could be absorbed by incubation with either antigen, e.g., streptococcal antisera versus heat-killed salmonella. The results suggest that the radiometric technique can be more sensitive than either capillary flocculation or visual detection of bacterial growth for detecting the inhibition of streptococci and salmonella by specific antibodies. The use of specific antisera may prove useful for bacterial species identification in an automated system for detection of bacterial growth. PMID:4823421

Larson, S M; Charache, P; Chen, M; Wagner, H N

1974-02-01

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Schistosoma-associated Salmonella resist antibiotics via specific fimbrial attachments to the flatworm  

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Full Text Available Abstract Background Schistosomes are parasitic helminths that infect humans through dermo-invasion while in contaminated water. Salmonella are also a common water-borne human pathogen that infects the gastrointestinal tract via the oral route. Both pathogens eventually enter the systemic circulation as part of their respective disease processes. Concurrent Schistosoma-Salmonella infections are common and are complicated by the bacteria adhering to adult schistosomes present in the mesenteric vasculature. This interaction provides a refuge in which the bacterium can putatively evade antibiotic therapy and anthelmintic monotherapy can lead to a massive release of occult Salmonella. Results Using a novel antibiotic protection assay, our results reveal that Schistosoma-associated Salmonella are refractory to eight different antibiotics commonly used to treat salmonellosis. The efficacy of these antibiotics was decreased by a factor of 4 to 16 due to this association. Salmonella binding to schistosomes occurs via a specific fimbrial protein (FimH present on the surface on the bacterium. This same fimbrial protein confers the ability of Salmonella to bind to mammalian cells. Conclusions Salmonella can evade certain antibiotics by binding to Schistosoma. As a result, effective bactericidal concentrations of antibiotics are unfortunately above the achievable therapeutic levels of the drugs in co-infected individuals. Salmonella-Schistosoma binding is analogous to the adherence of Salmonella to cells lining the mammalian intestine. Perturbing this binding is the key to eliminating Salmonella that complicate schistosomiasis.

Day Tim A

2011-06-01

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Detection of OmpA gene by PCR for specific detection of Salmonella serovars  

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Full Text Available Aim: The study was carried out to determine the sensitivity and specificity of OmpA gene in Salmonella serovarsthrough PCR.Materials and Methods: Aset of primers were designed targeting the OmpAgene specific for the Salmonella and polymerasechain reaction was standardized using Salomonella Typhimurium as a positive control and as a negative control 4 nonsalmonella cultures such as Campylobacter coli, Arcobacter butzleri, Brucella abortus and E. coli. Sensitivity of the test wasdetermined by serial dilution of genomic DNAof standard S. Typhimurium. The PCR standardized was used for screening 68strains of different serovars of Salmonella.Results: The PCR developed targeting OmpA specific for Salmonella was highly specific in detection of the salmonellaserovar alone and sensitivity was upto 68.8 fg. Atotal of 68 virulent/ natural strains of different serovars of salmonella takenup for the study were positive by OmpAbased PCR.Conclusions: This study reports that, OmpAgene which is conserved among Salmonella serovars can be used for the detectionof Salmonella in food or clinical samples in further studies, with high sensitivity and specificity.

Joy. L. Kataria

2013-10-01

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Comparison of PCR-ELISA and LightCycler real-time PCR assays for detecting Salmonella spp. in milk and meat samples  

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In a previous study, we reported the performance of a PCR assay amplifying 285-bp of the invA gene of Salmonella spp. through an international ring-trial involving four participating laboratories [Int. J. Food Microbiol. 89 (2003) 241]. Based on the validated set of primers and recent advancements in PCR technology, we have designed two specific PCR assays for detecting Salmonella spp. We have compared PCR-enzyme-linked immunosorbent assay (PCR-ELISA) and LightCycler real-time PCR assay (LC-PCR) with the standard ISO 6579 bacteriological reference method. The two PCR tests incorporated an internal amplification control (IAC) co-amplified with the invA gene of Salmonella to monitor potential PCR inhibitors and ensure successful amplification. The selectivity study involved 84 Salmonella and 44 non-Salmonella strains and the samples tested were represented by 60 artificially-contaminated samples of fish, minced beef and raw milk, and 92 naturally-contaminated milk and meat samples. When using either PCR-ELISA or LC-PCR assays, only Salmonella strains were detected. PCR-ELISA and LC-PCR assays gave with pure Salmonella cultures the same detection limit level of 10(3) CFU/ml, which corresponds respectively to 50 and 10 cells per PCR tube. Data on artificially contaminated samples indicated that both PCR methods were able to detect after enrichment less than five Salmonella cells in 25 g of food, giving 100% concordance with the ISO 6579 reference method. The results on naturally contaminated samples demonstrated that despite certain inhibition problems, LC-PCR and PCR-ELISA assays were highly specific and sensitive, and provide a powerful tool for detection of Salmonella in food samples.

Hoorfar, Jeffrey

2004-01-01

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Animal salmonelloses: a brief review of “host adaptation and host specificity” of Salmonella spp.  

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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

Grammato Evangelopoulou

2013-07-01

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PCR identification of Salmonella serogroups based on specific targets obtained by comparative genomics.  

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Comparative genomic approaches provide abundant information to reveal the diversity among Salmonella serogroups. In a local genomic sequence database, twenty-five Salmonella whole genomic sequences were divided into 6 (A, B, C1, C2, D and others) serogroups for mining the DNA fragments specific for serogroups A through D. For each serogroup, a reference sequence was selected and split into 1000-bp fragments in silico to align against all the other genomic sequences to obtain one or more serogroup-specific fragments. As a result, 2, 6, 7, 10, and 7 specific fragments were found for A, B, C1, C2 and D serogroups, respectively. Specific primer sets targeting these DNA fragments were designed for multiplex PCR assays identifying 21 Salmonella standard strains and 86 additional food isolates. The PCR results demonstrated good agreement with those from Salmonella serotyping. This means that the PCR assay may be able to identify 5 (A, B, C1, C2 and D) Salmonella serogroups and elucidate differences among them. Based on the gene annotations, the 32 serogroup-specific fragments were divided into 3 categories (membrane protein genes, rfb gene clusters, and fimbrial genes). Each gene from these three groups was conserved within the serogroup and was closely correlated with phenotypic characterization. This finding implies that these genes, which are associated with sugar synthesis and metabolism or glycosyl and O-actetyl transfer, impart the differences among Salmonella serogroups. PMID:21131088

Liu, Bin; Zhang, Lida; Zhu, Xinna; Shi, Chunlei; Chen, Jing; Liu, Weibing; He, Xiaohua; Shi, Xianming

2011-01-01

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Detection of Salmonella spp. using microsphere-based, fiber-optic DNA microarrays.  

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Salmonella spp. are one of the most problematic food pathogens in public health, as they are responsible for food poisoning associated with contamination of meat, poultry, and eggs. Thus, rapid and sensitive detection of Salmonella spp. is required to ensure food safety. In this study, a fiber-optic DNA microarray using microsphere-immobilized oligonucleotide probes specific for the Salmonella invA and spvB genes was developed for detection of Salmonella spp. Microarrays were prepared by randomly distributing DNA probe-functionalized microspheres (3.1-microm diameter) into microwells created by etching optical fiber bundles. Hybridization of the probe-functionalized microspheres to target DNA from Salmonella was performed and visualized using Cy3-labeled secondary probes in a sandwich-type assay format. In this study, 10(3)-10(4) cfu/mL of the target organism could be detected after 1-h hybridization without any additional amplification. The DNA microarray showed no cross-reactivity with other common food pathogens, including E. coli and Y. enterocolitica, and could even detect Salmonella spp. from cocktails of bacterial strains with only moderate loss of sensitivity due to nonspecific binding. This work suggests that fiber-optic DNA microarrays can be used for rapid and sensitive detection of Salmonella spp. Since fiber-optic microarrays can be prepared with different probes, this approach could also enable the simultaneous detection of multiple food pathogens. PMID:16053320

Ahn, Soohyoun; Walt, David R

2005-08-01

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Characterization and specificity of probiotics to prevent salmonella infection in mice  

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Full Text Available Background: Probiotic strains of bacteria can prevent Salmonella from causing disease by preventing the pathogen from colonizing the intestines. Two strains of probiotics, Lactobacillus acidophilius and Pediococcus spp, that were obtained from poultry fecal samples have been shown to be efficacious in poultry. The objective of this study was to determine if these strains of probiotics could prevent salmonellosis in a mouse model. Methods: First, both strains of probiotics were evaluated for in vitro efficacy to inhibit the growth of and interfere with virulence gene regulation in Salmonella enterica. For in vivo efficacy, mice was used which models Typhoid illness. Mice were divided into 2 groups: Control and treatment, Lactobacillus and Pediococcus (LP; 108 Log CFU. Two experiments were conducted. In the first experiment, the mice were treated with LP in water for the first two days of the experiment and challenged with Salmonella at day three. In the second experiment, the LP treatment was given in the water for 10 days and challenge was performed on day 11. In both experiments, at day 20 post-challenge, all mice were sacrificed, intestinal tracts and organs removed and cultured for Salmonella. Results: The probiotic strains inhibited the growth of Salmonella and down-regulation of virulence genes was noted, but dependent on the strain of Salmonella being evaluated. For the in vivo experiment, the probiotics did not afford the mice protection from infection and increasing the length of time the probiotics were administered did not improve the efficacy of the probiotics. Conclusions: It appears that these strains of probiotic bacteria are effective against Salmonella in vitro. However, these isolates did not afford protection from Salmonella infection to mice which may be due to host specifity as these isolates were obtained from poultry

Ana Andino

2014-08-01

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

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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.

Sun Qi

2009-11-01

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

Science.gov (United States)

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

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

2013-12-01

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A Conserved Domain in Type III Secretion Links the Cytoplasmic Domain of InvA to Elements of the Basal Body  

Energy Technology Data Exchange (ETDEWEB)

Protein type III secretion systems (T3SSs) are organic nanosyringes that achieve an energy-dependent translocation of bacterial proteins through the two membranes of Gram-negative organisms. Examples include the pathogenic systems of animals, plants and symbiotic bacteria that inject factors into eukaryotic cells, and the flagellar export system that secretes flagellin. T3SSs possess a core of several membrane-associated proteins that are conserved across all known bacterial species that use this system. The Salmonella protein InvA is one of the most highly conserved proteins of this core of critical T3SS components. The crystal structure of a C-terminal domain of InvA reveals an unexpected homology to domains that have been repeatedly found as building blocks of other elements of the T3SS apparatus. This suggests the surprising hypothesis that evolution has produced a significant component of the apparatus structure through a series of gene-duplication and gene-rearrangement events.

Lilic, M.; Quezada, C; Stebbins, C

2010-01-01

24

Eight-hour PCR-based procedure for the detection of Salmonella in raw oysters.  

Science.gov (United States)

The aim of this study was to evaluate the ability of a nested PCR system to detect Salmonella senftenberg in raw oysters. The specific primers of the PCR were derived from the invA gene sequence, essential for Salmonella invasiveness into epithelial cells. First, for the extraction of DNA, four methods (guanidine isothiocyanate, E.Z.N.A. Mollusc Kit, Chelex-100, and lysis with detergents) were compared. A nested PCR method combined with 3.5 h pre-enrichment in buffered peptone water (BPW) and DNA extraction by the resin Chelex-100 is proposed for the detection of S. senftenberg in oyster samples. The detection limit of the method is less than 0.1 CFU/ml (<1 CFU/g of oyster). This procedure is shown to be an excellent tool for the sensitive detection of S. senftenberg from naturally contaminated oysters, with results being obtained within 8 h. PMID:15668030

Vázquez-Novelle, M Dolores; Pazos, Antonio J; Abad, Marcelina; Sánchez, José L; Pérez-Parallé, M Luz

2005-02-01

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

Scientific Electronic Library Online (English)

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

M., Radhika; Majumder, Saugata; H.S., Murali; H.V., Batra.

26

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

Scientific Electronic Library Online (English)

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

M., Radhika; Majumder, Saugata; H.S., Murali; H.V., Batra.

2014-06-01

27

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

Science.gov (United States)

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

Radhika, M.; Saugata, Majumder; Murali, H.S.; Batra, H.V.

2014-01-01

28

Primers specific for the fimbrial major subunit gene stdA can be used to detect Salmonella enterica serovars.  

Science.gov (United States)

The feasibility of using two primers internal to the stdA gene (which encodes the fimbrial major subunit of the std fimbrial gene cluster in Salmonella enterica serovar Typhi) to detect Salmonella by PCR was explored. The 518-bp stdA specific sequence was conserved among 268 strains from 45 serovars of S. enterica. One Salmonella bongori CCUG 30042 strain and 34 non-Salmonella strains did not possess this sequence. A sensitivity test revealed that the stdA-specific primer set detected 3.4 x 10(-1) pg of genomic DNA and 3.0 x 10(5) CFU/ml with serial dilutions of Salmonella Typhimurium cells. In vitro testing for specificity using pig carcass sponge samples contaminated with Salmonella Typhimurium also was performed. An initial Salmonella Typhimurium inoculum of 4.4 x 10(1) CFU/ml in pig carcass exudates reached the stdA primer detection level after preenrichment in buffered peptone water at 37 degrees C for 18 h in the presence of indigenous non-Salmonella flora at 4.0 X 10(7) CFU/ml, but the detection level decreased to 4.4 x 10(0) CFU/ml after selective enrichment in Rappaport-Vassiliadis R10 broth for 18 h at 42 degrees C. The PCR method with primers specific for stdA is a quick and sensitive tool for detecting S. enterica, which is an important cause of foodborne disease. PMID:18592734

Chuang, Yin-Ching; Yang, Chia-Huei; Lin, Jiunn-Horng; Wang, Ke-Chuan; Cheng, Chun-Ping; Yeh, Kuang-Sheng

2008-06-01

29

Molecular epidemiology and in vitro antimicrobial susceptibility of Salmonella isolated from poultry in Kashmir.  

Science.gov (United States)

A total of 480 samples, comprising 429 faecal samples from healthy adult birds and 51 tissue samples from dead birds, were collected from four government poultry farms in the Kashmir valley from September 2007 to April 2008. In all, 33 Salmonella isolates were obtained. Of these, 28 (84.85%) isolates were Salmonella Gallinarum, 3 (9.09%) were Salmonella Enteritidis and the remaining 2 (6.06%) were Salmonella Typhimurium. All the isolates harboured the invA, sefA, stn and spvC virulence-specific genes. However, the sopB gene was found in only 90.9% of the isolates. Pulsed-field gel electrophoresis analysis of representative isolates revealed that the majority were related but a few belonged to different clones. The majority of the isolates were resistant to cefpodoxime, nalidixic acid and sulphadiazine and sensitive to chloramphenicol, cefotaxime and tetracycline. Isolation of multidrug-resistant Salmonella, including the zoonotically important serovars, revealed a potential threat not only to poultry but also to human health in Kashmir. PMID:21309466

Mir, I A; Wani, S A; Hussain, I; Qureshi, S D; Bhat, M A; Nishikawa, Y

2010-12-01

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Refining the LPS-Antigen in Salmonella Antibody Elisa for Poultry Enhanced Specificity without Impairing Sensitivity  

DEFF Research Database (Denmark)

In the Danish serological surveillance for Salmonella in poultry (serum and egg yolk) a mix-ELISA is used, based on S. typhimurium and S. enteritidis antigens (Feld et al., 2000). When we evaluated results of the test retrospectively, over the years an unacceptably large fraction of seropositive findings could not be confirmed by the subsequent confirmatory bacteriological sampling in the herd. Therefore we tried to enhance specificity of the ELISA, without losing sensitivity, by refining the antigens used.

Lauritsen, Klara TØlbØl; Lind, Peter

2014-01-01

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Refining the LPS-antigen in Salmonella antibody ELISA for poultry enhanced specificity without impairing sensitivity.  

DEFF Research Database (Denmark)

In the Danish serological surveillance for Salmonella in poultry (serum and egg yolk) an indirect mix - ELISA is used, based on S. typhimurium and S. enteritidis antigens (Feld et al., 2000). When we evaluated results of the test retrospectively, over the years an unacceptably large fraction of seropositive findings could not be confirmed by the subsequent confirmatory bacteriological sampling in the herd. Therefore we tried to enhance specificity of the ELISA, without losing sensitivity, by refining the antigens used.

Lauritsen, Klara TØlbØl; Klausen, Joan

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Standardization of a quantification method for Salmonella spp. and Shigella spp. in specific liquid media  

Directory of Open Access Journals (Sweden)

Full Text Available Introduction: Chlorination is the most widely used disinfection process for drinking water production. The formation of chlorination carcinogenic by-products and chlorine intoxication by direct manipulation in small communities has motivated the study of alternative disinfection processes. In this sense, processes of advanced oxidation (PAOs have yielded promising results. Escherichia coli (E. coli is customarily used as faecal bacterial indicator to determine the efficiency of disinfection processes. However, it has been shown that E. coli is less resistant to disinfection than other enteric bacteria such as Shigella spp. and Salmonella spp. Additionally, the viable non-culturable (VNC state yields bacteria which are not detectable on many culture media.Objective: The main objective is to standardize a method for counting Salmonella spp. and Shigella spp. in specific liquid media to reliably quantify the bacteriological potential risk related to disinfection processes based on PAO.Methods: The study followed a randomized bi-factorial experimental design and the Duncan multiple comparison test. This design allowed the selection of specific liquid media to fittingly standardize the counting of Salmonella spp. and Shigella spp.Results: We found that the best broth for counting Salmonella typhimurium strain at different concentrations in pure and mixed cultures was the Rappaport broth RP, the EE broth also allowed growing the two bacterial species tested in this research. Nonetheless, the latter results suggest the use of additional tests for this particular broth.Discussion: There was a variation in the counting results when pure cultures were used compared to those obtained from mixtures of microorganisms. It was also noted that Salmonella typhimurium and Shigella sonnei, were recovered from minimal concentrations in both RP and EE broths, respectively. To some extent, this suggests an additional confirmative method when using the EE® broth.Conclusion: MPN is a rapid and inexpensive method; easy to apply in water and other contaminated environments where counting of Shigella spp. and Salmonella spp. is needed to estimate potential bacteriological risks. The broths selected were able to recover the two bacterial species from densities as low as 10 cells per 100 ml.

Sandra Patricia Rivera

2010-03-01

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Rapid and specific detection of Salmonella spp. in animal feed samples by PCR after culture enrichment  

DEFF Research Database (Denmark)

A PCR procedure has been developed for routine analysis of viable Salmonella spp. in feed samples. The objective was to develop a simple PCR-compatible enrichment procedure to enable DNA amplification without any sample pretreatment such as DNA extraction or cell lysis. PCR inhibition by 14 different feed samples and natural background flora was circumvented by the use of the DNA polymerase Tth. This DNA polymerase was found to exhibit a high level of resistance to PCR inhibitors present in these feed samples compared to DyNAzyme II, FastStart Taq, Platinum Taq, Pwo, rTth, Taq, and Tfl. The specificity of the Tth assay was confirmed by testing 101 Salmonella and 43 non-Salmonella strains isolated from feed and food samples. A sample preparation method based on culture enrichment in buffered peptone water and DNA amplification with Tth DNA polymerase was developed. The probability of detecting small numbers of salmonellae in feed, in the presence of natural background flora, was accurately determined and foundto follow a logistic regression model. From this model, the probability of detecting I CFU per 25 g of feed in artificially contaminated soy samples was calculated and found to be 0.81. The PCR protocol was evaluated on 155 naturally contaminated feed samples and compared to an established culture-based method, NMKL-71. Eight percent of the samples were positive by PCR, compared with 3% with the conventional method. The reasons for the differences in sensitivity are discussed. Use of this method in the routine analysis of animal feed samples would improve safety in the food chain.

Löfström, Charlotta; Knutsson, R.

2004-01-01

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Expression, purification and immobilization of the intracellular invertase INVA, from Zymomonas mobilis on crystalline cellulose and Nylon-6.  

Science.gov (United States)

This paper presents two immobilization methods for the intracellular invertase (INVA), from Zymomonas mobilis. In the first method, a chimeric protein containing the invertase INVA, fused through its C-terminus to CBDCex from Cellulomonas fimi was expressed in Escherichia coli strain BL21 (DE3). INVA was purified and immobilized on crystalline cellulose (Avicel) by means of affinity, in a single step. No changes were detected in optimal pH and temperature when INVA-CBD was immobilized on Avicel, where values of 5.5 and 30 degrees C, respectively, were registered. The kinetic parameters of the INVA-CBD fusion protein were determined in both its free form and when immobilized on Avicel. Km and Vmax were affected with immobilization, since both showed an increase of up to threefold. Additionally, we found that subsequent to immobilization, the INVA-CBD fusion protein was 39% more susceptible to substrate inhibition than INVA-CBD in its free form. The second method of immobilization was achieved by the expression of a 6xHis-tagged invertase purified on Ni-NTA resin, which was then immobilized on Nylon-6 by covalent binding. An optimal pH of 5.5 and a temperature of 30 degrees C were maintained, subsequent to immobilization on Nylon-6 as well as with immobilization on crystalline cellulose. The kinetic parameters relating to Vmax increased up to 5.7-fold, following immobilization, whereas Km increased up to 1.7-fold. The two methods were compared showing that when invertase was immobilized on Nylon-6, its activity was 1.9 times that when immobilized on cellulose for substrate concentrations ranging from 30 to 390 mM of sucrose. PMID:18712537

de Los Angeles Calixto-Romo, María; Santiago-Hernández, José Alejandro; Vallejo-Becerra, Vanessa; Amaya-Delgado, Lorena; del Carmen Montes-Horcasitas, María; Hidalgo-Lara, María Eugenia

2008-11-01

35

Desarrollo y validación de una reacción en cadena de la polimerasa múltiple para la identificación de los serogrupos Salmonella enterica B, C2, D y E de Salmonella enterica / Development and validation of a multiplex polymerase chain reaction for molecular identification of Salmonella enterica serogroups B, C2, D and E  

Scientific Electronic Library Online (English)

Full Text Available SciELO Colombia | Language: Spanish Abstract in spanish Introducción. El esquema Kaufmann-White para la serotipificación de Salmonella, reconoce 46 antígenos O y 119 antígenos H, los cuales han permitido la caracterización de 2.541 serovares. La serotipificación es una herramienta epidemiológica útil en la identificación de serovares circulantes y estudi [...] o de brotes, sin embargo, presenta limitaciones técnicas, de interpretación de resultados y alto costo. Objetivo. Desarrollar una prueba de reacción en cadena de la polimerasa múltiple (PCR-M) como alternativa para identificar los serogrupos B, C2, D y E de Salmonella enterica. Materiales y métodos. Se desarrolló una PCR-M para detectar los genes rfbJ de los serogrupos B y C2 y wzx de los serogrupos D y E. Para estandarizar la PCR-M se probaron cepas de referencia de Salmonella pertenecientes a los serogrupos de estudio. Se incluyó el gen invA específico del género Salmonella como control interno de amplificación. La técnica fue validada con un estudio ciego que incluyó 400 aislamientos de Salmonella previamente serotipificados. Resultados. La PCR-M permitió identificar los serogrupos de Salmonella con resultados reproducibles (índice kappa=0,95). La sensibilidad de la prueba estuvo entre 98% y 100% y la especificidad entre 96% y 100%. Conclusiones. El polimorfismo de los genes rfbJ y wzx permitió desarrollar un método de tipificación molecular sensible, específico y reproducible, que podría servir de apoyo a la serotipificación para identificar serogrupos de Salmonella. Abstract in english Introduction. The scheme Kauffman-White (KW) for serotyping of Salmonella recognizes 46 O antigens, and 119 H antigens, thereby permitting the characterization of 2,541 serotypes. The serotyping is a useful epidemiological tool in identifying circulating serotypes and to characterize outbreaks. Howe [...] ver, the method presents technical limitations, difficulty in interpretation of results and high costs. Objective. A multiplex polymerase chain reaction test (M-PCR) was developed as an alternative method for the identification of serogroups B, C2, D, and E of Salmonella enterica. Materials and methods. The M-PCR detected Salmonella genes rfbJ of serogroups B and C2 and wzx of serogroups D and E. To standardize the M-PCR, reference strains of Salmonella serogroups were compared. Amplification of invA gender-specific gene of Salmonella was included as internal control of amplification. To validate the test, a blind study was conducted to identify by M-PCR 400 isolates that had been previously characterized by serology. Results. The M-PCR detected Salmonella serogroups with reproducible results (Kappa index=0.95). The sensitivity of the test was between 98% to 100% and specificity between 96% to 100%. Conclusions. The polymorphisms in the Salmonella genes rfbJ and wzx permitted the development of a method for molecular typing of Salmonella serogroups that was sensitive, specific and reproducible.

Lelia, Lavalett; Miryan Margot, Sánchez; Nélida, Múñoz; Jaime, Moreno; Nora, Cardona-Castro.

36

Desarrollo y validación de una reacción en cadena de la polimerasa múltiple para la identificación de los serogrupos Salmonella enterica B, C2, D y E de Salmonella enterica / Development and validation of a multiplex polymerase chain reaction for molecular identification of Salmonella enterica serogroups B, C2, D and E  

Scientific Electronic Library Online (English)

Full Text Available SciELO Colombia | Language: Spanish Abstract in spanish Introducción. El esquema Kaufmann-White para la serotipificación de Salmonella, reconoce 46 antígenos O y 119 antígenos H, los cuales han permitido la caracterización de 2.541 serovares. La serotipificación es una herramienta epidemiológica útil en la identificación de serovares circulantes y estudi [...] o de brotes, sin embargo, presenta limitaciones técnicas, de interpretación de resultados y alto costo. Objetivo. Desarrollar una prueba de reacción en cadena de la polimerasa múltiple (PCR-M) como alternativa para identificar los serogrupos B, C2, D y E de Salmonella enterica. Materiales y métodos. Se desarrolló una PCR-M para detectar los genes rfbJ de los serogrupos B y C2 y wzx de los serogrupos D y E. Para estandarizar la PCR-M se probaron cepas de referencia de Salmonella pertenecientes a los serogrupos de estudio. Se incluyó el gen invA específico del género Salmonella como control interno de amplificación. La técnica fue validada con un estudio ciego que incluyó 400 aislamientos de Salmonella previamente serotipificados. Resultados. La PCR-M permitió identificar los serogrupos de Salmonella con resultados reproducibles (índice kappa=0,95). La sensibilidad de la prueba estuvo entre 98% y 100% y la especificidad entre 96% y 100%. Conclusiones. El polimorfismo de los genes rfbJ y wzx permitió desarrollar un método de tipificación molecular sensible, específico y reproducible, que podría servir de apoyo a la serotipificación para identificar serogrupos de Salmonella. Abstract in english Introduction. The scheme Kauffman-White (KW) for serotyping of Salmonella recognizes 46 O antigens, and 119 H antigens, thereby permitting the characterization of 2,541 serotypes. The serotyping is a useful epidemiological tool in identifying circulating serotypes and to characterize outbreaks. Howe [...] ver, the method presents technical limitations, difficulty in interpretation of results and high costs. Objective. A multiplex polymerase chain reaction test (M-PCR) was developed as an alternative method for the identification of serogroups B, C2, D, and E of Salmonella enterica. Materials and methods. The M-PCR detected Salmonella genes rfbJ of serogroups B and C2 and wzx of serogroups D and E. To standardize the M-PCR, reference strains of Salmonella serogroups were compared. Amplification of invA gender-specific gene of Salmonella was included as internal control of amplification. To validate the test, a blind study was conducted to identify by M-PCR 400 isolates that had been previously characterized by serology. Results. The M-PCR detected Salmonella serogroups with reproducible results (Kappa index=0.95). The sensitivity of the test was between 98% to 100% and specificity between 96% to 100%. Conclusions. The polymorphisms in the Salmonella genes rfbJ and wzx permitted the development of a method for molecular typing of Salmonella serogroups that was sensitive, specific and reproducible.

Lelia, Lavalett; Miryan Margot, Sánchez; Nélida, Múñoz; Jaime, Moreno; Nora, Cardona-Castro.

2009-06-01

37

A rapid screen of broth enrichments for Salmonella enterica serovars enteritidis, Hadar, Heidelberg, and Typhimurium by Using an allelotyping multiplex PCR that targets O- and H-antigen alleles.  

Science.gov (United States)

Salmonella continues to cause significant foodborne outbreaks, best illustrated with recent outbreaks associated with peanut butter, raw tomatoes, and serrano peppers. To ascertain the likely source of the outbreak, bacterial typing is essential to this process. While PCR has become an important detection tool for pathogens in foods, PCR can also identify strain differences by targeting gene(s) or sequences exhibiting polymorphisms or variability in its distribution within the bacterial population. Over 2,500 Salmonella enterica serovars identified based on antigenic differences in lipopolysaccharide and flagellin have been identified to date. We developed an allelotyping PCR scheme that identifies the O and H alleles associated with S. enterica serovars Enteritidis, Hadar, Heidelberg, Typhimurium, and others, with the same antigen alleles but in different O- and H-allele combinations (e.g., S. enterica Kentucky), and validated it as a screen to identify samples contaminated with these Salmonella serovars. We correctly identified poultry samples containing S. enterica serovars Enteritidis, Kentucky, and Typhimurium from our multiplex screen of primary enrichments of environmental drag swabs. PCR agreed well (kappa values = 0.81 to 1.0) with conventional serotyping methods used to type salmonellae isolated from primary enrichment. Coupled with Salmonella-specific PCR, such as invA, this allelotyping PCR could prove useful in the identification of Salmonella and specific S. enterica serovars present in foods or the environment and could decrease the time and cost associated with conventional serotyping methods. PMID:19833046

Hong, Yang; Liu, Tongrui; Lee, Margie D; Hofacre, Charles L; Maier, Marie; White, David G; Ayers, Sherry; Wang, Lihua; Berghaus, Roy; Maurer, John

2009-10-01

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Artificial Salmonella Vaccines: Salmonella typhimurium O-Antigen-Specific Oligosaccharide-Protein Conjugates Elicit Protective Antibodies in Rabbits and Mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Several saccharides representative of the O-antigenic polysaccharide chain of Salmonella typhimurium (O antigens 4 and 12) were used as haptenic groups covalently linked to bovine serum albumin. The disaccharide abequose 1 ? 3 D-mannose was synthesized, and the [Formula: see text] tetra-, octa- and dodecasaccharides were isolated after cleavage of isolated S. typhimurium O-polysaccharide chains by using bacteriophage endo-glycosidases. Rabbits immunized with the saccharide-protein conjugate...

Svenson, Stefan B.; Lindberg, Alf A.

1981-01-01

39

STRAIN SPECIFIC FEATURES OF THE COMPOSITION AND DYNAMICS OF THE CARBOXYLIC ACIDS DURING CULTIVATION ESCHERICHIA COLI AND SALMONELLA ENTERITIDIS ??????????????????? ??????????? ? ??????? ? ???????? ?????????? ?????? ??? ??????????? ???????? escherichia coli ? salmonella enteritidis  

Directory of Open Access Journals (Sweden)

Full Text Available Qualitative and quantitative composition of the low-molecular carboxylic acids (CA which were contained in the cultural liquid (CL of the four strains of Escherichia coli and Salmonella enteritidis var.Issatchenko was investigated. The differences in the composition of the CA were shown, their dynamics during the bacterial growth and their total concentration in CL were strain specific characteristic. The differences in the composition and dynamics of the CA, which are specific only to probiotic and opportunistic strains were revealed

Polevaya E. V.

2012-03-01

40

Tissue-Specific Salmonella Typhimurium Gene Expression during Persistence in Pigs  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Salmonellosis caused by Salmonella Typhimurium is one of the most important bacterial zoonotic diseases. The bacterium persists in pigs resulting in asymptomatic ‘carrier pigs’, generating a major source for Salmonella contamination of pork. Until now, very little is known concerning the mechanisms used by Salmonella Typhimurium during persistence in pigs. Using in vivo expression technology (IVET), a promoter-trap method based on ?purA attenuation of the parent strain, we identified 37 ...

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

2011-01-01

 
 
 
 
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Antigenic and immunogenic properties of recombinants from Salmonella typhimurium and Salmonella minnesota rough mutants expressing in their lipopolysaccharide a genus-specific chlamydial epitope.  

Science.gov (United States)

Rough mutants from Salmonella typhimurium and Salmonella minnesota were transformed with a plasmid containing a 6.5-kilobase insert of DNA from Chlamydia trachomatis assumed to encode a glycosyltransferase. Transformation resulted in the expression of a genus-specific chlamydial epitope on the lipopolysaccharide (LPS) of the recombinant strains. Proteinase K-digested whole-cell lysates of the recombinants and of controls were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining or Western blot analysis. Two LPS populations were detected in the recombinants, the parent LPS and a faster-migrating component. The latter stained with monoclonal antibody against the genus-specific chlamydial epitope and was not seen in the controls. LPS was extracted and purified from recombinants of S. minnesota R595 and R4 and characterized by the passive hemolysis and passive hemolysis inhibition assays and by hydrolysis kinetics. Different antigenic determinants could be distinguished from each other by the passive hemolysis inhibition test with monospecific antigen-antibody reactions. Rabbits were immunized with heat-killed recombinant bacteria to study the immunogenic properties of the recombinants. In all animals, antibodies were raised against the parent core specificity and against the chlamydia-specific epitope. The data show that the recombinant bacteria are useful as immunogens to prepare polyclonal antisera against chlamydiae and that LPS isolated from them exhibits the same antigenic determinants as chlamydial LPS and may thus be used as a substitute for chlamydial LPS in serological assays. Images PMID:2433222

Brade, L; Nano, F E; Schlecht, S; Schramek, S; Brade, H

1987-01-01

42

Detección de Salmonella spp. en melón Cantaloupe en unidades de producción y unidad de empaque / Detection of Salmonella spp. on Cantaloupe melon production units and packaging facility  

Scientific Electronic Library Online (English)

Full Text Available SciELO Mexico | Language: Spanish Abstract in spanish El melón Cantaloupe (Cucumis melo L.) grupo reticulatus precortado, proveniente del estado de Guerrero, México, se ha asociado con brotes de salmonelosis en Estados Unidos de América y Canadá, por lo que las exportaciones de melón, a estos países, se suspendieron en 2001. En este trabajo se evaluó l [...] a condición sanitaria del melón Cantaloupe, con la detección e identificación de Salmonella, en dos unidades de producción y una unidad de empaque en Zirándaro de los Chávez, Guerrero. Se analizaron 100 melones Cantaloupe (50 de las unidades de producción y 50 de la unidad de empaque), recolectados en enero y abril de 2005, mediante métodos bacteriológicos convencionales y el crecimiento en medios selectivos para la detección de Salmonella, como indicador de contaminación fecal. La proporción de melones con presencia de Salmonella spp. fue 4%, en una de las unidades de producción y 20% en la unidad de empaque. Salmonella se detectó en frutos irrigados con agua de río filtrada pero no clorada y manejados por trabajadores con poca higiene. En pruebas de reacción en cadena de la polimerasa (PCR), dos de seis cepas presuntivas de Salmonella dieron amplificaciones positivas con el par de iniciadores Sal-3 y Sal-4 e invA-1 e invA-2; de las otras cuatro, solo dieron amplificación positiva con invA-1 e invA-2. Estos resultados sugieren que en la región de Zirándaro de los Chávez se tiene más de un serotipo de Salmonella y evidencian la importancia de implementar programas preventivos para asegurar la calidad sanitaria del melón Cantaloupe. Abstract in english Fresh Cantaloupe melons (Cucumis melo L.) group reticulatus coming from the state of Guerrero, Mexico, have been associated with outbreaks of salmonellosis in the United States of America and Canada. These countries suspended the importations of Cantaloupe melon from Mexico due to the outbreaks in 2 [...] 001. This study evaluated the food safety quality of Cantaloupe melon, with the detection and identification of Salmonella in two production units and a packing facility unit in Zirándaro de los Chávez, Guerrero. 100 Cantaloupe melons (50 of the production units and 50 of the packaging unit), collected in January and April 2005, were analyzed by conventional bacteriological methods and growth in selective media for detection of Salmonella, as an indicator of fecal contamination. The proportion of melons with presence of Salmonella was 4%, in one of the field production units and 20% in the packing unit. Salmonella was detected in fruits irrigated with filtered but not chlorinated river water and handled by workers with poor hygiene. Characterization by polymerase chain reaction (PCR) demonstrated that, two of six strains of presumptive Salmonella gave positive amplifications with the pair of primers Sal-3 and Sal-4 as with invA-1and invA-2. For four other isolates only two were observed with invA-1 and invA-2. These results suggest that in the region of Zirándaro de los Chávez there are more than one serotype of Salmonella, and demonstrate the importance of implementing prevention programs to ensure the sanitary quality of Cantaloupe melon.

Lucía, Morales-Hernández; Ana María, Hernández-Anguiano; Cristóbal, Cháidez-Quiroz; Gilberto, Rendón-Sánchez; Trevor, V. Suslow.

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Salmonella typhimurium strain TA100 differentiates several classes of carcinogens and mutagens by base substitution specificity.  

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The mutational specificity of N-methylnitrosourea (MNU), nitrosoguanidine (MNNG), methyl methanesulfonate (MMS), sodium azide (NaN3), 4-nitroquinoline oxide (4NQO), benzo[a]pyrene (BP), nitrofurantoin (NF), aflatoxin B1 (AFB1), adriamycin (ADM) and UVA-activated angelicin in Salmonella typhimurium strain TA100 has been examined using allele-specific oligonucleotide hybridization and DNA sequence analyses. These ten mutagens produced five unique classes of reversion spectra, distinct from spontaneous, or the previously characterized 5-azacytidine, ultraviolet light (UV), 8-methoxypsoralen plus UVA (PUVA) and 60Co-induced mutation spectra. For example, 90% of MNU and MNNG-induced mutations in strain TA100 revertants were G:C-->A:T transitions with the majority (82%) occurring in the first position of the CCC codon. In contrast, NaN3 preferentially induced G:C-->A:T transitions at the second codon position (78%). Although MMS, NQO, BP, NF, ADM and AFB1 induced primarily G:C-->T:A transversions (73-86%), these mutagens fall into two classes based on site preference: NF and AFB1 yielded almost exclusively position two transversions (69-78%) whereas ADM, NQO, BP and MMS exhibited a two-fold preference for site 2 over site 1 (on average 52% versus 22%). Angelicin photomutagenesis resulted in the recovery of G:C-->A:T and G:C-->T:A mutations at both codon positions in roughly equal proportions (approximately 20-25% each). Approximately 1% of the mutagen-induced revertants occurred via extragenic tRNA suppressor mutations, while 1% were multiple (usually tandem double) base substitutions. Ultraviolet mutagenesis experiments demonstrated that tandem base substitutions are promoted by pKM101-encoded mucAB gene products. A comparison of the mutagenic specificity derived for several carcinogens in hisG46 with the responses of several eukaryotic gene targets (e.g. HPRT, aprt, supF) revealed a high concordance between these targets. Thus, the Salmonella hisG46 locus provides a rapid, simple system for determining base substitution specificity and for studying mechanisms of mutagenesis. PMID:8293552

Koch, W H; Henrikson, E N; Kupchella, E; Cebula, T A

1994-01-01

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Desarrollo y validación de una reacción en cadena de la polimerasa múltiple para la identificación de los serogrupos B, C2, D y E de Salmonella enterica Development and validation of a multiplex polymerase chain reaction for molecular identification of Salmonella enterica serogroups B, C2, D and E  

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Full Text Available

Introducción. El esquema Kaufmann-White para la serotipificación de Salmonella, reconoce 46 antígenos O y 119 antígenos H, los cuales han permitido la caracterización de 2.541 serovares. La serotipificación es una herramienta epidemiológica útil en la identificación de serovares circulantes y estudio de brotes, sin embargo, presenta limitaciones técnicas, de interpretación de resultados y alto costo.
Objetivo. Desarrollar una prueba de reacción en cadena de la polimerasa múltiple (PCR-M como alternativa para identificar los serogrupos B, C2, D y E de Salmonella enterica.
Materiales y métodos. Se desarrolló una PCR-M para detectar los genes rfbJ de los serogrupos B y C2 y wzx de los serogrupos D y E. Para estandarizar la PCR-M se probaron cepas de referencia de Salmonella pertenecientes a los serogrupos de estudio. Se incluyó el gen invA específico del género Salmonella como control interno de amplificación. La técnica fue validada con un estudio ciego que incluyó 400 aislamientos de Salmonella previamente serotipificados.
Resultados. La PCR-M permitió identificar los serogrupos de Salmonella con resultados reproducibles (índice kappa=0,95. La sensibilidad de la prueba estuvo entre 98% y 100% y la especificidad entre 96% y 100%.
Conclusiones. El polimorfismo de los genes rfbJ y wzx permitió desarrollar un método de tipificación molecular sensible, específico y reproducible, que podría servir de apoyo a la serotipificación para identificar serogrupos de Salmonella.

Introduction. The scheme Kauffman-White (KW for serotyping of Salmonella recognizes 46 O antigens, and 119 H antigens, thereby permitting the characterization of 2,541 serotypes. The serotyping is a useful epidemiological tool in identifying circulating serotypes and to characterize outbreaks. However, the method presents technical limitations, difficulty in interpretation of results and high costs.
Objective. A multiplex polymerase chain reaction test (M-PCR was developed as an alternative method for the identification of serogroups B, C2, D, and E of Salmonella enterica.
Materials and methods. The M-PCR detected Salmonella genes rfbJ of serogroups B and C2 and wzx of serogroups D and E. To standardize the M-PCR, reference strains of Salmonella serogroups were compared. Amplification of invA gender-specific gene of Salmonella was included as internal control of amplification. To validate the test, a blind study was conducted to identify by M-PCR 400 isolates that had been previously characterized by serology.
Results. The M-PCR detected Salmonella serogroups with reproducible results (Kappa index=0.95. The sensitivity of the test was between 98% to 100% and specificity between 96% to 100%.
Conclusions. The polymorphisms in the Salmonella genes rfbJ and wzx permitted the development of a method for molecular typing of Salmonella serogroups that was sensitive, specific and reproducible.

Jaime Moreno

2009-06-01

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Detection of Salmonella strain by rapid-cycle multiplex PCR  

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Full Text Available Introduction and objective: Salmonellosis is responsible for large numbers of infections in both humans and animals. Conventional methods of isolation of Salmonella strains take 4-7 days to complete and are therefore laborious and require substantial manpower. Our main objective was to develop a rapid detection method using shortened PCR cycles in a conventional thermal cyclers and fast electrophoresis for Salmonella.Materials and methods: The PCR primers for tyv (rfbE, prt (rfbS and invA, genes were used for the rapid identification of S. enterica serovars typhi and paratyphi A, with rapid and short cycles of multiplex PCR. By using very fast and simple DNA extraction method in 10mins, rapid PCR cycles with total times of 35mins and rapid electrophoresis procedure with simple and very cheap buffer in 15mins we were able to separate the PCR products. Results: All references and clinical isolates of Salmonella serovars typhi and paratyphi were accurately identified. Specificity analysis revealed no cross reaction with other enterobacterial strains. The sensitivity of the assay was 1-10 cells. The total time of multiplex PCR from sample preparation to final result is 45 to 50mins.Conclusion: These data indicate that the specificity and sensitivity of optimized rapid cycle multiplex PCR is a potentially valuable tool for rapid diagnosis of S. typhi using a conventional thermal cycler. This method cut the time of a PCR reaction from 3.5h to less than 60mins. These findings could also be applied to other PCR programs detecting various genes allowing researchers to significantly shorten their PCR reaction times.

Fateme Pourali

2011-04-01

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Specific discrimination of three pathogenic Salmonella enterica subsp. enterica serotypes by carB-based oligonucleotide microarray.  

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It is important to rapidly and selectively detect and analyze pathogenic Salmonella enterica subsp. enterica in contaminated food to reduce the morbidity and mortality of Salmonella infection and to guarantee food safety. In the present work, we developed an oligonucleotide microarray containing duplicate specific capture probes based on the carB gene, which encodes the carbamoyl phosphate synthetase large subunit, as a competent biomarker evaluated by genetic analysis to selectively and efficiently detect and discriminate three S. enterica subsp. enterica serotypes: Choleraesuis, Enteritidis, and Typhimurium. Using the developed microarray system, three serotype targets were successfully analyzed in a range as low as 1.6 to 3.1 nM and were specifically discriminated from each other without nonspecific signals. In addition, the constructed microarray did not have cross-reactivity with other common pathogenic bacteria and even enabled the clear discrimination of the target Salmonella serotype from a bacterial mixture. Therefore, these results demonstrated that our novel carB-based oligonucleotide microarray can be used as an effective and specific detection system for S. enterica subsp. enterica serotypes. PMID:24185846

Shin, Hwa Hui; Hwang, Byeong Hee; Seo, Jeong Hyun; Cha, Hyung Joon

2014-01-01

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Host specificity of Salmonella infection in chickens and mice is expressed in vivo primarily at the level of the reticuloendothelial system.  

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By experimental infection, host-specific Salmonella serotypes were shown to demonstrate specificities for chickens, mice, and other laboratory animals. Following oral inoculation, four strains of Salmonella gallinarum and two S. pullorum strains, isolated from diseased poultry, were more virulent for chickens than for mice. By contrast, four strains each of S. choleraesuis and S. dublin, isolated from diseased pigs and cattle, respectively, were more virulent for mice than for chickens. These...

Barrow, P. A.; Huggins, M. B.; Lovell, M. A.

1994-01-01

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Application of an Impedimetric Technique for the Detection of Lytic Infection of Salmonella spp. by Specific Phages  

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Full Text Available This study was performed to evaluate the adaption of the impedimetric method to detect the lytic infection by Salmonella-specific bacteriophages and to provide a higher selectivity to this rapid method in detecting Salmonella spp. by using specific agents. Three bacteriophages and twelve strains of Salmonella spp. were tested. Each of the twelve strains was used separately to inoculate TSB together with each one of the phages. The inoculum concentration was between 106 and 107?cfu/mL, at a cell: phage ratio of 1 : 100. From the sample analysis, based on conductance (G measurements (37°C, the infection could be detected, by observation of both detection-time delay and distinct curve trends. The main conclusions were that kinetic detection by impedance microbiology with phage typing constitutes a method of determining whether a test microorganism is sensitive to the bacteriophage and a method to evaluate whether a lytic bacteriophage is present in a sample, by affecting bacterial growth rate/metabolic change.

Paula C. Teixeira

2009-01-01

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Purification of O-specific polysaccharide from lipopolysaccharide produced by Salmonella enterica serovar Paratyphi A.  

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The O specific polysaccharide (OSP) of the lipopolysaccharide (LPS) of Salmonella enterica serovar Paratyphi A is a protective antigen and the target for vaccine development. LPS is the major constituent of the outer membrane of S. Paratyphi A with the OSP exposed on the surface, in addition to the cell associated LPS a large amount of free LPS was present in the fermentation broth. A purification method was developed to take advantage of both sources of LPS and to maximize recovery of OSP. After fermentation the bacterial cells were concentrated and washed, the permeate containing the free LPS was processed separately from the cells. The free LPS was concentrated and washed on a 100kD ultrafiltration membrane to remove low molecular weight impurities. The LPS was then detoxified by separation of the lipid A from the OSP using acid hydrolysis at 100°C, the precipitated lipid A was removed by 0.2?m membrane filtration. Contaminants were then removed by acid precipitation in the presence of sodium deoxycholate. The OSP was concentrated and washed with 1M NaCl then water using a 10kD ultrafiltration membrane then sterile filtered through a 0.2?m membrane filter. The cells were treated by acid hydrolysis at 100°C, the remaining cells, cell debris and precipitate was removed by centrifugation. The filtrate was then treated in the same way as described above for the free LPS. The combined yield of purified OSP from free LPS plus the cells was greater than 880mg/L of culture broth. The method developed yields large amounts of OSP, is scalable and compatible with cGMP so would be readily transferrable to developing country vaccine manufacturers for low cost production of vaccine against S. Paratyphi A. PMID:24631090

Kothari, Sudeep; Kim, Jeong-Ah; Kothari, Neha; Jones, Christopher; Choe, Woo Seok; Carbis, Rodney

2014-05-01

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Molecular characterization of Salmonella strains in individuals with acute diarrhea syndrome in the State of Sucre, Venezuela / Caracterização molecular de cepas de Salmonella em indivíduos com síndrome da diarreia aguda no Estado de Sucre, Venezuela  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese INTRODUÇÃO: Na Venezuela, síndrome da diarreia aguda (SDA) é a principal causa de mórbi-mortalidade, muitas vezes envolvem o gênero Salmonella. Infecções por Salmonella são associadas com gastroenterite aguda, uma das mais comuns intoxicações alimentares causada pelo consumo de água e alimentos cont [...] aminados, principalmente carne. MÉTODOS: Métodos convencionais e moleculares foram usados para detectar cepas de Salmonella em 330 amostras de fezes de indivíduos com SDA de diferentes idades e ambos os sexos. A reação em cadeia da polimerase (PCR) foi utilizada para a caracterização molecular de genes Salmonella invA, sefA e fliC para identificar o gênero e os sorotipos Enteritidis e Typhimurium, respectivamente. RESULTADOS: A maior frequência de indivíduos com SDA foi encontrada em crianças de 0-2 (39,4%) anos, e a frequência total de culturas de fezes positiva foi de 76,9%. Um total de 14 (4,2%) cepas foram bioquímica e imunologicamente identificados como Salmonella enterica subsp. enterica, dos quais 7 foram classificados como pertencentes ao sorotipo Enteritidis, Typhimurium sorotipo 4 e 3 para outros sorotipos. Cepas S. enterica foram distribuídas mais frequentemente em grupos de 3-4 e 9-10 anos de idade. CONCLUSÕES: O método de caracterização molecular usada provou ser altamente específico para tipificar as estirpes dos S. enterica usando tanto DNA extraído de colônias isoladas e direta e caldos de enriquecimento seletivo inoculados com amostras fecais, o que representa uma ferramenta complementar para a detecção e identificação de bactérias que causam a SDA. Abstract in english INTRODUCTION:In Venezuela, acute diarrheic syndrome (ADS) is a primary cause of morbi-mortality, often involving the Salmonella genus. Salmonella infections are associated with acute gastroenteritis, one of the most common alimentary intoxications, and caused by the consumption of contaminated water [...] and food, especially meat. METHODS: Conventional and molecular methods were used to detect Salmonella strains from 330 fecal samples from individuals of different ages and both sexes with ADS. Polymerase chain reaction (PCR) was used for the molecular characterization of Salmonella, using invA, sefA, and fliC genes for the identification of this genus and the serotypes Enteritidis and Typhimurium, respectively. RESULTS: The highest frequency of individuals with ADS was found in children 0-2 years old (39.4%), and the overall frequency of positive coprocultures was 76.9%. A total of 14 (4.2%) strains were biochemically and immunologically identified as Salmonella enterica subsp. enterica, of which 7 were classified as belonging to the Enteritidis serotype, 4 to the Typhimurium serotype, and 3 to other serotypes. The S. enterica strains were distributed more frequently in the age groups 3-4 and 9-10 years old. CONCLUSIONS: The molecular characterization method used proved to be highly specific for the typing of S. enterica strains using DNA extracted from both the isolated colonies and selective enrichment broths directly inoculated with fecal samples, thus representing a complementary tool for the detection and identification of ADS-causing bacteria.

Hectorina, Rodulfo; Marcos De, Donato; Jesús, Luiggi; Elvia, Michelli; Adriana, Millán; Miriam, Michelli.

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Biomagnetic separation of Salmonella Typhimurium with high affine and specific ligand peptides isolated by phage display technique  

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Analyses of food-borne pathogens are of great importance in order to minimize the health risk for customers. Thus, very sensitive and rapid detection methods are required. Current conventional culture techniques are very time consuming. Modern immunoassays and biochemical analysis also require pre-enrichment steps resulting in a turnaround time of at least 24 h. Biomagnetic separation (BMS) is a promising more rapid method. In this study we describe the isolation of high affine and specific peptides from a phage-peptide library, which combined with BMS allows the detection of Salmonella spp. with a similar sensitivity as that of immunomagnetic separation using antibodies.

Steingroewer, Juliane; Bley, Thomas; Bergemann, Christian; Boschke, Elke

2007-04-01

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Development and evaluation of a multiplex real-time polymerase chain reaction procedure to clinically type prevalent Salmonella enterica serovars.  

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A multiplex real-time polymerase chain reaction procedure was developed to identify the most prevalent clinical isolates of Salmonella enterica subsp. enterica. Genes from the rfb, fliC, fljB, and viaB groups that encode the O, H, and Vi antigens were used to design 15 primer pairs and TaqMan probes specific for the genes rfbJ, wzx, fliC, fljB, wcdB, the sdf-l sequence, and invA, which was used as an internal amplification control. The primers and probes were variously combined into six sets. The first round of reactions used two of these sets to detect Salmonella O:4, O:9, O:7, O:8, and O:3,10 serogroups. Once the serogroups were identified, the results of a second round of reactions that used primers and probes for the flagellar antigen l genes, 1,2; e,h; g,m; d; e,n,x; and z(10), and the Vi gene were used to identify individual serovars. The procedure was standardized using 18 Salmonella reference strains and other enterobacteria. The procedure's reliability and sensitivity was evaluated using 267 randomly chosen serotyped Salmonella clinical isolates. The procedure had a sensitivity of 95.5% and was 100% specific. Thus, our technique is a quick, sensitive, reliable, and specific means of identifying S. enterica serovars and can be used in conjunction with traditional serotyping. Other primer and probe combinations could be used to increase the number of identifiable serovars. PMID:20110454

Muñoz, Nélida; Diaz-Osorio, Miguel; Moreno, Jaime; Sánchez-Jiménez, Miryan; Cardona-Castro, Nora

2010-03-01

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Preparation and evaluation of immunogenic conjugates of Salmonella enterica serovar Typhi O-specific polysaccharides with diphtheria toxoid.  

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Typhoid fever, caused by Salmonella enterica serovar Typhi (S. Typhi), is a major health problem particularly in developing countries. The available vaccines have certain limitations regarding their efficacy, and inability to induce an immune response especially in individuals under 2 years of age. Conjugate vaccines which consist of a bacteria-specific polysaccharide chemically bound to a carrier protein overcome these problems by inducing a T-cell dependent immune response characterized by enhanced immunogenicity in all ages. In this study, O-specific polysaccharides (OSP) of S. Typhi were conjugated to diphtheria toxoid (DT) using adipic acid dihydrazide (ADH) as a linker. These conjugates (OSP-AH-DT) were then evaluated for their immunogenicity using mice as a model and showed significantly higher levels of IgG ELISA titers (P = 0.0241 and 0.0245) than lipopolysaccharides alone. Different immunization  schedules were compared and it was found that schedule-B (three injections with 4-weeks interval) induced higher immune responses than schedule-A (three injections with 2-weeks interval). We showed that diphtheria toxoid can be successfully employed as a carrier protein for conjugation with Salmonella OSP and play an important role in facilitating adequate immune response. PMID:22426380

Ali, Aamir; An, So Jung; Cui, Changfa; Haque, Abdul; Carbis, Rodney

2012-02-01

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Salmonella typhimurium's transthyretin-like protein is a host-specific factor important in fecal survival in chickens.  

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The transthyretin-like protein (TLP) from Salmonella enterica subspecies I is a periplasmic protein with high level structural similarity to a protein found in mammals and fish. In humans, the protein homologue, transthyretin, binds and carries retinol and thyroxine, and a series of other, unrelated aromatic compounds. Here we show that the amino acid sequence of the TLP from different species, subspecies and serovars of the Salmonella genus is highly conserved and demonstrate that the TLP gene is constitutively expressed in S. Typhimurium and that copper and other divalent metal ions severely inhibit enzyme activity of the TLP, a cyclic amidohydrolase that hydrolyses 5-hydroxyisourate (5-HIU). In order to determine the in vivo role of the S. Typhimurium TLP, we constructed a strain of mouse-virulent S. Typhimurium SL1344 bearing a mutation in the TLP gene (SL1344 ?yedX). We assessed the virulence of this strain via oral inoculation of mice and chickens. Whilst SL1344 ?yedX induced a systemic infection in both organisms, the bacterial load detected in the faeces of infected chickens was significantly reduced when compared to the load of S. Typhimurium SL1344. These data demonstrate that the TLP gene is required for survival of S. Typhimurium in a high uric acid environment such as chicken faeces, and that metabolic traits of Salmonellae in natural and contrived hosts may be fundamentally different. Our data also highlight the importance of using appropriate animal models for the study of bacterial pathogenesis especially where host-specific virulence factors or traits are the subject of the study. PMID:23284609

Hennebry, Sarah C; Sait, Leanne C; Mantena, Raju; Humphrey, Thomas J; Yang, Ji; Scott, Timothy; Kupz, Andreas; Richardson, Samantha J; Strugnell, Richard A

2012-01-01

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Proline transport in Salmonella typhimurium: putP permease mutants with altered substrate specificity.  

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The putP gene encodes a proline permease required for Salmonella typhimurium LT2 to grow on proline as the sole source of nitrogen. The wild-type strain is sensitive to two toxic proline analogs (azetidine-2-carboxylic acid and 3,4-dehydroproline) also transported by the putP permease. Most mutations in putP prevent transport of all three substrates. Such mutants are unable to grow on proline and are resistant to both of the analogs. To define domains of the putP gene that specify the substra...

Dila, D. K.; Maloy, S. R.

1986-01-01

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Spontaneous excision of the Salmonella enterica serovar Enteritidis-specific defective prophage-like element phiSE14.  

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Salmonella enterica serovar Enteritidis has emerged as a major health problem worldwide in the last few decades. DNA loci unique to S. Enteritidis can provide markers for detection of this pathogen and may reveal pathogenic mechanisms restricted to this serovar. An in silico comparison of 16 Salmonella genomic sequences revealed the presence of an approximately 12.5-kb genomic island (GEI) specific to the sequenced S. Enteritidis strain NCTC13349. The GEI is inserted at the 5' end of gene ydaO (SEN1377), is flanked by 308-bp imperfect direct repeats (attL and attR), and includes 21 open reading frames (SEN1378 to SEN1398), encoding primarily phage-related proteins. Accordingly, this GEI has been annotated as the defective prophage SE14 in the genome of strain NCTC13349. The genetic structure and location of phiSE14 are conserved in 99 of 103 wild-type strains of S. Enteritidis studied here, including reference strains NCTC13349 and LK5. Notably, an extrachromosomal circular form of phiSE14 was detected in every strain carrying this island. The presence of attP sites in the circular forms detected in NCTC13349 and LK5 was confirmed. In addition, we observed spontaneous loss of a tetRA-tagged version of phiSE14, leaving an empty attB site in the genome of strain NCTC13349. Collectively, these results demonstrate that phiSE14 is an unstable genetic element that undergoes spontaneous excision under standard growth conditions. An internal fragment of phiSE14 designated Sdf I has been used as a serovar-specific genetic marker in PCR-based detection systems and as a tool to determine S. Enteritidis levels in experimental infections. The instability of this region may require a reassessment of its suitability for such applications. PMID:20172996

Santiviago, Carlos A; Blondel, Carlos J; Quezada, Carolina P; Silva, Cecilia A; Tobar, Pia M; Porwollik, Steffen; McClelland, Michael; Andrews-Polymenis, Helene L; Toro, Cecilia S; Zaldívar, Mercedes; Contreras, Inés

2010-04-01

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

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

Chin'ombe Nyasha

2009-04-01

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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.  

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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 class 1 integrons carrying gene cassettes conferring resistance mainly to aminoglycosides are widespread among the MDR Salmonella serovars isolated from humans in Egypt, indicating the important role of these genetic elements in the dissemination of multidrug resistance. PMID:24535570

Osman, K M; Hassan, W M M; Mohamed, R A H

2014-08-01

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Studies on the substrate specificity of a GDP-mannose pyrophosphorylase from Salmonella enterica  

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Full Text Available A series of methoxy and deoxy derivatives of mannopyranose-1-phosphate (Manp-1P were chemically synthesized, and their ability to be converted into the corresponding guanosine diphosphate mannopyranose (GDP-Manp analogues by a pyrophosphorylase (GDP-ManPP from Salmonella enterica was studied. Evaluation of methoxy analogues demonstrated that GDP-ManPP is intolerant of bulky substituents at the C-2, C-3, and C-4 positions, in turn suggesting that these positions are buried inside the enzyme active site. Additionally, both the 6-methoxy and 6-deoxy Manp-1P derivatives are good or moderate substrates for GDP-ManPP, thus indicating that the C-6 hydroxy group of the Manp-1P substrate is not required for binding to the enzyme. When taken into consideration with other previously published work, it appears that this enzyme has potential utility for the chemoenzymatic synthesis of GDP-Manp analogues, which are useful probes for studying enzymes that employ this sugar nucleotide as a substrate.

Lu Zou

2012-08-01

60

Visualization of specific gene expression in individual Salmonella typhimurium cells by in situ PCR  

DEFF Research Database (Denmark)

An in situ PCR protocol by which we can monitor the presence or absence of lac mRNA in individual cells of a Salmonella typhimurium F' lac(+) strain has been developed. In this protocol, fixed cells are permeabilized with lysozyme and subjected to a seminested reverse transcriptase PCR using reporter molecule-labeled primers, and subsequently, intracellular reporter molecules are detected microscopically at the individual-cell level by use of a horseradish peroxidase-conjugated antifluorescein antibody assay. In order to determine the sensitivity of the in situ PCR assay, the ability to detect lac mRNA in suboptimally isopropyl-beta-D-thiogalactopyranoside-induced cells was investigated. By use of a single-cell beta-galactosidase assay, it was confirmed that homogeneous suboptimally induced cultures of S. typhimurium F' lacY cells could be established, and the number of functional lac mRNAs in individual cells was estimated from standard population level beta-galactosidase assays, Cells estimated to contain asingle lac mRNA were detected as containing lac mRNA by the in situ PCR method. Conclusively, we demonstrate the potential of in situ PCR for detection of even poorly expressed mRNA in individual bacterial cells.

Tolker-Nielsen, Tim; Molin, SØren

1997-01-01

 
 
 
 
61

Chimeric flagellin expressed by Salmonella typhimurium induces an ESAT-6-specific Th1-type immune response and CTL effects following intranasal immunization  

Science.gov (United States)

The flagellin component FliC of Salmonella typhimurium is capable of activating the innate immune system via specific interactions with TLR5 and can also act as a carrier of foreign antigen to elicit antigen-specific immune responses. Thus, we constructed an attenuated Salmonella strain SL5928(fliC/esat) expressing chimeric flagellin that contained the ESAT-6 antigen coding sequence of Mycobacterium tuberculosis inserted into the highly variable region of the Salmonella flagellin coding gene fliCi. The chimeric flagellin functioned normally, as demonstrated using a flagella swarming assay and electron microscopy. To analyze the effects of chimeric flagellin, the cell-mediated immune response and cytotoxic T lymphocyte (CTL) effects specific for ESAT-6 antigen were tested after intranasal immunization of mice with flagellated Salmonella SL5928(fliC/esat). The results showed that SL5928(fliC/esat) intranasal immunization can strongly elicit an ESAT-6-specific T helper (Th) 1-type immune response in mucosal lymphoid tissues, such as nasopharynx-associated lymph nodes, lung and Peyer's patches, and a Th1/Th2 response was elicited in spleen and mesenteric lymph nodes. Furthermore, intranasal immunization of SL5928(fliC/esat) produced efficient CTL effects, as demonstrated using a 5- and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) assay. Thus, our study revealed that Salmonella flagellin acts as a carrier for foreign antigen and triggers strong Th1 and CTL responses during intranasal immunization. Chimeric flagellin is potentially an effective strategy for the development of novel vaccines against tuberculosis in humans and animals. PMID:21841816

Zhang, Hui; Liu, Liu; Wen, Ke; Huang, Jinlin; Geng, Shizhong; Shen, Junsong; Pan, Zhiming; Jiao, Xinan

2011-01-01

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[The value of PCR method in the species-level identification of a mucoid Salmonella sp. strain isolated from the urine culture of a case with asymptomatic nephrolithiasis].  

Science.gov (United States)

Colonies of the Salmonella strains usually show a smooth (S) character. Therefore, Salmonella strains producing mucoid colony are very rarely encountered in the literature. Identification of the mucoid Salmonella strains to the species level is difficult via conventional methods, since the mucus layer does not allow the bacterium to respond to the antigenic reactions. In this study we aimed to emphasize the identification of Salmonella serotypes by the polymerase chain reaction (PCR) when rough (R) or mucoid (M) Salmonella isolates are encountered in the laboratory. The urine culture of a 17-year-old female patient revealed growth of 100.000 cfu/mL gram-negative bacilli in mucoid colony morphology. The isolate was identified as Salmonella sp. by biochemical tests and Vitek 2 (bioMérieux, France) automated identification system. Agglutination tests showed negative reaction with the known antiserums. Absence of agglutination was attributed to the mucoid character of the isolate. Identification of the Salmonella sp. was confirmed by Vitek MS MALDI-TOF (bioMérieux, France) analysis method, however, the serotype of the strain could not be identified. In order to verify that the mucoid colony was Salmonella spp., species-specific PCR was performed using invA primers, and Salmonella sp. identification was verified by observing a 284 base-pair (bp) PCR amplicon. Subsequently, serogrouping was done by multiplex-PCR (mPCR), which could identify the O:B (O:4), O:C1 (O:7), O:C2-C3 (O:8), O:D (O:9, O:9,46, O:9,46,27), and O:E (O:3,10, O:3,19) somatic antigens. It was detected that the mucoid Salmonella sp. formed a band of approximately 615 bp in size and took place in group D. Another mPCR directed towards O:D1(O:9) and O:E1(3,10) somatic antigens to detect subgroups of group D mucoid Salmonella spp., revealed that the isolate formed a DNA band of approximately 624 bp in size and took place in group D1 which is usually isolated from human. Modified version of another mPCR was used to determine phase-1 flagellar antigen of common Salmonella serovars, as well as to determine the phase-1 flagellar antigen of mucoid Salmonella spp. in group D1. Thus, the isolate was serotyped as Salmonella Enteritidis (1.9,12:g,m:-). Antibiotic susceptibility test performed by disc diffusion method in line with the recommendations of CLSI, revealed that the isolate was susceptible to ampicillin, ciprofloxacin, ceftriaxone, trimethoprim-sulfamethoxazole and chloramphenicol. In conclusion, PCR is a reliable and rapid alternative method that contributes to the conventional serotyping of Salmonella when rough or mucoid strains that lack somatic and flagellar antigens, are isolated. PMID:24506726

Bay?nd?r Bilman, Fulya; Günayd?n, Elçin; Turhano?lu, Mine; Akkoç, Ali

2014-01-01

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The impact of a specific blend of essential oil components and sodium butyrate in feed on growth performance and Salmonella counts in experimentally challenged broilers.  

Science.gov (United States)

Essential oils (EO) and short-chain fatty acids have potential antimicrobial activity in broilers. This study aimed to investigate the effect of a specific blend of EO and a combination of this blend of EO with sodium-butyrate on growth performance and Salmonella colonization in broilers. A total of 480 one-day-old male broilers were distributed into 5 treatments (8 pens per treatment and 12 birds per pen) and reared during 42 d in experimental conditions. Dietary treatments consisted of the addition of different doses of EO (0 mg/kg, control; 50 mg/kg, EO50 and 100 mg/kg, EO100) or a combination of EO with 1 g/kg of sodium-butyrate (B; EO50 + B, EOB50 and EO100 + B, EOB100) to a basal diet. All birds were orally infected with 10(8) cfu of Salmonella Enteritidis on d 7 of study. Individual BW and feed intake per pen were measured at arrival and on a weekly basis. The prevalence and enumeration of Salmonella in feces was determined per treatment at 72 h postinfection and on d 23 and 37 of study. At slaughter, cecal content and liver samples from 16 birds per treatment were cultured for Salmonella and cecal pH was measured. No differences were observed on growth performance among treatments. All fecal samples analyzed were positive for Salmonella from d 10 to the end of the rearing period. At slaughter, Salmonella contamination (positive samples) in cecum was lower in birds fed EOB50 compared with the other treatments (P < 0.05), whereas birds fed the control diet showed the highest colonization rates. The pH of the cecal content was not different among treatments. Thus, EO or its combination with sodium-butyrate did not affect growth performance. However, a clear effectiveness of these products was observed in Salmonella control, especially when low doses of EO were combined with sodium-butyrate (EOB50). PMID:24604853

Cerisuelo, A; Marín, C; Sánchez-Vizcaíno, F; Gómez, E A; de la Fuente, J M; Durán, R; Fernández, C

2014-03-01

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Salmonella typhimurium IroN and FepA Proteins Mediate Uptake of Enterobactin but Differ in Their Specificity for Other Siderophores  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Salmonella typhimurium possesses two outer membrane receptor proteins, IroN and FepA, which have been implicated in the uptake of enterobactin. To determine whether both receptors have identical substrate specificities, fepA and iroN mutants and a double mutant were characterized. While both receptors transported enterobactin, the uptake of corynebactin and myxochelin C was selectively mediated by IroN and FepA, respectively.

Rabsch, Wolfgang; Voigt, Wolfgang; Reissbrodt, Rolf; Tsolis, Rene?e M.; Ba?umler, Andreas J.

1999-01-01

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Comprehensive analysis of Salmonella sequence polymorphisms and development of a LDR-UA assay for the detection and characterization of selected serotypes.  

Science.gov (United States)

Salmonella is a major cause of food-borne disease, and Salmonella enterica subspecies I includes the most clinically relevant serotypes. Salmonella serotype determination is important for the disease etiology assessment and contamination source tracking. This task will be facilitated by the disclosure of Salmonella serotype sequence polymorphisms, here annotated in seven genes (sefA, safA, safC, bigA, invA, fimA, and phsB) from 139 S. enterica strains, of which 109 belonging to 44 serotypes of subsp. I. One hundred nineteen polymorphic sites were scored and associated to single serotypes or to serotype groups belonging to S. enterica subsp. I. A diagnostic tool was constructed based on the Ligation Detection Reaction-Universal Array (LDR-UA) for the detection of polymorphic sites uniquely associated to serotypes of primary interest (Salmonella Hadar, Salmonella Infantis, Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Gallinarum, Salmonella Virchow, and Salmonella Paratyphi B). The implementation of promiscuous probes allowed the diagnosis of ten further serotypes that could be associated to a unique hybridization pattern. Finally, the sensitivity and applicability of the tool was tested on target DNA dilutions and with controlled meat contamination, allowing the detection of one Salmonella CFU in 25 g of meat. PMID:21611799

Lauri, Andrea; Castiglioni, Bianca; Mariani, Paola

2011-07-01

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THE USE OF THE SPECIFIC ANTI-SALMONELLA POLYCLONAL ANTIBODIES ISOLATED FROM HEN EGGS, IN SALMONELLOSIS PROPHYLAXIS  

Directory of Open Access Journals (Sweden)

Full Text Available The administration of increased doses of antibodies in groupsexperimentally infected with Salmonella gallinarum, in order torecord the efficiency of their administration in salmonellosisprophylaxis was the aim of our research. When a low infectiondose, 1x107 CFU Salmonella gallinarum, was used theadministration of IgY polyclonal antibodies as immunoglobulinextract, or even yolk administration had a protective effect againstgerms invasion. This effect was not recorded when a 10 folds higherdose was administered (1x108 CFU. The prophylactic effect of theadministration of polyclonal antibodies is demonstrated.

CRISTE ADRIANA

2007-01-01

67

MUTATION SPECTRA OF GLU-P-1 IN SALMONELLA: INDUCTION OF HOTSPOT FRAMESHIFTS AND SITE-SPECIFIC BASE SUBSTITUTIONS  

Science.gov (United States)

The mutations induced in approximately 2,000 mutants of Salmonella by the heterocyclic@ amine Glu-P-1 were determined by colony probe hybridization and PCR/DNA sequence analysis. ll of the mutations were at sites containing guanine, which is the base at which Glu-P-1 forms DNA ad...

68

Development of an enzyme-linked immunosorbent assay for quantification of Salmonella enteritidis-specific antibodies in egg yolk  

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Full Text Available The present study aims at developing an indirect ELISA to quantify yolk antibodies specific to all surface proteins of the invasive Salmonella Enteritidis (SE, which acquired the 1.8, 14.1, and ~ 50 Kb plasmids. An ELISA checkerboard was used in four different experiments to account for the different parameters included in the preliminary ELISA procedure, and consequently to maximize the difference in Optical Density (OD values between control positive and negative yolk samples. The first experiment aimed at studying the impact of 5% Bovine Serum Albumin (BSA dissolved in distilled water as a blocking reagent on a 28 µg/well SE antigen-coated plate, while applying the positive and negative control yolk samples to different concentrations of Phosphate-Buffered Saline (PBS.Conjugate application was maintained constant at a dilution of 1:500 in PBS. The second experiment was similar to the first one, but the positive and negative control yolk samples were diluted in PBS-Tween 20, and the conjugate dilution was changed to 1:1500 in PBS-Tween 20. In the third experiment, the conjugate was diluted at 1:1500 in 5% BSA/PBS-Tween 20 diluent or PBS-Tween 20 diluent with no 5% BSA. The objective of the fourth experiment was to study the impact of four different concentrations of SE-coated antigen levels (28µg/well, 56µg/well, 84µg/well, and 112µg/well, while fixing the blocking step with 5% BSA in distilled water, and the conjugate dilution set at 1:1000 in 5% BSA/PBSTween 20, and fixing the control yolk samples dilution at 1% in PBS-Tween 20. This last experimental procedure allowed the highest difference in mean absorbance OD values of the positive control minus the negative control samples, which was equivalent to 0.381. In addition, the final protocol for this ELISA was applied on individual egg yolk samples of two groups of chicken layers: one challenged in the esophagus at 11 days with 5.4 x 10(10 CFU/ml/bird of SE, and the second group was not challenged. The mean OD values of the egg yolk of antibodies specific against SE of the two groups were significantly different (0.8578 versus 0.5250; p<0.05, which indicates the possible application of the developed ELISA for screening SE infection by examining egg yolks produced by commercial layers.

IT Tayeb

2006-12-01

69

Development of an enzyme-linked immunosorbent assay for quantification of Salmonella enteritidis-specific antibodies in egg yolk  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english The present study aims at developing an indirect ELISA to quantify yolk antibodies specific to all surface proteins of the invasive Salmonella Enteritidis (SE), which acquired the 1.8, 14.1, and ~ 50 Kb plasmids. An ELISA checkerboard was used in four different experiments to account for the differe [...] nt parameters included in the preliminary ELISA procedure, and consequently to maximize the difference in Optical Density (OD) values between control positive and negative yolk samples. The first experiment aimed at studying the impact of 5% Bovine Serum Albumin (BSA) dissolved in distilled water as a blocking reagent on a 28 µg/well SE antigen-coated plate, while applying the positive and negative control yolk samples to different concentrations of Phosphate-Buffered Saline (PBS).Conjugate application was maintained constant at a dilution of 1:500 in PBS. The second experiment was similar to the first one, but the positive and negative control yolk samples were diluted in PBS-Tween 20, and the conjugate dilution was changed to 1:1500 in PBS-Tween 20. In the third experiment, the conjugate was diluted at 1:1500 in 5% BSA/PBS-Tween 20 diluent or PBS-Tween 20 diluent with no 5% BSA. The objective of the fourth experiment was to study the impact of four different concentrations of SE-coated antigen levels (28µg/well, 56µg/well, 84µg/well, and 112µg/well), while fixing the blocking step with 5% BSA in distilled water, and the conjugate dilution set at 1:1000 in 5% BSA/PBSTween 20, and fixing the control yolk samples dilution at 1% in PBS-Tween 20. This last experimental procedure allowed the highest difference in mean absorbance OD values of the positive control minus the negative control samples, which was equivalent to 0.381. In addition, the final protocol for this ELISA was applied on individual egg yolk samples of two groups of chicken layers: one challenged in the esophagus at 11 days with 5.4 x 10(10) CFU/ml/bird of SE, and the second group was not challenged. The mean OD values of the egg yolk of antibodies specific against SE of the two groups were significantly different (0.8578 versus 0.5250; p

IT, Tayeb; P, Nehme; L, Jaber; EK, Barbour.

2006-12-01

70

A study of Salmonella in pigs from birth to carcass: serotypes, genotypes, antibiotic resistance and virulence profiles.  

Science.gov (United States)

A study was undertaken to investigate Salmonella in pigs at each step from birth to carcass. Environmental and/or pig samples were taken at birth, farrowing, 1st weaning, 2nd weaning, finishing, transport, lairage, bleeding and chilling of carcasses and tested for Salmonella. All isolates were characterised in terms of serotype, phage type (where relevant) and subtyping with pulsed field gel electrophosesis (PFGE). Isolates were tested for antibiotic resistance, resistance (intI1, bla(CIT), bla(Tem), bla(PSE-1), bla(OXA-1), floR, catA1, aadA1, aadA2, tetA, tetB, tetG, sul1and aphA1) and virulence (invA, rck, spvC and pefA) genes. PCR was also performed to test for the presence of the left junction, thdF-S001 and the right junction, S004-int2 or S004-yidY of Salmonella genomic island 1 (SGI1). Overall 4.3%, 27.5% and 5% of environmental, throat/rectal and carcass samples were Salmonella positive, respectively. S. Typhimurium DT193 was detected during production, while S. Typhimurium DT17 and U311 were present in lairage at the abattoir, where strain characterisation suggested cross contamination of the live animals occurred. The carcasses were also cross contaminated with S. Brandenburg during processing. PFGE grouped the isolates by serotype and/or phage type. The DT193 isolates displayed the ACSSuTTmMn/Gm resistance phenotype and carried the invA, spvC, rck, bla-tem, aadA2, tetA, strA virulence/antibiotic resistance markers; U311 showed an ASSuTMn resistance pattern and carried invA and tetB; DT17 was sensitive to all antibiotics tested but invA, spv and rck positive while S. Brandenburg displayed neither resistance nor virulence gene carriage. None of the isolates possessed class 1 integrons and all isolates were negative for the left and right junctions of SGI1. It was concluded that control activities should target improved biosecurity at farm level and better sanitation in lairage. This study also provides further evidence that multiple drug resistance may be associated with non-SGI1 Salmonella strains. The continued emergence of non-DT104 S. Typhimurium isolates exhibiting multidrug resistance is a cause for concern as is the persistence of highly virulent Salmonella strains in the abattoir environment. PMID:23290238

Bolton, Declan J; Ivory, Claire; McDowell, David

2013-01-01

71

Mutation of a Salmonella Serogroup-C1-Specific Gene Abrogates O7-Antigen Biosynthesis and Triggers NaCl-Dependent Motility Deficiency  

Science.gov (United States)

Several molecular detection marker genes specific for a number of individual Salmonella serogroups have been recently identified in our lab by comparative genomics for the genotyping of diverse serogroups. To further understand the correlation between serotype and genotype, the function of a Salmonella serogroup-C1-specific gene (SC_2092) was analyzed in this study. It was indicated from the topological prediction using the deduced amino acid sequence of SC_2092 that this putative protein was highly similar to the confirmed Wzx flippases. Furthermore, SDS-PAGE revealed that lipopolysaccharide (LPS) biosynthesis, specifically O-antigen synthesis, was incomplete in an SC_2092 in-frame deletion mutant, and no agglutination reaction with the O7 antibody was exhibited in this mutant. Therefore, it was revealed that this Salmonella serogroup-C1-specific gene SC_2092 encoded a putative flippase, which was required for O7-polysaccharide biosynthesis, and was designated here as wzxC1. Subsequently, the effects of the deletion of wzxC1 on bacterial motility and sodium chloride (NaCl) tolerance were evaluated. The wzxC1 mutant lacked swarming motility on solid surfaces and was impaired in swimming motility in soft agar. Moreover, microscopic examination and RT-qPCR exhibited that an increased auto-aggregation and a strong defect in flagella expression, respectively, were responsible for the reduced motility in this mutant. In addition, the wzxC1 mutant was more sensitive than the wild-type strain to NaCl, and auto-aggregation of mutant cells was observed immediately up on the addition of 1% NaCl to the medium. Interestingly, the motility deficiency of the mutant strain, as well as the cell agglomeration and the decrease in flagellar expression, were relieved in a NaCl-free medium. This is the first study to experimentally demonstrate a connection between a Salmonella serogroup specific gene identified by comparative genomics with the synthesis of a specific O-antigen biosynthesis. Also, our results show that the mutation of wzxC1 triggers a NaCl-dependent motility deficiency. PMID:25211341

Zhou, Xiujuan; Liu, Bin; Shi, Chunlei; Shi, Xianming

2014-01-01

72

Mutation of a Salmonella serogroup-C1-specific gene abrogates O7-antigen biosynthesis and triggers NaCl-dependent motility deficiency.  

Science.gov (United States)

Several molecular detection marker genes specific for a number of individual Salmonella serogroups have been recently identified in our lab by comparative genomics for the genotyping of diverse serogroups. To further understand the correlation between serotype and genotype, the function of a Salmonella serogroup-C1-specific gene (SC_2092) was analyzed in this study. It was indicated from the topological prediction using the deduced amino acid sequence of SC_2092 that this putative protein was highly similar to the confirmed Wzx flippases. Furthermore, SDS-PAGE revealed that lipopolysaccharide (LPS) biosynthesis, specifically O-antigen synthesis, was incomplete in an SC_2092 in-frame deletion mutant, and no agglutination reaction with the O7 antibody was exhibited in this mutant. Therefore, it was revealed that this Salmonella serogroup-C1-specific gene SC_2092 encoded a putative flippase, which was required for O7-polysaccharide biosynthesis, and was designated here as wzxC1. Subsequently, the effects of the deletion of wzxC1 on bacterial motility and sodium chloride (NaCl) tolerance were evaluated. The wzxC1 mutant lacked swarming motility on solid surfaces and was impaired in swimming motility in soft agar. Moreover, microscopic examination and RT-qPCR exhibited that an increased auto-aggregation and a strong defect in flagella expression, respectively, were responsible for the reduced motility in this mutant. In addition, the wzxC1 mutant was more sensitive than the wild-type strain to NaCl, and auto-aggregation of mutant cells was observed immediately up on the addition of 1% NaCl to the medium. Interestingly, the motility deficiency of the mutant strain, as well as the cell agglomeration and the decrease in flagellar expression, were relieved in a NaCl-free medium. This is the first study to experimentally demonstrate a connection between a Salmonella serogroup specific gene identified by comparative genomics with the synthesis of a specific O-antigen biosynthesis. Also, our results show that the mutation of wzxC1 triggers a NaCl-dependent motility deficiency. PMID:25211341

Zhou, Xiujuan; Liu, Bin; Shi, Chunlei; Shi, Xianming

2014-01-01

73

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

Science.gov (United States)

Development of antibiotic resistance may alter the virulence properties of bacterial organisms. In this study, nine clinical ceftriaxone-susceptible Salmonella enterica serotype Typhimurium strains were subjected to stepwise selection with increasing concentrations of ceftriaxone in culture media. Mutations in virulence-associated genes and antibiotic efflux genes were analyzed by polymerase chain reaction (PCR) and DNA sequencing. The expression levels of virulence genes invA and stn as well as efflux pump genes tolC, arcA, and arcB before and after the selection were measured by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The stepwise selection resulted in the development of Salmonella strains that were highly resistant to ceftriaxone. Sequence analysis did not reveal any mutations or deletions in the examined virulence genes and regulatory gene, but a silent mutation (T423C) in acrR (encoding a repressor for the efflux pump) was detected in most of the ceftriaxone-resistant strains. The qRT-PCR revealed increased expression of the AcrAB-TolC efflux pump and decreased expression of invA and stn in the ceftriaxone-resistant strains. Moreover, decreased invasion into cultured epithelial cells and reduced growth rates were observed with the resistant strains. These results suggest that acquisition of ceftriaxone resistance is associated with the overexpression of the AcrAB-TolC efflux pump and leads to reduced virulence in Salmonella Typhimurium. PMID:23320420

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

2013-01-01

74

A novel multiplex PCR assay for rapid and simultaneous detection of five pathogenic bacteria: Escherichia coli O157:H7, Salmonella, Staphylococcus aureus, Listeria monocytogenes, and Vibrio parahaemolyticus.  

Science.gov (United States)

Rapid and sensitive detection techniques for foodborne pathogens are important to the food industry. However, traditional detection methods rely on bacterial culture in combination with biochemical tests, a process that typically takes 4 to 7 days to complete. Thus, this study was conducted to address the issue of time lag inherent in traditional methods by developing a novel PCR assay for each of five foodborne pathogenic bacteria. This new system consists of a simultaneous screening method using multiplex PCR in a single reaction tube for the rapid and sensitive detection of each of the five bacteria. Specific primers for multiplex PCR amplification of the Shiga-like toxin (verotoxin type II), femA (cytoplasmic protein), toxR (transmembrane DNA binding protein), iap (invasive associative protein), and invA (invasion protein A) genes were designed to allow simultaneous detection of Escherichia coli O157:H7, Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, and Salmonella, respectively. To confirm the specificity of each primer pair for the respective target gene, three types of experiments were carried out using boiled cell lysates and their DNAs. In the multiplex PCR with mixed DNA samples, specific bands for corresponding genes were simultaneously detected from a single reaction. The detection of all five foodborne pathogenic bacteria could be completed in less than 24 h with this novel PCR method. PMID:17685339

Kim, Jeong Soon; Lee, Gang Gweon; Park, Jong Seok; Jung, Yong Hyun; Kwak, Hyo Sun; Kim, Soo Bok; Nam, Young Suk; Kwon, Suk-Tae

2007-07-01

75

Allotypic and idiotypic specificities of anti-Salmonella abortus-equi antibodies produced by rabbits subjected to successive irradiations  

International Nuclear Information System (INIS)

Three out of five rabbits subjected to successive irradiations and immunized against Salmonella abortus-equi produced IgG which, transiently, were not precipitated by anti-a3 anti-allotypic sera, although they carried all the determinants of the a3 allotypic pattern. As a3 IgG from normal rabbits are precipitated by anti-a3 sera, it appeared that the molecular distribution of the a3 allotypic determinants was different on the IgG produced by these irradiated rabbits compared to IgG produced by normal rabbits. After a second irradiation, one of these rabbits produced a high level (50 mg per ml) of anti-Salmonella antibodies of restricted heterogeneity. An anti-idiotypic serum prepared against anti-Salmonella antibodies produced by this rabbit after the first irradiation precipitated the idiotypes it recognised in the serum collected after the first irradiation, while it did not precipitate the idiotypes it recognised in the serum collected after the second irradiation, although they carried all the determinants of the idiotypes of the serum collected after the first irradiation. That probably means that there is a different molecular distribution of the idiotypic determinants between antibodies produced after the first irradiation and antibodies produced after the second irradiation. An antiidiotypic serum prepared against anti-Salmonella antibodies produced after the second irradiation did not distinguish by precipitation in gel medium or by radioimmunoassay between tel medium or by radioimmunoassay between the idiotypes it recognised in antibodies produced after the first irradiation and those in antibodies produced after the second irradiation. The idiotypic similarity thus detected, and the fact that unexpected allotypes were not detected, is in better agreement with the expression of the potentiality of radiation-resistant cells than with the expression of new antibody producing cell clones arising from virgin stem cells. (author)

76

Pathogenicity of Salmonella enteritidis Phage Type 1 Isolate of Malaysia in 21 Day Old Specific-Pathogen Free Chickens  

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Full Text Available Salmonella enteritidis (SE has always been related to subclinical infection in the chickens infected after 2 weeks of hatching. However, few pathogenic phage types were proven for their ability to manifest systemic infection and cause the organism to be shed into the surrounding environment. It was the objective of the study to determine the pathogenicity of SE Phage Type (PT 1 in Specific-Pathogen-Free (SPF chickens. About 93, 21 day old SPF chickens where divided into 3 groups namely the Control, SE and Mortality groups. The chickens were raised separately in caging system and given free access to antibiotic-free ration and water. The SE and Mortality groups were inoculated orally (1.0 mL with SE PT 1 (1x108 cfu mL-1. The chickens in the SE and Control groups were sacrificed at various intervals throughout the trial. Samples were collected for bacterial isolation and histological examination. The mortality percentage of the chickens in the Mortality group was recorded. The study showed that no mortality was recorded throughout the trial in the mortality as well as the SE group. Body weight was lower in the SE group when compared to the Control group throughout the trial except at days 2, 3 and 5 post inoculation (pi reaching its peak at day 14 pi when the SE group body weight was 26% lower than the controls. Clinical signs observed in the SE and Mortality group were represented by diarrhoea, inappetance, ruffled feather and stunted chickens while no abnormal clinical signs where recorded in the Control group. Grossly mild airsacculitis, mild peritonitis and hepatic congestion where recorded in the SE group at day 2 pi until day 5 pi while no gross lesions where recorded in the Control group. SE was first isolated in the caecum (66% at 12 h pi. At day 1 pi SE was isolated from the caecum and spleen (33% whilst at day 2, SE was isolated from the caecum (100% and caecal tonsil (66%. No SE was isolated from the cloacal swabs throughout the trial. The villi height was generally lower in the SE group when compared to the Controls, however it was significantly lower (p<0.05 in the duodenum at 12 h, days 1, 3, 5, 10, 14 and 21 pi; in the jejunum at 6 h, days 2, 14 and 21 pi while in the ileum at days 1, 3 and 5 pi. The crypts depth measurement was fluctuating however it ended up by being higher in the SE group, nevertheless it was significantly lower (p<0.05 in the SE group when compared to the Control group in the duodenum at 6 h and day 14 pi in the jejunum at day 10 pi; in the ileum at 12 h pi. Histopathological changes recorded included hepatitis, congestion and focal areas of necrosis; splenitis, congestion and oedema in the adenoid sheathed arteries; congestion and areas of necrosis in the lymphoi follicles of the bursa of Fabricius; enteritis, congestion and sloughing of necrotic enterocytes in the intestinal villi with presence of bacterial clusters in the villi surface and intestinal lumen. SE rods present in the caecal tonsils were seen to be engulfed by macrophages at days 1 and 2 pi, necrosis of the enterocytes on the villi surface and infiltration of the bacteria was recorded at day 2 pi while at days 5 pi the bacteria multiplication were seen and often located upon the M-like M cells however, no actual engulfment was recorded.

S. Khairani-Bejo

2011-01-01

77

Salmonella Prevention  

Science.gov (United States)

... Administration (FDA) USDA Food Safety and Inspection Service Prevention Quick Tips for Preventing Salmonella Cook poultry, ground ... salmonellosis and many other health problems. More About Prevention There is no vaccine to prevent salmonellosis. Because ...

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Salmonella Yoruba infection in white-tufted-ear marmoset (Callithrix jacchus Infecção por Salmonella Yoruba em sagui-de-tufo-branco (Callithrix jacchus  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study was to describe a fatal salmonellosis case in a non-human female primate (Callithrix jacchus, found in the illegal pet trade in Brazil. The marmoset was sent to the quarantine section of the Guarulhos City Zoo and died in the sequence of an episode of profuse diarrhea. Necropsy findings included mucous enteritis, and liver enlargement and necrosis. Feces and liver fragments were collected for bacteriological tests, which indicated the presence of Salmonella sp.; it was subsequently characterized as pertaining to the Yoruba serotype. The susceptibility profile demonstrated resistance to tetracycline only. The strain was positive for genes that encoded the virulence factors investigated (invA, sefC, pefA and spvC. The results indicated the risk of introduction of Salmonella pathogenic serotypes in primates in captivity.O objetivo deste trabalho foi descrever um caso fatal de salmonelose em uma fêmea primata não humana (Callithrix jacchus, originária de tráfico ilegal no Brasil. O sagui foi enviado para seção de quarentena do Zoológico Municipal de Guarulhos e morreu após um episódio de diarréia profusa. Achados de necropsia incluíram enterite mucosa, hepatomegalia e necrose do fígado. Fezes e fragmentos do fígado foram coletados para testes bacteriológicos e indicaram a presença de Salmonella sp.; subsequentemente caracterizada como sorotipo Yoruba. O perfil de suscetibilidade mostrou resistência somente à tetraciclina. A cepa foi positiva para os genes que codificam os fatores de virulência investigados (invA, sefC, pefA and spvC. Os resultados indicaram o risco de introdução de sorotipos patogênicos de Salmonella por primatas em cativeiro.

Terezinha Knöbl

2011-08-01

79

Salmonella Yoruba infection in white-tufted-ear marmoset (Callithrix jacchus) / Infecção por Salmonella Yoruba em sagui-de-tufo-branco (Callithrix jacchus)  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese O objetivo deste trabalho foi descrever um caso fatal de salmonelose em uma fêmea primata não humana (Callithrix jacchus), originária de tráfico ilegal no Brasil. O sagui foi enviado para seção de quarentena do Zoológico Municipal de Guarulhos e morreu após um episódio de diarréia profusa. Achados d [...] e necropsia incluíram enterite mucosa, hepatomegalia e necrose do fígado. Fezes e fragmentos do fígado foram coletados para testes bacteriológicos e indicaram a presença de Salmonella sp.; subsequentemente caracterizada como sorotipo Yoruba. O perfil de suscetibilidade mostrou resistência somente à tetraciclina. A cepa foi positiva para os genes que codificam os fatores de virulência investigados (invA, sefC, pefA and spvC). Os resultados indicaram o risco de introdução de sorotipos patogênicos de Salmonella por primatas em cativeiro. Abstract in english The aim of this study was to describe a fatal salmonellosis case in a non-human female primate (Callithrix jacchus), found in the illegal pet trade in Brazil. The marmoset was sent to the quarantine section of the Guarulhos City Zoo and died in the sequence of an episode of profuse diarrhea. Necrops [...] y findings included mucous enteritis, and liver enlargement and necrosis. Feces and liver fragments were collected for bacteriological tests, which indicated the presence of Salmonella sp.; it was subsequently characterized as pertaining to the Yoruba serotype. The susceptibility profile demonstrated resistance to tetracycline only. The strain was positive for genes that encoded the virulence factors investigated (invA, sefC, pefA and spvC). The results indicated the risk of introduction of Salmonella pathogenic serotypes in primates in captivity.

Terezinha, Knöbl; Leliane T., Rocha; Márcia C., Menão; Cláudia A.S., Igayara; Renata, Paixão; Andréa M., Moreno.

80

Inhibition of polymerase chain reaction for the detection of Escherichia coli O157:H7 and Salmonella enterica on walnut kernels.  

Science.gov (United States)

The aim of this study was to determine whether Escherichia coli O157:H7 can be reliably detected and isolated from walnut kernels using standard methods of analysis. The limit of detection approached 1 cell per analytical unit (25 g) for E. coli O157:H7 on walnut kernels enriched in modified tryptic soy broth with 20 ?g/ml novobiocin and plating onto selective agar media. The presence of PCR inhibitors in walnut kernels was indicated by the failure to detect E. coli O157:H7 from culture positive enrichment broths analysed by PCR, with two separate polymerase and reagent compositions (Dupont BAX E. coli O157:H7 MP system, Promega GoTaq Green for stx) and three methods of template preparation (DuPont BAX, Qiagen DNeasy, Bio-Rad InstaGene). PCR inhibition was overcome by 1:100 dilution in TE buffer of the DNeasy or InstaGene template. PCR inhibition was not relieved by dilution of the BAX template. Similar results were observed for walnut kernels inoculated with Salmonella enterica and analysed for invA, indicating that PCR inhibition is not specific to the organism or primer/template. These results indicate that analysis of walnut kernels for pathogens should be with culture based methods or use protocols for DNA template preparation modified to remove or dilute inhibitors and the need for internal amplification controls in PCR methods. PMID:23628609

Ganz, Kyle; Gill, Alex

2013-08-01

 
 
 
 
81

Detección de Salmonella y coliformes fecales en agua de uso agrícola para la producción de melón "Cantaloupe" / Detection of Salmonella and fecal coliforms in water for agricultural use destined to melon"Cantaloupe"  

Scientific Electronic Library Online (English)

Full Text Available SciELO Mexico | Language: Spanish Abstract in spanish El agua que se utiliza en la producción de cultivos hortofrutícolas representa una fuente potencial de microorganismos que ocasionan enfermedades de transmisión alimentaria. Con el objetivo de evaluar la calidad sanitaria de diferentes fuentes de agua empleadas en la producción de melón Cantaloupe ( [...] Cucumis melo L. [grupo reticulatus] cv. Ovación y Caminos), en Zirándaro de los Chávez, Guerrero, se analizaron 71 muestras de agua provenientes de dos unidades de campo (23) y de una unidad de empaque (48) mediante métodos bacteriológicos convencionales para la detección deSalmonella spp, y por el método de filtración en membrana y el crecimiento en medios selectivos, para la detección de coliformes fecales, como indicadores de contaminación fecal. Del total de muestras de agua analizadas sólo tres muestras de campo resultaron positivas a la presencia de Salmonella spp. y nueve muestras, siete de campo y dos de la unidad de empaque, resultaron positivas a coliformes fecales. Salmonella spp. y coliformes fecales se detectaron principalmente en muestras de agua no clorada a 29 °C y 7.5 de pH, en promedio. En pruebas de reacción en cadena de la polimerasa (PCR), dos de cuatro cepas presuntivas de Salmonella ssp. dieron amplificaciones positivas con los iniciadores Sal-3 y Sal-4, y invA-1 e invA-2; de las otras dos, sólo dieron amplificación positiva con Sal-3 y Sal-4. Aparentemente se tiene más de una raza o serovar de Salmonella en la región. Estos resultados sugieren que algunas de las fuentes de agua empleadas en la producción de melón Cantaloupe en Zirándaro de los Chávez, Guerrero, no cumplen con la normatividad sanitaria por lo que estas fuentes deben establecerse como puntos prerequisitos de control para evitar la contaminación de melones frescos con patógenos de humanos. Abstract in english Water used in the production of horticultural crops represents a potential source of microorganisms that cause food-transmitted diseases. In order to evaluate the sanitary quality of different agricultural water sources used in the production of Cantaloupe melon (Cucumis melo L. [group reticulatus] [...] cv. Ovacion and Caminos), in Zirandaro Chavez, Guerrero, 71 water samples were analyzed from two field units (23) and one packaging house unit (48) through traditional bacteriological methods, to detect Salmonella spp, and the filtering membrane method and growth selective media, to detect fecal coliforms, as fecal contamination indicators. Of the total analyzed water samples only three field samples were positive to Salmonella spp. and nine samples, seven coming from the field and two from the packaging house, were positive to fecal coliforms. Salmonella spp. and fecal coliforms were detected mainly in non-chlorinated water samples at 29 °C and pH of 7.5, on average. Two out of four presumptive Salmonella spp. isolates were confirmed by the polymerase chain reaction (PCR) using primers Sal-3 and Sal-4, and invA-1 and invA-2; the other two were only confirmed with Sal-3 y Sal-4. Apparently more than one race or serovar of Salmonella spp. are present in this region. Results suggest that some water sources used in the Cantaloupe melon production in Zirandaro Chavez, Guerrero, do not meet sanitary standards therefore these sources should be considered as critical control points to prevent fresh melon contamination with human pathogens.

Carmela, Hernández-Domínguez; Ana María, Hernández-Anguiano; Cristóbal, Cháidez-Quiroz; Gilberto, Rendón-Sánchez; Trevor, Suslow.

2008-03-01

82

Detección de Salmonella y coliformes fecales en agua de uso agrícola para la producción de melón "Cantaloupe" / Detection of Salmonella and fecal coliforms in water for agricultural use destined to melon"Cantaloupe"  

Scientific Electronic Library Online (English)

Full Text Available SciELO Mexico | Language: Spanish Abstract in spanish El agua que se utiliza en la producción de cultivos hortofrutícolas representa una fuente potencial de microorganismos que ocasionan enfermedades de transmisión alimentaria. Con el objetivo de evaluar la calidad sanitaria de diferentes fuentes de agua empleadas en la producción de melón Cantaloupe ( [...] Cucumis melo L. [grupo reticulatus] cv. Ovación y Caminos), en Zirándaro de los Chávez, Guerrero, se analizaron 71 muestras de agua provenientes de dos unidades de campo (23) y de una unidad de empaque (48) mediante métodos bacteriológicos convencionales para la detección deSalmonella spp, y por el método de filtración en membrana y el crecimiento en medios selectivos, para la detección de coliformes fecales, como indicadores de contaminación fecal. Del total de muestras de agua analizadas sólo tres muestras de campo resultaron positivas a la presencia de Salmonella spp. y nueve muestras, siete de campo y dos de la unidad de empaque, resultaron positivas a coliformes fecales. Salmonella spp. y coliformes fecales se detectaron principalmente en muestras de agua no clorada a 29 °C y 7.5 de pH, en promedio. En pruebas de reacción en cadena de la polimerasa (PCR), dos de cuatro cepas presuntivas de Salmonella ssp. dieron amplificaciones positivas con los iniciadores Sal-3 y Sal-4, y invA-1 e invA-2; de las otras dos, sólo dieron amplificación positiva con Sal-3 y Sal-4. Aparentemente se tiene más de una raza o serovar de Salmonella en la región. Estos resultados sugieren que algunas de las fuentes de agua empleadas en la producción de melón Cantaloupe en Zirándaro de los Chávez, Guerrero, no cumplen con la normatividad sanitaria por lo que estas fuentes deben establecerse como puntos prerequisitos de control para evitar la contaminación de melones frescos con patógenos de humanos. Abstract in english Water used in the production of horticultural crops represents a potential source of microorganisms that cause food-transmitted diseases. In order to evaluate the sanitary quality of different agricultural water sources used in the production of Cantaloupe melon (Cucumis melo L. [group reticulatus] [...] cv. Ovacion and Caminos), in Zirandaro Chavez, Guerrero, 71 water samples were analyzed from two field units (23) and one packaging house unit (48) through traditional bacteriological methods, to detect Salmonella spp, and the filtering membrane method and growth selective media, to detect fecal coliforms, as fecal contamination indicators. Of the total analyzed water samples only three field samples were positive to Salmonella spp. and nine samples, seven coming from the field and two from the packaging house, were positive to fecal coliforms. Salmonella spp. and fecal coliforms were detected mainly in non-chlorinated water samples at 29 °C and pH of 7.5, on average. Two out of four presumptive Salmonella spp. isolates were confirmed by the polymerase chain reaction (PCR) using primers Sal-3 and Sal-4, and invA-1 and invA-2; the other two were only confirmed with Sal-3 y Sal-4. Apparently more than one race or serovar of Salmonella spp. are present in this region. Results suggest that some water sources used in the Cantaloupe melon production in Zirandaro Chavez, Guerrero, do not meet sanitary standards therefore these sources should be considered as critical control points to prevent fresh melon contamination with human pathogens.

Carmela, Hernández-Domínguez; Ana María, Hernández-Anguiano; Cristóbal, Cháidez-Quiroz; Gilberto, Rendón-Sánchez; Trevor, Suslow.

83

Salmonella Susceptibility  

Science.gov (United States)

Nontyphoidal Salmonella (NTS) are a group of enteric bacteria can lead to life-threatening bacteremia in those with weakened immune systems. MacLennan et al. identify an immune response that may have important implications for the development of a vaccine against NTS.

Susan Moir (National Institutes of Health;); Anthony Fauci (National Institutes of Health;)

2010-04-23

84

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

Science.gov (United States)

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. PMID:25273985

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

2014-01-01

85

Comparison of culture methods for isolation of salmonella in yak fecal samples.  

Science.gov (United States)

To compare the effectiveness of culture methods for identifying yak Salmonella, three selective enrichment broths (SC, TTB, MSRV) and three media (SS, XLD, CAS) for detecting Salmonella were evaluated in this study. The results showed that TTB broth was better than SC broths and MSRV broths, and SS medium has the highest isolation rate, significantly higher than those of CAS and XLD media (P < 0.05). It is worth noticing that there was no overlapping of the positive results given by TTB, SC and MSRV broths. In addition, all of the yak Salmonella isolates were detected positive by the five reported PCR assays, targeting the invA, srfC, invE, stn and 16S-23S rRNA genes. The combination of TTB and MSRV broths and SS and CAS media (or XLD) recommended in this study was relatively efficient in recovering Salmonella from yak feces, and the five PCR assays can be successfully used to identify yak Salmonella. PMID:25320426

Yue, Hua; Zhang, Bin; Zhu, Xiaoxia; Zhang, Huanrong; Tang, Cheng

2014-06-01

86

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

Science.gov (United States)

Typhoid fever is a public health problem, especially among young children in developing countries. To address this need, a glycoconjugate vaccine Vi-CRM???, composed of the polysaccharide antigen Vi covalently conjugated to the non-toxic mutant of diphtheria toxin CRM???, is under development. Here, we assessed the antibody and cellular responses, both local and systemic, following subcutaneous injection of Vi-CRM???. 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-CRM??? was observed in spleen and mesenteric lymph nodes. These data confirm the immunogenicity of Vi-CRM??? and demonstrate that the vaccine-specific antibody and cellular immune responses are present also in the intestinal tract, thus strengthening the suitability of Vi-CRM??? as a promising candidate vaccine against Salmonella Typhi. PMID:22705173

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

2012-09-21

87

Salmonella: Annual Summary, 2005.  

Science.gov (United States)

This Annual Summary of the National Salmonella Surveillance System contains surveillance data on reported laboratory-confirmed Salmonella isolates in the United States for the year 2005. The National Salmonella Surveillance System collects reports of isol...

2005-01-01

88

Salmonella from Pocket Pets  

Science.gov (United States)

... Transplant Patients Infants and Young Children Publications & Materials Salmonella from Pocket Pets Share Compartir The Salmonella germ ... has been cleaned and disinfected. Tips for Preventing Salmonella from Rodents General Information on Washing your Hands ...

89

78 FR 42526 - Salmonella  

Science.gov (United States)

...Docket No. FDA-2013-D-0254] Salmonella Contamination of Dry Dog Food; Withdrawal...CPG) entitled ``Sec. 690.700 Salmonella Contamination of Dry Dog Food...the CGP entitled ``Sec. 690.700 Salmonella Contamination of Dry Dog Food...

2013-07-16

90

A rapid and direct real time PCR-based method for identification of Salmonella spp.  

DEFF Research Database (Denmark)

The aim of this work was the validation of a rapid, real-time PCR assay based on TaqMan((R)) technology for the unequivocal identification of Salmonella spp. to be used directly on an agar-grown colony. A real-time PCR system targeting at the Salmonella spp. invA gene was optimized and validated through a four times repeated blind experiment performed in two different laboratories including 50 Salmonella spp. with representative strains from each of the 5 different Salmonella subgenera and 30 non-Salmonella strains. Both parameters DeltaR(n) (fluorescence intensity of template through a normalized reporter value) and C-T (cycle at which the fluorescence intensity achieved a pre-established threshold) were analyzed. Overall mean DeltaR(n) and C-T values for Salmonella strains (2.14 +/- 0.87 and 15.30 +/- 0.90, respectively) were statistically different from values for non-Salmonella strains, allowing the establishment of cut-off DeltaR(n) and C-T values based on 95% confidence intervals that allowed the correct identification of all strains tested in each independent experiment. The accuracy of this assay in terms of inclusivity and exclusivity was 100%. Moreover, the PCR system proved to be especially convenient because the pre-mix containing all PCR reagents except for the bacterial cells could be kept at -20 degreesC for at least I month before its use. The optimized TaqMan((R)) real-time PCR assay is a useful, simple and rapid method for routine identification of Salmonella spp., irrespective of the particular subgenus.

Hoorfar, Jeffrey

2003-01-01

91

PREVALENCE OF DRUG RESISTANCE AND VIRULENCE FEATURES IN Salmonella spp. ISOLATED FROM FOODS ASSOCIATED OR NOT WITH SALMONELLOSIS IN BRAZIL.  

Science.gov (United States)

Salmonella is the most common etiological agent of cases and outbreaks of foodborne diarrheal illnesses. The emergence and spread of Salmonella spp., which has become multi-drug resistant and potentially more pathogenic, have increased the concern with this pathogen. In this study, 237 Salmonella spp., associated or not with foodborne salmonellosis in Brazil, belonging mainly to serotype Enteritidis, were tested for antimicrobial susceptibility and the presence of the virulence genes spvC, invA, sefA and pefA. Of the isolates, 46.8% were sensitive to all antimicrobials and 51.9% were resistant to at least one antimicrobial agent. Resistance to more than one antimicrobial agent was observed in 10.5% of the strains. The highest rates of resistance were observed for streptomycin (35.9%) and nalidixic acid (16.9%). No strain was resistant to cefoxitin, cephalothin, cefotaxime, amikacin, ciprofloxacin and imipenem. The invA gene was detected in all strains. Genes spvC and pefA were found in 48.1% and 44.3% of strains, respectively. The gene sefA was detected in 31.6% of the strains and only among S. Enteritidis. Resistance and virulence determinants were detected in Salmonella strains belonging to several serotypes. The high rates of antibiotic-resistance in strains isolated from poultry products demonstrate the potential risk associated with the consumption of these products and the need to ensure good food hygiene practices from farm to table to reduce the spread of pathogens relevant to public health. PMID:25351537

Rowlands, Ruth Estela Gravato; Ristori, Christiane Asturiano; Ikuno, Alice A; Barbosa, Maria Luisa; Jakabi, Miyoko; Franco, Bernadette Dora Gombossy de Melo

2014-12-01

92

PERFIL SOROLÓGICO E DE ISOLAMENTO DE Salmonella sp. EM SUÍNOS NO INÍCIO DA TERMINAÇÃO E AO ABATE SEROLOGY AND ISOLATION OF SALMONELLA SP. IN PIGS AT THE FINISHING SITE AND AT SLAUGHTER  

Directory of Open Access Journals (Sweden)

Full Text Available Estudos que elucidem a cadeia de transmissão de Salmonella enterica nos sistemas de produção de suínos são importantes para que seja possível implementar programas de controle da infecção. O objetivo deste estudo foi comparar o índice de animais positivos para Salmonella sp. no início da fase de terminação e ao abate e identificar possíveis fontes de contaminação no período. Em três granjas terminadoras, coletaram-se: suabes de superfície nas baias e nos silos durante o vazio sanitário; amostras de fezes e sangue dos animais no dia do alojamento; alíquotas de todos os lotes de ração e amostras de sangue, linfonodos mesentéricos (LM e conteúdo intestinal (CI ao abate. As amostras de sangue foram submetidas a teste de ELISA-LPS para Salmonella Typhimurium. Nas demais amostras, pesquisou-se a presença de Salmonella sp. As amostras de ração foram adicionalmente submetidas à técnica da Reação em Cadeia da Polimerase (PCR amplificando o gene invA. Todos os animais foram negativos para presença de Salmonella sp. nas fezes no início da terminação; entretanto, em duas granjas havia animais soropositivos (12% e 28%, respectivamente. Em duas granjas havia contaminação residual no ambiente e na terceira granja, em um dos lotes de ração, detectou-se a presença de Salmonella sp. pela PCR. Ao abate, acima de 90% dos animais foram positivos no teste de ELISA-LPS, sendo que em todos os lotes encontrou-se um número variável (12-92% de portadores em LM e CI. A partir disso, concluiu-se que a terminação foi a fase crítica para a amplificação da infecção por Salmonella sp., sendo a presença residual do microrganismo na granja e o fornecimento de ração contaminada fontes prováveis de infecção.

PALAVRAS-CHAVES: Abate, isolamento, Salmonella, sorologia, suíno, terminação.
Studies assessing the Salmonella transmission chain in pig herds are the first step to start a control program.  The aims of this study were to compare the prevalence of Salmonella positive pigs at the beginning of the finishing phase and at slaughter, and to identify the possible sources of contamination in the farms. In three finishing farms, environmental swabs from the barns and from the feed silos were collected during the sanitary emptiness. Furthermore, samples of feces and blood from the animals on the day of housing; and aliquots from all feed lots were taken. At slaughter, blood, mesenteric lymph nodes (LM and intestinal content (CI were sampled. Blood samples were submitted to a S. Typhimurium ELISA-LPS test. All other samples were submitted to a Salmonella isolation protocol. Feed samples were also submitted to Polymerase Chain Reaction (PCR targeting the invA gene. Feces samples from all pigs were Salmonella negative at the beginning of the finishing phase, in two farms seropositive animals were found. In two farms, residual environmental contamination was detected, and, in the third farm, one of the feed batches was Salmonella positive on the PCR assay. At slaughter, over 90% of the animals were positive on the ELISA-LPS test and, in all cohorts, a variable number (12%-92% of carriers in LM and CI was detected. From this on, it was concluded that the finishing phase was critical for the amplification of Salmonella infection, and the residual environmental contamination in the farms as well as Salmonella positive feed batches were the probable infection sources.

KEY WORDS: Isolation, finishing and slaughter, Salmonella, serology, swine.

Monika Müller

2009-09-01

93

Immediate differentiation of salmonella-resembling colonies on brilliant green agar  

DEFF Research Database (Denmark)

A rapid biochemical system (OBIS) based on immediate enzymatic differentiation of Citrobacter, Proteus, Providencia, Hafnia and Morganella spp. from Salmonella on brilliant green agar was evaluated A total of 96 field isolates from various Salmonella serotypes, 18 Citrobacter freundii and 25 isolates of other Enterobacteriaceae were tested All Salmonella isolates were identified correctly by the kit, and none of the Enterobacteriaceae isolates were identified as Salmonella. The results indicate complete specificity for Salmonella colonies on brilliant green agar.

Jensen, Annette Nygaard; Hoorfar, Jeffrey

2000-01-01

94

Efficient mobilization of a resistance derivative of pSLT, the virulence plasmid specific of Salmonella enterica serovar Typhimurium, by an IncI1 plasmid.  

Science.gov (United States)

pUO-StVR2 is a derivative of pSLT, the virulence plasmid specific of Salmonella enterica serovar Typhimurium, which confers multidrug resistance. This plasmid is widespread among closely related isolates of S. Typhimurium, and often coexists with other plasmids like pStR12. The latter belongs to incompatibility group IncI1, was assigned to ST48 by pMLST (plasmid multilocus sequence typing), and confers resistance to streptomycin/spectinomycin, chloramphenicol, trimethoprim and sulphonamides, with the responsible genes (aadA1/aadA2, cmlA1, dfrA12 and sul3) located on a sul3-class 1 integron. When using clinical isolates of S. Typhimurium containing one (pUO-StVR2) or both (pUO-StVR2 and pStR12) plasmids as donors and Escherichia coli K12 J53 resistant to rifampicin as recipient, the conjugation frequencies of pUO-StVR2 and pStR12 were 10?? and 10?³-10?? transconjugants/donor, respectively, while the transfer frequency of pUO-StVR2 increased 10² up to 10? times through mobilization by pStR12, depending on the donor strain and experimental conditions. Mobilization of pUO-StVR2, a plasmid which encodes virulence and resistance functions, by compatible plasmids which coexist in the same bacterium can facilitate the spread of these properties in S. Typhimurium, one of the most common serovars of S. enterica. PMID:23541844

Montero, Ignacio; Herrero-Fresno, Ana; Rodicio, Rosaura; Rodicio, M Rosario

2013-07-01

95

Mutagenicity and antimutagenicity testing of six chemicals associated with the pungent properties of specific spices as revealed by the Ames Salmonella/microsomal assay.  

Science.gov (United States)

Three compounds, capsaicin, thymol and borneol, were initially screened for mutagenic activity using Salmonella typhimurium strains TA97, TA98 and TA100, with and without S9 metabolic activation, and 20 min standard preincubation time. Three other compounds, allyl isothiocyanate, eugenol and cinnamaldehyde, were screened for mutagenic activity as above, but with a prolonged, nonstandard preincubation time of up to 120 minutes. All six test compounds used in the assays are associated with the pungent properties of some specific spices in which the test compounds can be found to exist naturally. The first objective of this study was to observe if mutagenic activity can be correlated to the pungent properties of these six test compounds. However, due to toxicity and the observation that only capsaicin was mutagenic, using strain TA100 in the presence of S9 metabolic activation, it was not possible to deduce any relationship between mutagenicity and the test chemicals' pungent properties. Naturally occurring capsaicin, found in the spice Capsicum annum, was detected and quantified using thin layer and gas chromatographic techniques. The final objective was to detect the presence of antimutagenic factor(s) in C. annum that would suppress the mutagenicity of capsaicin. When the mutagenic capsaicin and 2-aminoanthracene were assayed in the presence of C. annum acetone extract, using strain TA100 with S9 metabolic activation, the mutagenic response of both the mutagens were reduced by approximately 50%. Assaying capsaicin and 2-aminoanthracene in the presence of chlorophyll, the mutagenic response of the two mutagens was reduced by less than 40%. From this observation it was inferred that chlorophyll can successfully suppress the mutagenicity activities of capsaicin and 2-aminoanthracene, together with other antimutagenic factors that were present in the acetone extract of C. annum. PMID:7710293

Azizan, A; Blevins, R D

1995-02-01

96

Simple and rapid detection of Salmonella by direct PCR amplification of gene fimW.  

Science.gov (United States)

This study established a simple method of specifically detecting Salmonella species by amplifying fimW gene, which was involved in regulating Salmonella type I fimbriae expression. A pair of primers was designed to target and discriminate the 68 Salmonella strains of 23 Salmonella serovars available to us from 12 non-Salmonella strains of five different kinds of bacteria by polymerase chain reaction (PCR) amplification. Results showed that specific DNA fragment with an expected size of 477 bp was successfully amplified from all Salmonella serovars, while no target band was detected in non-Salmonella species. The sensitivity of this PCR-amplifying system reached to 1 pg DNA chromosome and 10(2) cfu of Salmonella enteritis strain CMCC(B) 50336. The above results demonstrated the method as a simple, sensitive, and specific way for Salmonella detection. PMID:24838665

Zhang, Jiang-ying; Dong, Li-wei; Ren, Qian; Wang, Xiao-zhou; Yang, Yi; Zhou, Wen; Zhu, Chun-hong; Meng, Xia; Zhu, Guo-qiang

2014-10-01

97

Genes de virulência e diversidade genética em Salmonella spp. isoladas de amostras de origem suína / Virulence genes and genetic diversity in Salmonella spp. isolated from samples of swine origin  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese A diversificação da produção industrial de alimentos de origem suína e o intercâmbio comercial de animais e seus derivados destinados ao consumo humano podem ser importantes disseminadores de sorovares de Salmonella spp. na cadeia alimentar. Objetivou-se avaliar em 86 cepas de Salmonella spp., isola [...] das em granja de terminação e no abate de suínos, a ocorrência de três genes de virulência (invA, agfA e lpfA), bem como a similaridade genética entre elas. A ocorrência do gene invA foi verificada em 100% das amostras. O gene lpfA foi detectado em 80,23% (69/86) das cepas, não foi detectado em S. Panama e estava presente em todas as cepas de S. Infantis. O gene agfA foi detectado em 63,95% (55/86) das amostras. S. Agona apresentou positividade para todos os genes de virulência estudados. A análise de homologia entre as cepas agrupou os diferentes sorovares em clusters. A similaridade foi independente do local de isolamento, o que demonstra a presença de clones ao longo da cadeia de produção e a existência de multiplicidade de fontes para a infecção dos animais, como a ração, e a contaminação cruzada das carcaças. A pesquisa de genes de virulência e a avaliação da proximidade gênica permitem a caracterização e um maior entendimento sobre cepas de Salmonella circulantes na cadeia produtiva de suínos e, assim, podem subsidiar medidas de controle durante o processo produtivo com o objetivo de garantir a saúde do consumidor. Abstract in english The diversification of industrial food production of swine origin and trade of animals and their derivatives for human consumption may be important disseminators of serovars of Salmonella spp. in the food chain. This study aimed to evaluate 86 strains of Salmonella spp. isolated form in the finishin [...] g and slaughter of pigs, the occurrence of three virulence genes (invA, agfa and lpfA), as well as the genetic similarity between them. The occurrence of gene invA was observed in 100% of the samples. The gene lpfA was detected in 80.23% (69/86) strains and is not detected in S. Panama, but present in all strains of S. Infantis. The gene agfA was detected in 63.95% (55/86). S. Agona was positive for all virulence genes studied. The analysis of homology between the different serovars grouped the isolates in clusters. The similarity was regardless of the location of isolation, demonstrating the presence of clones along the production chain and that there are multiple sources for the infection of animals, such as feed, and cross-contamination of carcasses. A survey of virulence genes and evaluation of gene proximity allow characterization and better understanding of Salmonella strains circulating in the pig production chain, thus being able to support control measures during the production process in order to ensure consumer health.

M.S., Moura; R.P., Oliveira; R.T., Melo; E.P., Mendonça; B.B., Fonseca; D.A., Rossi.

1367-13-01

98

Excreción fecal de Salmonella Albany, su aislamiento en la ración alimenticia y repercusión en el estado de salud de un ocelote (Leopardus pardalis en cautiverio  

Directory of Open Access Journals (Sweden)

Full Text Available Los serotipos de Salmonella especie enterica son los responsables del 99% de las salmonelosis en humanos y animales, en particular, Salmonella enterica serovariedad Albany se ha identificado en canales de pollo, por lo que representa un riesgo para la salud humana y animal. Se aisló Salmonella enterica serovariedad Albany a partir de heces de un ocelote macho (Leopardus pardalis, cautivo en el zoológico de Culiacán, Sinaloa, México, y de pollo crudo (alimento del felino. El patrón por electroforesis en campo pulsado (PFGE con la enzima Xba I fue idéntico en ambos aislados, lo que indica que la fuente de infección fue el pollo crudo. Cinco meses después de haber aislado las bacterias de las heces, se realizó estudio post mortem del felino anteriormente mencionado, y se observó macroscópicamente: enterocolitis hemorrágica severa y fibrosis renal; y microscópicamente: necrosis de vellosidades y de criptas e infiltrado mononuclear linfocitario severo en íleon, además nefritis intersticial severa multifocal y fibrosis en riñón. A partir de muestras intestinales se amplificó el gen invA que confirma la infección por Salmonella. Los diagnósticos microbiológico, molecular e histopatológico sugieren que la muerte del felino se debió a la infección causada por la ingesta de pollo crudo contaminado con Salmonella enterica serovariedad Albany. Este caso clínico confirma la importancia que tienen los animales que excretan Salmonella vía fecal y describe la relación epidemiológica-molecular de los aislamientos obtenidos de heces y alimento, lo que permitió esclarecer la fuente primaria de infección.

Gabriela Silva-Hidalgo

2012-01-01

99

Salmonella Infections (For Parents)  

Science.gov (United States)

... pets, particularly if you have very young children. Hand washing is a powerful way to guard against salmonella ... is often the source of salmonella contamination, so hand washing is extremely important, particularly after using the toilet ...

100

Tentative Colistin Epidemiological Cut-Off Value for Salmonella spp.  

DEFF Research Database (Denmark)

The objective of this research was to determine minimal inhibitory concentration (MIC) population distributions for colistin for Salmonella on subtype level. Furthermore, we wanted to determine if differences in MIC for colistin could be explained by mutations in pmrA or pmrB encoding proteins involved in processes that influence the binding of colistin to the cell membrane. During 2008–2011, 6,583 Salmonella enterica subsp. enterica isolates of human origin and 1931 isolates of animal/meat origin were collected. The isolates were serotyped, and susceptibility was tested towards colistin (range 1–16 mg/L). Moreover, 37 isolates were tested for mutations in pmrA and pmrB by polymerase chain reaction (PCR) and DNA sequencing. MIC distribution for colistin at serotype level showed that Salmonella Dublin (n=198) followed by Salmonella Enteritidis (n=1247) were less susceptible than “other” Salmonella serotypes originating from humans (n=5,274) and Salmonella Typhimurium of animal/meat origin (n=1794). MIC was ?1 mg/L for 98.9% of “other” Salmonella serotypes originating from humans, 99.4% of Salmonella Typhimurium, 61.3% of Salmonella Enteritidis, and 12.1% of Salmonella Dublin isolates. Interestingly, Salmonella Dublin and Salmonella Enteritidis belong to the same O-group (O:1, 9,12), suggesting that surface lipopolysaccharides (LPS) of the cell (O-antigen) play a role in colistin susceptibility. The epidemiological cut-off value of >2 mg/L for colistin suggested by European Committee on Antimicrobial Susceptibility Testing (EUCAST) is placed inside the distribution for both Salmonella Dublin and Salmonella Enteritidis. All tested Salmonella Dublin isolates, regardless of MIC colistin value, had identical pmrA and pmrB sequences. Missense mutations were found only in pmrA in one Salmonella Reading and in pmrB in one Salmonella Concord isolate, both with MIC of ?1 for colistin. In conclusion, our study indicates that missense mutations are not necessarily involved in increased MICs for colistin. Increased MICs for colistin seemed to be linked to specific serotypes (Salmonella Dublin and Salmonella Enteritidis). We recommend that Salmonella with MIC of >2 mg/L for colistin be evaluated on the serovar level.

AgersØ, Yvonne; Torpdahl, Mia

2012-01-01

 
 
 
 
101

Gene-specific effects of antisense phosphorodiamidate morpholino oligomer-peptide conjugates on Escherichia coli and Salmonella enterica serovar typhimurium in pure culture and in tissue culture.  

Science.gov (United States)

The objective was to improve efficacy of antisense phosphorodiamidate morpholino oligomers (PMOs) by improving their uptake into bacterial cells. Four different bacterium-permeating peptides, RFFRFFRFFXB, RTRTRFLRRTXB, RXXRXXRXXB, and KFFKFFKFFKXB (X is 6-aminohexanoic acid and B is beta-alanine), were separately coupled to two different PMOs that are complementary to regions near the start codons of a luciferase reporter gene (luc) and a gene required for viability (acpP). Luc peptide-PMOs targeted to luc inhibited luciferase activity 23 to 80% in growing cultures of Escherichia coli. In cell-free translation reactions, Luc RTRTRFLRRTXB-PMO inhibited luciferase synthesis significantly more than the other Luc peptide-PMOs or the Luc PMO not coupled to peptide. AcpP peptide-PMOs targeted to acpP inhibited growth of E. coli or Salmonella enterica serovar Typhimurium to various extents, depending on the strain. The concentrations of AcpP RFFRFFRFFXB-PMO, AcpP RTRTRFLRRTXB-PMO, AcpP KFFKFFKFFKXB-PMO, and ampicillin that reduced CFU/ml by 50% after 8 h of growth (50% inhibitory concentration [IC(50)]) were 3.6, 10.8, 9.5, and 7.5 microM, respectively, in E. coli W3110. Sequence-specific effects of AcpP peptide-PMOs were shown by rescuing growth of a merodiploid strain that expressed acpP with silent mutations in the region targeted by AcpP peptide-PMO. In Caco-2 cultures infected with enteropathogenic E. coli (EPEC), 10 microM AcpP RTRTRFLRRTXB-PMO or AcpP RFFRFFRFFXB-PMO essentially cleared the infection. The IC(50) of either AcpP RTRTRFLRRTXB-PMO or AcpP RFFRFFRFFXB-PMO in EPEC-infected Caco-2 culture was 3 microM. In summary, RFFRFFRFFXB, RTRTRFLRRTXB, or KFFKFFKFFXB, when covalently bonded to PMO, significantly increased inhibition of expression of targeted genes compared to PMOs without attached peptide. PMID:16870773

Tilley, Lucas D; Hine, Orion S; Kellogg, Jill A; Hassinger, Jed N; Weller, Dwight D; Iversen, Patrick L; Geller, Bruce L

2006-08-01

102

A carbon nanotube immunosensor for Salmonella  

Science.gov (United States)

Antibody-functionalized carbon nanotube devices have been suggested for use as bacterial detectors for monitoring of food purity in transit from the farm to the kitchen. Here we report progress towards that goal by demonstrating specific detection of Salmonella in complex nutrient broth solutions using nanotube transistors functionalized with covalently-bound anti-Salmonella antibodies. The small size of the active device region makes them compatible with integration in large-scale arrays. We find that the on-state current of the transistor is sensitive specifically to the Salmonella concentration and saturates at low concentration (Salmonella and other bacteria types, with no sign of saturation even at much larger concentrations (108 cfu/ml).

Lerner, Mitchell B.; Goldsmith, Brett R.; McMillon, Ronald; Dailey, Jennifer; Pillai, Shreekumar; Singh, Shree R.; Johnson, A. T. Charlie

2011-12-01

103

Development of a real-time PCR melt curve assay for simultaneous detection of virulent and antibiotic resistant Salmonella.  

Science.gov (United States)

Multiple drug resistance in Salmonella is an emerging problem in the area of food safety. Depending on the virulence and antibiotic resistance characteristics of the Salmonella strain, infections of varying severity could result. In this study, a multiplex melt curve real-time PCR assay for the detection of virulent and antibiotic resistance strains of Salmonella was developed with two primer sets. The first set targets the virulence gene, invasin (invA), and tetracycline (tetG), streptomycin (aadA2) and sulphonamide (sulI) antibiotic resistance genes, and the second set amplifies ampicillin (blaPSE,blaTEM) and chloramphenicol (floR) resistance genes. The multiplex assay was evaluated using 41 Salmonella strains and was further tested on eight different artificially inoculated food samples. The fluorescent DNA intercalating dye, SYTO9, generated high resolution melt curve peaks and, hence, was used for the development of the assay. This multiplex assay worked efficiently over a DNA concentration range of 20 ng-200 fg and showed a sensitivity of 290 CFU/mL with serially diluted broth cultures. The detection limit for un-enriched artificially inoculated food samples was 10(4) CFU/g, but an enrichment period of 6 h allowed for detection of 10 CFU/g of cells in the samples. PMID:25084639

Singh, Prashant; Mustapha, Azlin

2014-12-01

104

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese INTRODUÇÃO: A diarreia infecciosa aguda é considerada um problema de saúde pública associado a uma ampla gama de agentes etiológicos, entre os quais destaca-se Salmonella enterica, causa importante de diarreia inflamatória em países desenvolvidos e em desenvolvimento. OBJETIVOS: Avaliar a distribuiç [...] ão de S. enterica em crianças com diarreia aguda em Belo Horizonte e caracterizar as amostras isoladas. MATERIAL E MÉTODO: O grupo de estudo consistiu de 157 crianças de nível socioeconômico baixo. Espécimes fecais foram empregados para pesquisa de leucóc itos e cultivo de Salmonella. As amostras isoladas foram sorotipadas e submetidas à avaliação do perfil de suscetibilidade a antimicrobianos, da produção de betalactamases de amplo espectro (ESBL) e da presença de marcadores de virulência (invA, iroB e spvC). RESULTADOS: Cinco/3,2% crianças apresentaram-se infectadas por S. enterica; três/60%, por S. enterica Typhimurium; uma/20%, por S. enterica Enteritidis; e uma/20%, por S. enterica subsp. enterica sorotipo 8,20:z4,z23:-. Leucócitos fecais foram detectados em dois dos cinco espécimes positivos para S. enterica. As amostras isoladas de três crianças apresentaram resistência a ácido nalidíxico, ácido nalidíxico + cloranfenicol e ácido nalidíxico + cloranfenicol + ampicilina. Nenhuma amostra produziu ESBL. Todas as amostras albergavam os genes invA e iroB. O marcador spvC foi observado em amostras isoladas de duas crianças infectadas por S. Typhimurium e S. Enteritidis. CONCLUSÃO: Os resultados demonstram que diarreia associada a S. enterica é raramente observada entre crianças da nossa região e indicam a necessidade de avaliação periódica do perfil de suscetibilidade a antimicrobianos da bactéria para orientar o estabelecimento de antibioticoterapia, quando indicada. Abstract in english Introduction: Acute infectious diarrhea is still regarded as a public health problem associated with a wide range of etiologic agents, from which Salmonella enterica is particularly worth mentioning inasmuch as it is a major cause of inflammatory diarrhea in both developed and developing countries. [...] Objective: To assess the distribution of S. enterica among children with acute diarrhea in Belo Horizonte and to characterize bacterium isolates. Material and methods: The study group comprised a total of 157 children from low socioeconomic background. Stool samples were collected for leukocyte analysis and Salmonella bacterial culture. The isolates were serotyped and evaluated as to antimicrobial susceptibility profile, extended-spectrum ?-lactamases (ESBL) production, and presence of virulence markers (invA, iroB, and spvC). RESULTS: A total of 5/3.2% children were infected by S. enterica, 3/60% by S. enterica Typhimurium, 1/20% by S. enterica Enteritidis and 1/20% S. enterica subsp. enterica serotype 8.20:z4,z23:-. Fecal leucocytes were detected in two out of five fecal specimens positive for S. enterica. Isolates from three children were resistant to nalidixic acid, nalidixic acid + chloramphenicol, and nalidixic acid + chloramphenicol + ampicillin. ESBL production was not detected. All samples presented invA and iroB genes. spvC marker was observed in isolates from two children infected by S. Typhimurium and S. Enteritidis. Conclusion: The results demonstrate that S. enterica infection is uncommon among children from our region. Furthermore, they indicate the need for periodic monitoring of bacterial antimicrobial susceptibility profile in order to establish suitable antimicrobial therapy when required.

Mireille Ângela Bernardes, Sousa; Edilberto Nogueira, Mendes; Francisco José, Penna; Luciano Amedée, Péret-Filho; Paula Prazeres, Magalhães.

2013-02-01

105

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese INTRODUÇÃO: A diarreia infecciosa aguda é considerada um problema de saúde pública associado a uma ampla gama de agentes etiológicos, entre os quais destaca-se Salmonella enterica, causa importante de diarreia inflamatória em países desenvolvidos e em desenvolvimento. OBJETIVOS: Avaliar a distribuiç [...] ão de S. enterica em crianças com diarreia aguda em Belo Horizonte e caracterizar as amostras isoladas. MATERIAL E MÉTODO: O grupo de estudo consistiu de 157 crianças de nível socioeconômico baixo. Espécimes fecais foram empregados para pesquisa de leucóc itos e cultivo de Salmonella. As amostras isoladas foram sorotipadas e submetidas à avaliação do perfil de suscetibilidade a antimicrobianos, da produção de betalactamases de amplo espectro (ESBL) e da presença de marcadores de virulência (invA, iroB e spvC). RESULTADOS: Cinco/3,2% crianças apresentaram-se infectadas por S. enterica; três/60%, por S. enterica Typhimurium; uma/20%, por S. enterica Enteritidis; e uma/20%, por S. enterica subsp. enterica sorotipo 8,20:z4,z23:-. Leucócitos fecais foram detectados em dois dos cinco espécimes positivos para S. enterica. As amostras isoladas de três crianças apresentaram resistência a ácido nalidíxico, ácido nalidíxico + cloranfenicol e ácido nalidíxico + cloranfenicol + ampicilina. Nenhuma amostra produziu ESBL. Todas as amostras albergavam os genes invA e iroB. O marcador spvC foi observado em amostras isoladas de duas crianças infectadas por S. Typhimurium e S. Enteritidis. CONCLUSÃO: Os resultados demonstram que diarreia associada a S. enterica é raramente observada entre crianças da nossa região e indicam a necessidade de avaliação periódica do perfil de suscetibilidade a antimicrobianos da bactéria para orientar o estabelecimento de antibioticoterapia, quando indicada. Abstract in english Introduction: Acute infectious diarrhea is still regarded as a public health problem associated with a wide range of etiologic agents, from which Salmonella enterica is particularly worth mentioning inasmuch as it is a major cause of inflammatory diarrhea in both developed and developing countries. [...] Objective: To assess the distribution of S. enterica among children with acute diarrhea in Belo Horizonte and to characterize bacterium isolates. Material and methods: The study group comprised a total of 157 children from low socioeconomic background. Stool samples were collected for leukocyte analysis and Salmonella bacterial culture. The isolates were serotyped and evaluated as to antimicrobial susceptibility profile, extended-spectrum ?-lactamases (ESBL) production, and presence of virulence markers (invA, iroB, and spvC). RESULTS: A total of 5/3.2% children were infected by S. enterica, 3/60% by S. enterica Typhimurium, 1/20% by S. enterica Enteritidis and 1/20% S. enterica subsp. enterica serotype 8.20:z4,z23:-. Fecal leucocytes were detected in two out of five fecal specimens positive for S. enterica. Isolates from three children were resistant to nalidixic acid, nalidixic acid + chloramphenicol, and nalidixic acid + chloramphenicol + ampicillin. ESBL production was not detected. All samples presented invA and iroB genes. spvC marker was observed in isolates from two children infected by S. Typhimurium and S. Enteritidis. Conclusion: The results demonstrate that S. enterica infection is uncommon among children from our region. Furthermore, they indicate the need for periodic monitoring of bacterial antimicrobial susceptibility profile in order to establish suitable antimicrobial therapy when required.

Mireille Ângela Bernardes, Sousa; Edilberto Nogueira, Mendes; Francisco José, Penna; Luciano Amedée, Péret-Filho; Paula Prazeres, Magalhães.

106

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

Directory of Open Access Journals (Sweden)

Full Text Available Introduction: Acute infectious diarrhea is still regarded as a public health problem associated with a wide range of etiologic agents, from which Salmonella enterica is particularly worth mentioning inasmuch as it is a major cause of inflammatory diarrhea in both developed and developing countries. Objective: To assess the distribution of S. enterica among children with acute diarrhea in Belo Horizonte and to characterize bacterium isolates. Material and methods: The study group comprised a total of 157 children from low socioeconomic background. Stool samples were collected for leukocyte analysis and Salmonella bacterial culture. The isolates were serotyped and evaluated as to antimicrobial susceptibility profile, extended-spectrum ?-lactamases (ESBL production, and presence of virulence markers (invA, iroB, and spvC. RESULTS: A total of 5/3.2% children were infected by S. enterica, 3/60% by S. enterica Typhimurium, 1/20% by S. enterica Enteritidis and 1/20% S. enterica subsp. enterica serotype 8.20:z4,z23:-. Fecal leucocytes were detected in two out of five fecal specimens positive for S. enterica. Isolates from three children were resistant to nalidixic acid, nalidixic acid + chloramphenicol, and nalidixic acid + chloramphenicol + ampicillin. ESBL production was not detected. All samples presented invA and iroB genes. spvC marker was observed in isolates from two children infected by S. Typhimurium and S. Enteritidis. Conclusion: The results demonstrate that S. enterica infection is uncommon among children from our region. Furthermore, they indicate the need for periodic monitoring of bacterial antimicrobial susceptibility profile in order to establish suitable antimicrobial therapy when required.INTRODUÇÃO: A diarreia infecciosa aguda é considerada um problema de saúde pública associado a uma ampla gama de agentes etiológicos, entre os quais destaca-se Salmonella enterica, causa importante de diarreia inflamatória em países desenvolvidos e em desenvolvimento. OBJETIVOS: Avaliar a distribuição de S. enterica em crianças com diarreia aguda em Belo Horizonte e caracterizar as amostras isoladas. MATERIAL E MÉTODO: O grupo de estudo consistiu de 157 crianças de nível socioeconômico baixo. Espécimes fecais foram empregados para pesquisa de leucóc itos e cultivo de Salmonella. As amostras isoladas foram sorotipadas e submetidas à avaliação do perfil de suscetibilidade a antimicrobianos, da produção de betalactamases de amplo espectro (ESBL e da presença de marcadores de virulência (invA, iroB e spvC. RESULTADOS: Cinco/3,2% crianças apresentaram-se infectadas por S. enterica; três/60%, por S. enterica Typhimurium; uma/20%, por S. enterica Enteritidis; e uma/20%, por S. enterica subsp. enterica sorotipo 8,20:z4,z23:-. Leucócitos fecais foram detectados em dois dos cinco espécimes positivos para S. enterica. As amostras isoladas de três crianças apresentaram resistência a ácido nalidíxico, ácido nalidíxico + cloranfenicol e ácido nalidíxico + cloranfenicol + ampicilina. Nenhuma amostra produziu ESBL. Todas as amostras albergavam os genes invA e iroB. O marcador spvC foi observado em amostras isoladas de duas crianças infectadas por S. Typhimurium e S. Enteritidis. CONCLUSÃO: Os resultados demonstram que diarreia associada a S. enterica é raramente observada entre crianças da nossa região e indicam a necessidade de avaliação periódica do perfil de suscetibilidade a antimicrobianos da bactéria para orientar o estabelecimento de antibioticoterapia, quando indicada.

Mireille Ângela Bernardes Sousa

2013-02-01

107

Produção e purificação de anticorpos policlonais para Salmonella Enteritidis (Enterobacteriaceae Production and purification of polyclonal antibodies for Salmonella Enteritidis (Enterobacteriaceae  

Directory of Open Access Journals (Sweden)

Full Text Available O objetivo deste trabalho foi produzir e purificar anticorpos policlonais específicos para Salmonella Enteritidis (Enterobacteriaceae. O anti-soro foi produzido em coelhos, empregando-se flagelina purificada. O título e a especificidade foram determinados através do ensaio imunoenzimático - ELISA e a purificação por cromatografia de afinidade com sepharose Proteína A. As suspensões bacterianas foram cultivadas em cinco diferentes meios de cultura (infusão de cérebro coração - BHI, caldo tripticase soja, caldo lactosado, caldo nutriente - CN e água peptonada. Observou-se que dependendo do meio o título do anti-soro pode variar e os melhores resultados foram obtidos com BHI e CN. O anti-soro foi específico para Salmonella Enteritidis, apresentando porcentagens de reações cruzadas com Salmonella Typhimurium, Salmonella Infantis e Salmonella Newport de 16,0, 11,9 e 6,4%, respectivamente. Menores porcentagens foram obtidas com outras enterobactérias testadas. Esses resultados indicam a possibilidade da utilização desses anticorpos na padronização de ensaios imunológicos para a detecção de Salmonella EnteritidisThe purpose of this study was to produce and to purify specific polyclonal antibodies for Salmonella Enteritidis (Enterobacteriaceae. The anti-serum was raised in rabbits using purified flagelin. Anti-serum titer and specificity were determined by an immunoassay - ELISA and its purification was performed by sepharose protein A affinity chromatography. The bacteria suspensions were cultivated in five different media (brain heart infusion - BHI, tripticase soy broth, nutrient broth - NB, peptone water. Results have showed that anti-serum titers varied depending on which media type was used and BHI and NB media yielded the most significant results. The anti-serum produced was specific for Salmonella Enteritidis. Its cross-reactivity with Salmonella Thyphimurium, Salmonella Infantis and Salmonella Newport were 16.0, 11.9 and 6.4% respectively and lower percentages than those were observed in other Enterobacteriaceae species tested. Results indicate that these polyclonal antibodies may be used in standardization of immunological assays for Salmonella Enteritidis detection

Mario Augusto Ono

2002-04-01

108

Salmonella.org  

Science.gov (United States)

From the University of Illinois, Professor Stanley Maloy and Assistant Professor Rob Edwards' Web site Salmonella.org is dedicated to the study of the Salmonella bacteria genome. The site offers news and information on the bacteria's various strains, including everything from tips on preventing the infection to links to genomic sequencing data. Any Salmonella researcher or enthusiast will find this uncluttered and straightforward compilation useful.

Edwards, Rob.; Maloy, Stanley R.

2002-01-01

109

Envelope instability in DNA adenine methylase mutants of Salmonella enterica.  

Science.gov (United States)

Mutants of Salmonella enterica serovar Typhimurium lacking DNA adenine (Dam) methylase show reduced secretion of invasion effectors encoded in the Salmonella-pathogenicity island 1 (SPI-1). Concomitant with this alteration, a high number and quantity of extracellular proteins are detected in cultures of Dam(-) mutants. This study shows by subcellular fractionation analysis that the presence of numerous extracellular proteins in cultures of Dam(-) mutants is linked to an exacerbated release of membrane particulate material. The membrane 'leaky' phenotype and the impaired functionality of type III secretion systems were, however, unrelated since exacerbated release of proteins to the medium was evident in Dam(-) strains carrying mutations in either SPI-1 (invA, invJ) or flagellar (flhD) genes. This result supports the view that Dam methylation controls a plethora of cellular processes. Electron microscopy analysis demonstrated that the accumulation of membrane particulate material occurs preferentially as vesicles in stationary cultures of Dam(-) strains. In addition, a reduction in the relative amount of peptidoglycan-associated lipoprotein (PAL), TolB, OmpA and murein lipoprotein (Lpp) bound to peptidoglycan was observed in actively growing Dam(-) mutants. The existence of an envelope defect was further confirmed by the increased sensitivity to deoxycholate exhibited by Dam(-) mutants, mostly during exponential growth. Unexpectedly, lack of Dam methylation neither increased envelope instability nor impaired the association of PAL-Tol-Lpp proteins to the peptidoglycan in Escherichia coli. Accordingly, E. coli Dam(-) mutants did not show sensitivity to deoxycholate. Altogether, these results indicate that, besides its role in modulating the secretion of effectors by the SPI-1-encoded type III apparatus, Dam methylation controls cell envelope integrity in S. enterica. PMID:11932461

Pucciarelli, M Graciela; Prieto, Ana I; Casadesús, Josep; García-del Portillo, Francisco

2002-04-01

110

A comparative study on the pathogenesis of egg contamination by different serotypes of Salmonella.  

Science.gov (United States)

Salmonella enterica serotype Enteritidis is the predominant serotype associated with egg-borne salmonellosis in humans. Apparently this serotype possesses particular characteristics that increase its chance to contaminate eggs. To identify these characteristics, two Salmonella serotype Enteritidis strains as well as one strain of each of the serotypes Salmonella Typhimurium, Salmonella Heidelberg, Salmonella Virchow and Salmonella Hadar strain were used to examine different aspects related to egg contamination. After an intravenous infection of laying hens, it was observed that the ability of the serotype Enteritidis strains to colonize the reproductive organs was significantly higher compared with the Salmonella Heidelberg, Salmonella Virchow and Salmonella Hadar strains but not with the Salmonella Typhimurium strain. Inoculating low numbers of the different Salmonella serotypes in egg albumen at 42 degrees C demonstrated that the growth of the strains belonging to the Salmonella serotypes Virchow and Hadar was seriously repressed. The other serotypes, however, survived in albumen for 24 h. Furthermore, using two different specifically designed egg infection models, it was shown that all strains used in this study were able to penetrate into and multiply inside the yolk at 25 degrees C. These findings indicate that the ability to grow in eggs post lay is not specific for the serotype Enteritidis. In conclusion, comparing strains belonging to different Salmonella serotypes has revealed that most probably a preferential colonization of the reproductive organs and an enhanced survival at 42 degrees C allows the serotype Enteritidis to contaminate eggs. PMID:18622856

Gantois, I; Eeckhaut, V; Pasmans, F; Haesebrouck, F; Ducatelle, R; Van Immerseel, F

2008-08-01

111

The Salmonella enterica subspecies I specific centisome 7 genomic island encodes novel protein families present in bacteria living in close contact with eukaryotic cells  

DEFF Research Database (Denmark)

We have determined the genetic structure of the Salmonella enterica centisome 7 genomic island (SCI) located at the aspV loci in S. enterica subspecies I strains. The 47-kb long genomic island encodes 37 putative proteins, including the previously described saf fimbrial operon and the sinR transcriptional regulator. Other open reading frames (designated sci A to Z) in the island encode putative proteins with homologies to virulence-associated proteins in a number of Gram-negative bacteria such as Pseudomonas aeruginosa, Yersinia pestis and enterohemorrhagic Escherichia coli, bacteria that have the ability to interact with and manipulate eukaryotic cells. The Sci proteins have putative cytoplasmic, periplasmic and outer membrane localizations pointing to a role in extracellular processes such as secretion or organelle biosynthesis. The genes encoding Sci-like proteins are clustered in all sequenced bacterial genomes available in the databases and a core set can be defined by the presence of genes encoding proteins with similarity to the SciB, C, G, H, I, O proteins. The SCI genomic island DNA sequences are restricted to Salmonella strains belonging to S. enterica subspecies I and deletion of the entire island affects the ability of the organisms to enter eukaryotic cells.

Folkesson, Anders; Löfdahl, Sven

2002-01-01

112

SYBR®Green qPCR Salmonella detection system allowing discrimination at the genus, species and subspecies levels.  

Science.gov (United States)

In this work, a three-level Salmonella detection system based on a combination of seven SYBR®Green qPCR was developed. This detection system discriminates Salmonella at the genus, species and subspecies levels using a single 96-well plate. The SYBR®Green qPCR assays target the invA, rpoD, iroB and safC genes, as well as the STM0296 locus, putatively coding for a cytoplasmic protein. This study includes the design of primer pairs, in silico and in situ selectivity, sensitivity, repeatability and reproducibility evaluations of the seven SYBR®Green qPCR assays. Each detection level displayed a selectivity of 100 %. This combinatory SYBR®Green qPCR system was also compared with three commercially available Salmonella qPCR detection kits. This comparison highlighted the importance of using a multi-gene detection system to be able to detect every target strain, even those with deletion or mutation of important genes. PMID:24113820

Barbau-Piednoir, Elodie; Bertrand, Sophie; Mahillon, Jacques; Roosens, Nancy H; Botteldoorn, Nadine

2013-11-01

113

MÉTODOS DE EXTRAÇÃO DE DNA PARA A DETECÇÃO DE Salmonella EM OVOS DE GALINHAS, COM E SEM CASCA, ATRAVÉS DA REAÇÃO EM CADEIA PELA POLIMERASE DNA EXTRACTION METHODS FOR Salmonella DETECTION IN CHICKEN EGGS, INSHELL AND OUTSHELL, BY POLIMERASE CHAIN REACTION  

Directory of Open Access Journals (Sweden)

Full Text Available O diagnóstico microbiológico de Salmonella sp em amostras de alimentos é demorado, com cinco diferentes etapas, levando cerca de 120 horas até o resultado final. A utilização da técnica da Reação em Cadeia pela Polimerase (PCR pode diminuir esse período, porém sofre influência de substâncias presentes na amostra que afetam a reação. O objetivo deste trabalho foi comparar dois métodos de extração de DNA, a extração por tratamento térmico e a pelo fenol-clorofórmio, em amostras de 100 ovos de galinhas domésticas artificialmente contaminados com uma cepa de Salmonella enterica sorovar typhimurium em fase estacionária. O material obtido com as extrações foi submetido à PCR, utilizando-se um par de iniciadores que amplificam um fragmento de 284pb do gene InvA de Salmonella sp. Comparando os métodos de extração, observou-se uma diferença na capacidade de detecção de 12% a favor do método do fenol-clorofórmio, quando a extração foi realizada a partir do ovo com casca. No momento em que a mesma metodologia foi usada apenas com a parte interna dos ovos, essa diferença subiu para 26% o que foi significativo (PThe Salmonella sp detection in feed samples is time consuming, it has five stages and requires 120 hours for final results. The use of polimerase chain reaction technique can reduce this time considerably, however it can be affected by substances from the sample. This study had the objective of comparing two methods of DNA extraction, by heating process and by phenol-chloroform in samples of 100 chicken eggs experimentally infected with a sample of Salmonella enterica sorovar typhimurium in stationary phase. After the two extraction methods a PCR was done using a pair of oligonucleotides that amplifies a fragment of 284pb in the InvA gene de Salmonella sp. Comparing the extraction methods it was noted a difference of 12% favorably to the phenol-chloroform method when the extraction was done from eggs with shell. The same method using just the egg internal part resulted in a difference of 26% with high significance (P<0.003, showing that the method used was determinant to improve the technique efficiency. However, comparing the positive percentage independent of the DNA extraction method a significant difference (P<0.047 was noted for outshell eggs. This fact suggests that for Salmonella egg analysis, only the egg internal part should be used because the shell can determine interference in technique.

Maristela Lovato Flôres

2001-04-01

114

Salmonella enterica serotype 4,5,12:i:-, an emerging Salmonella serotype that represents multiple distinct clones.  

Science.gov (United States)

The prevalence, among human clinical cases, of Salmonella enterica serotype 4,5,12:i:-, a serotype antigenically similar to Salmonella enterica serotype Typhimurium but lacking second-phase flagellar antigens, has increased considerably over the last 10 years. To probe the evolution and ecology of this emerging serotype, we characterized 190 Salmonella isolates initially classified as Salmonella serotypes 4,5,12:i:- (n = 90) and Typhimurium (n = 100) and obtained from various sources in the United States and Spain. These isolates were characterized into six sequence types (determined by multilocus sequence typing [MLST]) and 79 pulsed-field gel electrophoresis types. The majority of Salmonella serotype 4,5,12:i:- and Typhimurium isolates (85 and 84 isolates, respectively) represented a single MLST type. Existing genome information revealed different genome deletions (which included genes responsible for phase 2 flagellum expression) in four Spanish Salmonella serotype 4,5,12:i:- isolates and one U.S. Salmonella serotype 4,5,12:i:- isolate. Fifty-nine isolates of both serotypes, representing different sources and geographical locations as well as different molecular subtypes, were thus screened for the presence of six genes and one specific region, all of which were previously found to show variable presence among Salmonella serotype 4,5,12:i:- and Typhimurium strains. All Salmonella serotype 4,5,12:i:- isolates lacked the phase 2 flagella genes fljA and fljB, which were present in all Salmonella serotype Typhimurium isolates. While all Spanish Salmonella serotype 4,5,12:i:- isolates carried the same deletion surrounding fljAB, all but two U.S. isolates showed a different genomic deletion; the two atypical U.S. isolates represented the "Spanish" deletion genotype and a unique deletion genotype. Salmonella serotype 4,5,12:i:- thus appears to represent at least two common clones, which cannot easily be differentiated with standard diagnostic procedures. PMID:19741087

Soyer, Y; Moreno Switt, A; Davis, M A; Maurer, J; McDonough, P L; Schoonmaker-Bopp, D J; Dumas, N B; Root, T; Warnick, L D; Gröhn, Y T; Wiedmann, M

2009-11-01

115

Identification of Salmonella with the O-1 bacteriophage.  

Science.gov (United States)

The O-1 bacteriophage test of Cherry et al. (1954) for the presumptive identification of salmonellae in the diagnostic laboratory was investigated. A phage lysate with a titer of 10(12) plaque-forming units per ml was found to be optimal. This preparation lysed 98.2% of Salmonella strains tested, while maintaining its high specificity for salmonellae. Gram-negative organisms other than salmonellae were resistant to the O-1 phage; however, 5.9% of Escherichia coli strains tested were susceptible. The O-1 phage test is a simple, rapid, inexpensive, sensitive, and specific procedure for the identification of salmonellae in the diagnostic laboratory. A presumptive identification is obtained 1 day earlier than with conventional biochemical tests. PMID:4609209

Welkos, S; Schreiber, M; Baer, H

1974-10-01

116

Detection of Salmonella spp, Salmonella Enteritidis and Typhimurium in naturally infected broiler chickens by a multiplex PCR-based assay  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english The presence of Salmonella in the intestinal tract, on the chickens skin and among their feathers, may cause carcasses contamination during slaughtering and processing and possibly it is responsible by the introduction of this microorganism in the slaughterhouses. A rapid method to identify and moni [...] tor Salmonella and their sorovars in farm is becoming necessary. A pre-enriched multiplex polymerase chain reaction (m-PCR) assay employing specific primers was developed and used to detect Salmonella at the genus level and to identify the Salmonella enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterica serovar Typhimurium (S. Typhimurium) in broiler chicken swab samples. The method was validated by testing DNA extract from 90 fresh culture cloacal swab samples from poultry chicken cultured in phosphate buffer peptone water at 37 ºC for 18 h. The final results showed the presence of Salmonella spp. in 25% of samples, S. Enteritidis was present in 12% of the Salmonella-positive samples and S. Typhimurium in 3% of the samples. The m-PCR assay developed in this study is a specific and rapid alternative method for the identification of Salmonella spp. and allowed the observation of specific serovar contamination in the field conditions within the locations where these chickens are typically raised.

F.G., Paião; L.G.A., Arisitides; L.S., Murate; G.T., Vilas-Bôas; L.A., Vilas-Boas; M., Shimokomaki.

117

Bacteriophage therapy to reduce salmonella colonization of broiler chickens.  

Science.gov (United States)

Acute enteric infections caused by salmonellas remain a major public health burden worldwide. Poultry, particularly chickens, are known to be the main reservoir for this zoonotic pathogen. Although some progress has been made in reducing Salmonella colonization of broiler chickens by using biosecurity and antimicrobials, it still remains a considerable problem. The use of host-specific bacteriophages as a biocontrol is one possible intervention by which Salmonella colonization could be reduced. A total of 232 Salmonella bacteriophages were isolated from poultry farms, abattoirs, and wastewater in 2004 and 2005. Three phages exhibiting the broadest host ranges against Salmonella enterica serotypes Enteritidis, Hadar, and Typhimurium were characterized further by determining their morphology and lytic activity in vitro. These phages were then administered in antacid suspension to birds experimentally colonized with specific Salmonella host strains. The first phage reduced S. enterica serotype Enteritidis cecal colonization by > or = 4.2 log10 CFU within 24 h compared with controls. Administration of the second phage reduced S. enterica serotype Typhimurium by > or = 2.19 log10 CFU within 24 h. The third bacteriophage was ineffective at reducing S. enterica serotype Hadar colonization. Bacteriophage resistance occurred at a frequency commensurate with the titer of phage being administered, with larger phage titers resulting in a greater proportion of resistant salmonellas. The selection of appropriate bacteriophages and optimization of both the timing and method of phage delivery are key factors in the successful phage-mediated control of salmonellas in broiler chickens. PMID:17526794

Atterbury, R J; Van Bergen, M A P; Ortiz, F; Lovell, M A; Harris, J A; De Boer, A; Wagenaar, J A; Allen, V M; Barrow, P A

2007-07-01

118

Real-time reverse-transcriptase--polymerase chain reaction for Salmonella enterica detection from jalapeño and serrano peppers.  

Science.gov (United States)

Outbreaks of Salmonella linked to fresh produce emphasize the need for rapid detection methods to curb the spread of foodborne pathogens. Reverse-transcriptase-polymerase chain reaction (RT-PCR) detects the presence of mRNA (shorter half-life than DNA), with greater potential of detecting viable pathogens. Real-time RT-PCR eliminates the need for gel electrophoresis and significantly enhances the speed of detection (5 days). The objectives of this research were to apply real-time SYBR Green I-based RT-PCR to detect Salmonella from jalapeño and serrano peppers spiked with low and high inocula of Salmonella. Inoculated and uninoculated peppers were rinsed with water and dried under ultraviolet light for 10 min. Approximately 25 g peppers was inoculated with 10(8) to 10(1) colony forming units (CFU) of Salmonella enterica serovar Typhimurium in a stomacher bag and hand massaged in sterile 0.05 M glycine-0.14 M saline buffer (0.05% Tween, 3% beef extract) for optimal recovery of bacteria. A short preenrichment step of 6 h in buffered peptone water was needed for the detection of low inocula (10(4) CFU/25 g). One-milliliter portions of the extracts were serially diluted, plated on XLT4 agar, and used for RNA extraction with the Qiagen RNeasy Mini Kit. RT-PCR was carried out using SYBR Green I one-step RT-PCR with previously described invA gene primers and an internal amplification control. Detection limits were 10(4) CFU/25 g (approximately 10(2) CFU/g) and 10(7) CFU/25 g (approximately 10(5) CFU/g) Salmonella from enriched and unenriched inoculated peppers, respectively. Even though this method included a 6-h incubation period, the results were still obtainable in 1 day. This method shows promise for applications in routine surveillance and during outbreaks. PMID:19911882

Miller, Nathan D; Draughon, Frances Ann; D'Souza, Doris H

2010-04-01

119

Evaluation of an indirect ELISA for the detection of Salmonella in chicken meat Avaliação de um ELISA indireto para detecção de Salmonella em carne de frango  

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Full Text Available In this work, an indirect ELISA based on a monoclonal antibody (MAb specific for an outer membrane protein of Salmonellaenterica serovar Enteritidis was used for detection of Salmonella in 154 samples of chicken meat. Its efficiency was determined through comparison with the results obtained from the conventional method. The prevalence of samples contaminated with Salmonella was 23% with the conventional culture method, and 26% with the ELISA. From thirty-five samples positive for Salmonella by the conventional method, 33 were also positive by ELISA. Seven other samples were only positive in the ELISA. Comparison of the results obtained in the two methods showed an ELISA sensitivity and specificity of 94%, and positive and negative predictive values of 82% and 98% respectively. The serotyping of the isolates revealed 31 Salmonella enterica serovar Enteritidis, 2 Salmonella enterica serovar Heidelberg, 1 Salmonella enterica serovar Choleraesuis and 1 Salmonella enterica sorovar 6,7:-:-.Neste trabalho, um ELISA indireto baseado em um anticorpo monoclonal (MAb especifico para proteína de membrane externa de Salmonellaenterica serovar Enteritidis foi usado para detecção de Salmonella em 154 amostras de carne de frango. Sua eficiência foi determinada através de comparação com os resultados obtidos pela metodologia convencional. A prevalência de amostras contaminadas com Salmonella foi de 23% pelo método de cultivo convencional, e 26% pelo ELISA. De 35 amostras positivas para Salmonella pela metodologia convencional, 32 também foram positivas no ELISA. Outras sete amostras foram positivas somente no ELISA. Comparando os resultados obtidos nos dois métodos, o ELISA demonstrou sensibilidade e especificidade de 94%, e valor preditivo positivo e negativo de 82% e 98% respectivamente. A sorotipagem dos isolados revelou 31 Salmonella enterica serovar Enteritidis, 2 Salmonella enterica serovar Heidelberg, 1 Salmonella enterica serovar Choleraesuis e 1 Salmonella enterica sorovar 6,7:-:-.

Andréa dos Santos Schneid

2006-09-01

120

Evaluation of an indirect ELISA for the detection of Salmonella in chicken meat / Avaliação de um ELISA indireto para detecção de Salmonella em carne de frango  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese Neste trabalho, um ELISA indireto baseado em um anticorpo monoclonal (MAb) especifico para proteína de membrane externa de Salmonellaenterica serovar Enteritidis foi usado para detecção de Salmonella em 154 amostras de carne de frango. Sua eficiência foi determinada através de comparação com os resu [...] ltados obtidos pela metodologia convencional. A prevalência de amostras contaminadas com Salmonella foi de 23% pelo método de cultivo convencional, e 26% pelo ELISA. De 35 amostras positivas para Salmonella pela metodologia convencional, 32 também foram positivas no ELISA. Outras sete amostras foram positivas somente no ELISA. Comparando os resultados obtidos nos dois métodos, o ELISA demonstrou sensibilidade e especificidade de 94%, e valor preditivo positivo e negativo de 82% e 98% respectivamente. A sorotipagem dos isolados revelou 31 Salmonella enterica serovar Enteritidis, 2 Salmonella enterica serovar Heidelberg, 1 Salmonella enterica serovar Choleraesuis e 1 Salmonella enterica sorovar 6,7:-:-. Abstract in english In this work, an indirect ELISA based on a monoclonal antibody (MAb) specific for an outer membrane protein of Salmonellaenterica serovar Enteritidis was used for detection of Salmonella in 154 samples of chicken meat. Its efficiency was determined through comparison with the results obtained from t [...] he conventional method. The prevalence of samples contaminated with Salmonella was 23% with the conventional culture method, and 26% with the ELISA. From thirty-five samples positive for Salmonella by the conventional method, 33 were also positive by ELISA. Seven other samples were only positive in the ELISA. Comparison of the results obtained in the two methods showed an ELISA sensitivity and specificity of 94%, and positive and negative predictive values of 82% and 98% respectively. The serotyping of the isolates revealed 31 Salmonella enterica serovar Enteritidis, 2 Salmonella enterica serovar Heidelberg, 1 Salmonella enterica serovar Choleraesuis and 1 Salmonella enterica sorovar 6,7:-:-.

Andréa dos Santos, Schneid; Kelly Lameiro, Rodrigues; Davi, Chemello; Eduardo Cesar, Tondo; Marco Antônio Zacchia, Ayub; José Antonio Guimarães, Aleixo.

 
 
 
 
121

Salmonella Diagnosis and Treatment  

Science.gov (United States)

... Outbreaks Multidrug-Resistant Salmonella Heidelberg Infections Linked to Foster Farms Brand Chicken Recall & Advice to Consumers Case Count Maps Epi Curves Signs & Symptoms Key Resources Timeline of Events Montevideo ...

122

MÉTODOS DE EXTRAÇÃO DE DNA PARA A DETECÇÃO DE Salmonella EM OVOS DE GALINHAS, COM E SEM CASCA, ATRAVÉS DA REAÇÃO EM CADEIA PELA POLIMERASE / DNA EXTRACTION METHODS FOR Salmonella DETECTION IN CHICKEN EGGS, INSHELL AND OUTSHELL, BY POLIMERASE CHAIN REACTION  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese O diagnóstico microbiológico de Salmonella sp em amostras de alimentos é demorado, com cinco diferentes etapas, levando cerca de 120 horas até o resultado final. A utilização da técnica da Reação em Cadeia pela Polimerase (PCR) pode diminuir esse período, porém sofre influência de substâncias presen [...] tes na amostra que afetam a reação. O objetivo deste trabalho foi comparar dois métodos de extração de DNA, a extração por tratamento térmico e a pelo fenol-clorofórmio, em amostras de 100 ovos de galinhas domésticas artificialmente contaminados com uma cepa de Salmonella enterica sorovar typhimurium em fase estacionária. O material obtido com as extrações foi submetido à PCR, utilizando-se um par de iniciadores que amplificam um fragmento de 284pb do gene InvA de Salmonella sp. Comparando os métodos de extração, observou-se uma diferença na capacidade de detecção de 12% a favor do método do fenol-clorofórmio, quando a extração foi realizada a partir do ovo com casca. No momento em que a mesma metodologia foi usada apenas com a parte interna dos ovos, essa diferença subiu para 26% o que foi significativo (P Abstract in english The Salmonella sp detection in feed samples is time consuming, it has five stages and requires 120 hours for final results. The use of polimerase chain reaction technique can reduce this time considerably, however it can be affected by substances from the sample. This study had the objective of comp [...] aring two methods of DNA extraction, by heating process and by phenol-chloroform in samples of 100 chicken eggs experimentally infected with a sample of Salmonella enterica sorovar typhimurium in stationary phase. After the two extraction methods a PCR was done using a pair of oligonucleotides that amplifies a fragment of 284pb in the InvA gene de Salmonella sp. Comparing the extraction methods it was noted a difference of 12% favorably to the phenol-chloroform method when the extraction was done from eggs with shell. The same method using just the egg internal part resulted in a difference of 26% with high significance (P

Maristela Lovato, Flôres; Vladimir Pinheiro do, Nascimento; Ivonyr Irene Troglio Abdel, Kader; Luciana Ruschel dos, Santos; Alexandre Pontes, Pontes; Carlos Tadeu Pippi, Salle; Rui Fernando Felix, Lopes.

123

MÉTODOS DE EXTRAÇÃO DE DNA PARA A DETECÇÃO DE Salmonella EM OVOS DE GALINHAS, COM E SEM CASCA, ATRAVÉS DA REAÇÃO EM CADEIA PELA POLIMERASE / DNA EXTRACTION METHODS FOR Salmonella DETECTION IN CHICKEN EGGS, INSHELL AND OUTSHELL, BY POLIMERASE CHAIN REACTION  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese O diagnóstico microbiológico de Salmonella sp em amostras de alimentos é demorado, com cinco diferentes etapas, levando cerca de 120 horas até o resultado final. A utilização da técnica da Reação em Cadeia pela Polimerase (PCR) pode diminuir esse período, porém sofre influência de substâncias presen [...] tes na amostra que afetam a reação. O objetivo deste trabalho foi comparar dois métodos de extração de DNA, a extração por tratamento térmico e a pelo fenol-clorofórmio, em amostras de 100 ovos de galinhas domésticas artificialmente contaminados com uma cepa de Salmonella enterica sorovar typhimurium em fase estacionária. O material obtido com as extrações foi submetido à PCR, utilizando-se um par de iniciadores que amplificam um fragmento de 284pb do gene InvA de Salmonella sp. Comparando os métodos de extração, observou-se uma diferença na capacidade de detecção de 12% a favor do método do fenol-clorofórmio, quando a extração foi realizada a partir do ovo com casca. No momento em que a mesma metodologia foi usada apenas com a parte interna dos ovos, essa diferença subiu para 26% o que foi significativo (P Abstract in english The Salmonella sp detection in feed samples is time consuming, it has five stages and requires 120 hours for final results. The use of polimerase chain reaction technique can reduce this time considerably, however it can be affected by substances from the sample. This study had the objective of comp [...] aring two methods of DNA extraction, by heating process and by phenol-chloroform in samples of 100 chicken eggs experimentally infected with a sample of Salmonella enterica sorovar typhimurium in stationary phase. After the two extraction methods a PCR was done using a pair of oligonucleotides that amplifies a fragment of 284pb in the InvA gene de Salmonella sp. Comparing the extraction methods it was noted a difference of 12% favorably to the phenol-chloroform method when the extraction was done from eggs with shell. The same method using just the egg internal part resulted in a difference of 26% with high significance (P

Maristela Lovato, Flôres; Vladimir Pinheiro do, Nascimento; Ivonyr Irene Troglio Abdel, Kader; Luciana Ruschel dos, Santos; Alexandre Pontes, Pontes; Carlos Tadeu Pippi, Salle; Rui Fernando Felix, Lopes.

2001-04-01

124

Salmonella enterica: A surprisingly well adapted intracellular lifestyle  

Directory of Open Access Journals (Sweden)

Full Text Available The infectious intracellular lifestyle of Salmonella enterica relies on the adaptation to nutritional conditions within the Salmonella-containing vacuole (SCV in host cells. We summarize latest results on metabolic requirements for Salmonella during infection. This includes intracellular phenotypes of mutant strains based on metabolic modeling and experimental tests, isotopologue profiling using 13C-compounds in intracellular Salmonella, and complementation of metabolic defects for attenuated mutant strains towards a comprehensive understanding of the metabolic requirements of the intracellular lifestyle of Salmonella. Helpful for this are also genomic comparisons. We outline further recent studies and which analyses of intracellular phenotypes and improved metabolic simulations were done and comment on technical required steps as well as progress involved in the iterative refinement of metabolic flux models, analyses of mutant phenotypes and isotopologue analyses. Salmonella lifestyle is well adapted to the SCV and its specific metabolic requirements. Salmonella metabolism adapts rapidly to SCV conditions, the metabolic generalist Salmonella is quite successful in host infection.

ThomasDandekar

2012-05-01

125

Salmonella enterica: a surprisingly well-adapted intracellular lifestyle.  

Science.gov (United States)

The infectious intracellular lifestyle of Salmonella enterica relies on the adaptation to nutritional conditions within the Salmonella-containing vacuole (SCV) in host cells. We summarize latest results on metabolic requirements for Salmonella during infection. This includes intracellular phenotypes of mutant strains based on metabolic modeling and experimental tests, isotopolog profiling using (13)C-compounds in intracellular Salmonella, and complementation of metabolic defects for attenuated mutant strains towards a comprehensive understanding of the metabolic requirements of the intracellular lifestyle of Salmonella. Helpful for this are also genomic comparisons. We outline further recent studies and which analyses of intracellular phenotypes and improved metabolic simulations were done and comment on technical required steps as well as progress involved in the iterative refinement of metabolic flux models, analyses of mutant phenotypes, and isotopolog analyses. Salmonella lifestyle is well-adapted to the SCV and its specific metabolic requirements. Salmonella metabolism adapts rapidly to SCV conditions, the metabolic generalist Salmonella is quite successful in host infection. PMID:22563326

Dandekar, Thomas; Astrid, Fieselmann; Jasmin, Popp; Hensel, Michael

2012-01-01

126

Meta-analysis of Chicken – Salmonella infection experiments  

Directory of Open Access Journals (Sweden)

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.

te Pas Marinus FW

2012-04-01

127

Multiplex T-RFLP allows for increased target number and specificity: detection of Salmonella enterica and six species of Listeria in a single test.  

Science.gov (United States)

A multiplex T-RFLP test was developed to detect and identify Salmonella enterica and all six species of Listeria inoculated into milk at minimal levels. Extensive in silico analysis was used to design a fifteen-primer, six-amplimer methodology and in vitro application showed target organism DNA, when amplified individually, yielded the predicted terminal restriction fragments (TRFs) following digestion. Non-target organisms were either not-amplified or yielded TRFs which did not interfere with target identification. Multiple target DNA analysis gave over 86% detection of total TRFs predicted, and this was improved to over 90% detection of total TRFs predicted when only two target DNA extracts were combined analysed. Co-inoculation of milk with five strains each of the target species of S. enterica and L. monocytogenes, along with five strains of the non-target species E. coli was followed by enrichment in SEL medium for M-TRFLP analysis. This allowed for detection of both target species in all samples, with detection of one S. enterica and two Listeria TRFs in all cases, and detection of a second S. enterica TRF in 91% of cases. This was from an initial inoculum of <5 cfu per 25 ml milk with a background of competing E. coli present, and gave a result from sampling of under 20 hours. The ability to increase target species number without loss of sensitivity means that extensive screening can be performed at reduced cost due to a reduction in the number of tests required. PMID:22937073

Elliott, Geoffrey N; Thomas, Nadine; Macrae, Marion; Campbell, Colin D; Ogden, Iain D; Singh, Brajesh K

2012-01-01

128

Biofilms promote survival and virulence of Salmonella enterica sv. Tennessee during prolonged dry storage and after passage through an in vitro digestion system.  

Science.gov (United States)

Salmonella enterica serotypes have been linked to outbreaks associated with low water activity foods. While the biofilm-forming abilities of Salmonella improve its survival during thermal processing and sanitation it is unclear whether biofilms enhance survival to desiccation and gastric stresses. The purpose of this study was to quantify the effect of physiological state (planktonic versus biofilm) and prior exposure to desiccation and storage in dry milk powder on Salmonella survival and gene expression after passage through an in vitro digestion model. Planktonic cells of Salmonella enterica serotype Tennessee were deposited onto membranes while biofilms were formed on glass beads. The cells were subsequently dried at room temperature and stored in dried milk powder (a(w)=0.3) for up to 30 days. Salmonella survival was quantified by serial dilution onto Brilliant Green Agar before desiccation, after desiccation, after 1-day storage and after 30-day storage. At each sampling period both physiological states were tested for survival through a simulated gastrointestinal system. RNA was extracted at the identical time points and Quantitative Real-Time PCR was used to determine relative expression for genes associated with stress response (rpoS, otsB), virulence (hilA, invA, sipC) and a housekeeping gene 16S rRNA. The physiological state and length of storage affected the survival and gene expression of Salmonella within the desiccated milk powder environment and after passage through an in vitro digestion system (pviable but non-culturable state. The increased expression of stress response genes rpoS and otsB correlated with survival, indicating cross-protection to low water activity and acid stress. Increased expression of virulence-associated genes was seen in cells exposed to dry storage for short periods, however the largest amount of expression occurred in biofilm cells stored for 30 days at aw 0.3, suggesting increased virulence potential. PMID:23454816

Aviles, Bryan; Klotz, Courtney; Eifert, Joseph; Williams, Robert; Ponder, Monica

2013-04-01

129

Assessing risk profiles for Salmonella serotypes in breeding pig operations in Portugal using a Bayesian hierarchical model  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Abstract Background The EU Regulation No 2160/2003 imposes a reduction in the prevalence of Salmonella in pigs. The efficiency of control programmes for Salmonella in pigs, reported among the EU Member States, varies and definitive eradication seems very difficult. Control measures currently recommended for Salmonella are not serotype-specific. Is it possible that the risk factors for different Salmonella serotypes are different? The aim of...

Correia-Gomes Carla; Economou Theodoros; Mendonça Denisa; Vieira-Pinto Madalena; Niza-Ribeiro João

2012-01-01

130

A critical review of Salmonella Typhimurium infection in laying hens.  

Science.gov (United States)

Salmonella Typhimurium has been reported to contaminate egg production across the world, but where Salmonella Enteritidis is endemic it is this latter serovar that dominates egg-borne salmonellosis. However, Salmonella Typhimurium is a major food-borne pathogen so it is important to understand how it can impact the microbiological safety of eggs and what serovar-specific control strategies may be appropriate in the future as control over Salmonella Enteritidis continues to improve. To that end, the present review examines the published literature on Salmonella Typhimurium in laying hens and eggs, with particular reference to comparative studies examining different serovars. Experimentally Salmonella Enteritidis is more often isolated from egg contents and seems to adhere better to reproductive tract mucosa, whilst Salmonella Typhimurium appears to provoke a more intense tissue pathology and immune response, and flock infections are more transient. However, it is observed in many cases that the present body of evidence does not identify clear differences between specific behaviours of the serovars Typhimurium and Enteritidis, whether in laying hens, in their eggs, or in the laying environment. It is concluded that further long-term experimental and natural infection studies are needed in order to generate a clearer picture. PMID:21879803

Wales, A D; Davies, R H

2011-10-01

131

Excreción fecal de Salmonella Albany, su aislamiento en la ración alimenticia y repercusión en el estado de salud de un ocelote (Leopardus pardalis) en cautiverio / Fecal excretion of Salmonella Albany, its isolation in the diet and health repercussion on an ocelot (Leopardus pardalis) in captivity  

Scientific Electronic Library Online (English)

Full Text Available SciELO Mexico | Language: Spanish Abstract in spanish Los serotipos de Salmonella especie enterica son los responsables del 99% de las salmonelosis en humanos y animales, en particular, Salmonella enterica serovariedad Albany se ha identificado en canales de pollo, por lo que representa un riesgo para la salud humana y animal. Se aisló Salmonella enter [...] ica serovariedad Albany a partir de heces de un ocelote macho (Leopardus pardalis), cautivo en el zoológico de Culiacán, Sinaloa, México, y de pollo crudo (alimento del felino). El patrón por electroforesis en campo pulsado (PFGE) con la enzima Xba I fue idéntico en ambos aislados, lo que indica que la fuente de infección fue el pollo crudo. Cinco meses después de haber aislado las bacterias de las heces, se realizó estudio post mortem del felino anteriormente mencionado, y se observó macroscópicamente: enterocolitis hemorrágica severa y fibrosis renal; y microscópicamente: necrosis de vellosidades y de criptas e infiltrado mononuclear linfocitario severo en íleon, además nefritis intersticial severa multifocal y fibrosis en riñón. A partir de muestras intestinales se amplificó el gen invA que confirma la infección por Salmonella. Los diagnósticos microbiológico, molecular e histopatológico sugieren que la muerte del felino se debió a la infección causada por la ingesta de pollo crudo contaminado con Salmonella enterica serovariedad Albany. Este caso clínico confirma la importancia que tienen los animales que excretan Salmonella vía fecal y describe la relación epidemiológica-molecular de los aislamientos obtenidos de heces y alimento, lo que permitió esclarecer la fuente primaria de infección. Abstract in english Salmonella enterica serotypes are 99% responsible for salmonellosis in human and animals, especially Salmonella enterica serovar Albany that has been identified in chicken carcass representing a risk for human and animal health. Salmonella enterica serovar Albany was isolated from the feces of a mal [...] e ocelot (Leopardus pardalis), at the zoo in Culiacan, Sinaloa, Mexico, and from raw chicken (feline's diet). The pulsed-field gel electrophoresis pattern (PFGE) generated by Xba I enzyme was identical in both isolates, indicating that the source of infection was the raw chicken. Five months after having isolated the bacteria from the feces, a post mortem study was carried out on the feline. Macroscopically, severe hemorrhagic enterocolitis and renal fibrosis was observed and microscopically, there was evidence of severe mononuclear lymphocytic infiltration in the ileum, as well as necrosis of intestinal villi and crypts, besides severe multifocal interstitial nephritis and fibrosis in both kidneys. The invA gene was amplified from intestinal samples confirming an infection by Salmonella. The microbiologic, molecular and histopathology diagnoses suggest that death of the feline was caused by ingestion of raw chicken contaminated with Salmonella enterica serovar Albany. This clinical case highlights the importance of persistent fecal Salmonella shedding animals and describes the molecular epidemiological relationships of isolates from feces and food, which allowed to find the primary source of infection.

Gabriela, Silva-Hidalgo; Héctor Samuel, López-Moreno; Vianney F., Ortiz-Navarrete; Felipe, Juárez-Barranco; Martín, López-Valenzuela.

2012-03-01

132

Excreción fecal de Salmonella Albany, su aislamiento en la ración alimenticia y repercusión en el estado de salud de un ocelote (Leopardus pardalis) en cautiverio / Fecal excretion of Salmonella Albany, its isolation in the diet and health repercussion on an ocelot (Leopardus pardalis) in captivity  

Scientific Electronic Library Online (English)

Full Text Available SciELO Mexico | Language: Spanish Abstract in spanish Los serotipos de Salmonella especie enterica son los responsables del 99% de las salmonelosis en humanos y animales, en particular, Salmonella enterica serovariedad Albany se ha identificado en canales de pollo, por lo que representa un riesgo para la salud humana y animal. Se aisló Salmonella enter [...] ica serovariedad Albany a partir de heces de un ocelote macho (Leopardus pardalis), cautivo en el zoológico de Culiacán, Sinaloa, México, y de pollo crudo (alimento del felino). El patrón por electroforesis en campo pulsado (PFGE) con la enzima Xba I fue idéntico en ambos aislados, lo que indica que la fuente de infección fue el pollo crudo. Cinco meses después de haber aislado las bacterias de las heces, se realizó estudio post mortem del felino anteriormente mencionado, y se observó macroscópicamente: enterocolitis hemorrágica severa y fibrosis renal; y microscópicamente: necrosis de vellosidades y de criptas e infiltrado mononuclear linfocitario severo en íleon, además nefritis intersticial severa multifocal y fibrosis en riñón. A partir de muestras intestinales se amplificó el gen invA que confirma la infección por Salmonella. Los diagnósticos microbiológico, molecular e histopatológico sugieren que la muerte del felino se debió a la infección causada por la ingesta de pollo crudo contaminado con Salmonella enterica serovariedad Albany. Este caso clínico confirma la importancia que tienen los animales que excretan Salmonella vía fecal y describe la relación epidemiológica-molecular de los aislamientos obtenidos de heces y alimento, lo que permitió esclarecer la fuente primaria de infección. Abstract in english Salmonella enterica serotypes are 99% responsible for salmonellosis in human and animals, especially Salmonella enterica serovar Albany that has been identified in chicken carcass representing a risk for human and animal health. Salmonella enterica serovar Albany was isolated from the feces of a mal [...] e ocelot (Leopardus pardalis), at the zoo in Culiacan, Sinaloa, Mexico, and from raw chicken (feline's diet). The pulsed-field gel electrophoresis pattern (PFGE) generated by Xba I enzyme was identical in both isolates, indicating that the source of infection was the raw chicken. Five months after having isolated the bacteria from the feces, a post mortem study was carried out on the feline. Macroscopically, severe hemorrhagic enterocolitis and renal fibrosis was observed and microscopically, there was evidence of severe mononuclear lymphocytic infiltration in the ileum, as well as necrosis of intestinal villi and crypts, besides severe multifocal interstitial nephritis and fibrosis in both kidneys. The invA gene was amplified from intestinal samples confirming an infection by Salmonella. The microbiologic, molecular and histopathology diagnoses suggest that death of the feline was caused by ingestion of raw chicken contaminated with Salmonella enterica serovar Albany. This clinical case highlights the importance of persistent fecal Salmonella shedding animals and describes the molecular epidemiological relationships of isolates from feces and food, which allowed to find the primary source of infection.

Gabriela, Silva-Hidalgo; Héctor Samuel, López-Moreno; Vianney F., Ortiz-Navarrete; Felipe, Juárez-Barranco; Martín, López-Valenzuela.

133

The Salmonella enterica Pan-genome  

DEFF Research Database (Denmark)

Salmonella enterica is divided into four subspecies containing a large number of different serovars, several of which are important zoonotic pathogens and some show a high degree of host specificity or host preference. We compare 45 sequenced S. enterica genomes that are publicly available (22 complete and 23 draft genome sequences). Of these, 35 were found to be of sufficiently good quality to allow a detailed analysis, along with two Escherichia coli strains (K-12 substr. DH10B and the avian pathogenic E. coli (APEC O1) strain). All genomes were subjected to standardized gene finding, and the core and pan-genome of Salmonella were estimated to be around 2,800 and 10,000 gene families, respectively. The constructed pan-genomic dendrograms suggest that gene content is often, but not uniformly correlated to serotype. Any given Salmonella strain has a large stable core, whilst there is an abundance of accessory genes, including the Salmonella pathogenicity islands (SPIs), transposable elements, phages, and plasmid DNA. We visualize conservation in the genomes in relation to chromosomal location and DNA structural features and find that variation in gene content is localized in a selection of variable genomic regions or islands. These include the SPIs but also encompass phage insertion sites and transposable elements. The islands were typically well conserved in several, but not all, isolates—a difference which may have implications in, e.g., host specificity.

Jacobsen, Annika; Hendriksen, Rene S.

2011-01-01

134

Detection of virulence-associated genes in Salmonella Enteritidis isolates from chicken in South of Brazil / Detecção de genes associados à virulência em cepas de Salmonella Enteritidis isoladas de frangos na região sul do Brasil  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese Salmonella spp. estão entre os principais agentes causadores de doenças transmitidas por alimentos, e o sorovar Salmonella Enteritidis é o mais frequentemente isolado no mundo. A virulência de Salmonella spp. e a sua interação com o hospedeiro são processos complexos que envolvem fatores de virulênc [...] ia para sobreviver às defesas do hospedeiro. O objetivo deste estudo foi detectar genes de virulência em cepas de S. Enteritidis isoladas a partir de fontes avícolas no sul do Brasil. Ensaios de PCR foram desenvolvidos para a detecção de nove genes (lpfA, agfA, sefA, invA, hilA, avrA, sopE, sivH e spvC) associados à virulência em oitenta e quatro amostras de S. Enteritidis. Os genes invA, hilA, sivH, sefA e avrA estavam presentes em 100% dos isolados; lpfA e sopE estavam presentes em 99%; agfA em 96%; e o gene spvC estava presente em 92%. Foi possível caracterizar os isolados em quatro perfis genéticos distintos (P1, P2, P3 e P4), sendo P1 positivo para todos os genes; P2 negativo apenas para spvC; P3 negativo para agfA e P4 negativo para lpfA, spvC e sopE. O perfil de maior frequência foi P1, presente em 88% dos isolados. Apesar de todos os isolados pertencerem ao mesmo sorovar, foi possível observar variações na presença de genes associados à virulência entre os mesmos. A caracterização dos mecanismos de virulência circulantes na população de Salmonella Enteritidis é importante para um maior entendimento da sua biologia e patogenicidade. A frequência destes genes e o estabelecimento de perfis genéticos podem ser utilizados para determinar os padrões de virulência dos isolados. Estes padrões, associados a estudos in vivo, podem auxiliar na elaboração de ferramentas que permitam predizer a capacidade de virulência das diferentes cepas. Abstract in english Salmonella spp. are considered the main agents of foodborne disease and Salmonella Enteritidis is one of the most frequently isolated serovars worldwide. The virulence of Salmonella spp. and their interaction with the host are complex processes involving virulence factors to overcome host defenses. [...] The purpose of this study was to detect virulence genes in S. Enteritidis isolates from poultry in the South of Brazil. PCR-based assays were developed in order to detect nine genes (lpfA, agfA, sefA, invA, hilA, avrA, sopE, sivH and spvC) associated with the virulence in eighty-four isolates of S. Enteritidis isolated from poultry. The invA, hilA, sivH, sefA and avrA genes were present in 100% of the isolates; lpfA and sopE were present in 99%; agfA was present in 96%; and the spvC gene was present in 92%. It was possible to characterize the isolates with four different genetic profiles (P1, P2, P3 and P4), as it follows: P1, positive for all genes; P2, negative only for spvC; P3, negative for agfA; and P4, negative for lpfA, spvC and sopE. The most prevalent profile was P1, which was present in 88% of the isolates. Although all isolates belong to the same serovar, it was possible to observe variations in the presence of these virulence-associated genes between different isolates. The characterization of the mechanisms of virulence circulating in the population of Salmonella Enteritidis is important for a better understanding of its biology and pathogenicity. The frequency of these genes and the establishment of genetic profiles can be used to determine patterns of virulence. These patterns, associated with in vivo studies, may help develop tools to predict the ability of virulence of different strains.

Karen A., Borges; Thales Q., Furian; Anderlise, Borsoi; Hamilton L.S., Moraes; Carlos T.P., Salle; Vladimir P., Nascimento.

1416-14-01

135

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)

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 ?leuO 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. PMID:22343301

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

136

Evaluation of two new chromogenic media for detection of Salmonella in stools.  

Science.gov (United States)

A clinical collaborative study was conducted to compare two new chromogenic agar media, Rambach agar and the Salmonella Detection and Identification Medium (SMID) (bioMérieux, France), with two conventional media, Salmonella-Shigella agar and Hektoen agar. Thirty-nine Salmonella strains involving 14 serotypes were isolated from 1,454 stool specimens. After enrichment in a selective broth, 100% sensitivity was obtained with each medium. The SMID and Rambach agars are considerably more specific than the conventional media. Although SMID agar detects all Salmonella serotypes, it is not as specific as Rambach agar, which requires a complementary test (C8 esterase test) to detect all serotypes. PMID:8050441

Monnery, I; Freydiere, A M; Baron, C; Rousset, A M; Tigaud, S; Boude-Chevalier, M; de Montclos, H; Gille, Y

1994-03-01

137

Salmonella newport and typhimurium colonization of fruit differs from leaves in various tomato cultivars.  

Science.gov (United States)

Several outbreaks of Salmonella enterica infections have been linked to tomatoes. One cost-effective way to complement on-farm preventive Good Agricultural Practices is to identify cultivars with inherent decreased susceptibility to Salmonella colonization. Fruit and leaves of 13 tomato cultivars with distinct phenotypes were screened to evaluate their susceptibility to Salmonella epiphytic colonization. Field-grown fruit or gnotobiotically grown seedling leaves were spot inoculated in replicate with either Salmonella Typhimurium LT2 or a tomato outbreak-associated strain of Salmonella Newport. Initial loads of the Salmonella inocula were 2.5 log CFU per fruit and 3.5 or 7.0 log CFU per seedling. Salmonella cells were retrieved and enumerated using direct plating after 24 h of incubation at room temperature for fruit and 72 h at 26°C during the day and 18°C at night for seedling leaves. Epiphytic colonization of fruit by S. enterica was cultivar-dependent and serotype-specific, but did not necessarily correlate with leaf colonization. Fruit of cultivar Heinz-1706 were the least colonized by Salmonella Newport, while the highest populations were retrieved from fruit of Nyagous. By contrast, seedling leaves supporting the lowest populations were Florida 91 VF and the highest were Virginia Sweets for Salmonella Newport. For Salmonella Typhimurium the lowest was Nyagous and the highest was Heinz-1706 and Moneymaker. The tomato outbreak strain of Salmonella Newport attained higher population densities on fruit than did Salmonella Typhimurium, suggesting better adaptation to tomato fruit colonization. Salmonella Newport populations were significantly lower on leaves, but not fruit of the near-isogenic line Movione, compared with the parent cultivar Moneymaker, suggesting the immunity conferring gene Pto could be responding to this outbreak strain. Susceptibility of tomato fruit to Salmonella colonization is highly variable and could be one criterion for cultivar selection for cultivation. PMID:25364916

Han, Sanghyun; Micallef, Shirley Ann

2014-11-01

138

Comparison of CHROMagar Salmonella medium and hektoen enteric agar for isolation of salmonellae from stool samples.  

Science.gov (United States)

CHROMagar Salmonella (CAS), a new chromogenic medium, was retrospectively compared to Hektoen enteric agar (HEA) with 501 Salmonella stock isolates and was then prospectively compared to HEA for the detection and presumptive identification of Salmonella spp. with 508 stool samples before and after enrichment. All stock cultures (100%), including cultures of H2S-negative isolates, yielded typical mauve colonies on CAS, while 497 (99%) isolates produced typical lactose-negative, black-centered colonies on HEA. Following overnight incubation at 37 degreesC, a total of 20 Salmonella strains were isolated from the 508 clinical samples. Sensitivities for primary plating and after enrichment were 95% (19 isolates) and 100% (20 isolates), respectively, for CAS and 80% (16 isolates) and 100% (20 isolates), respectively, for HEA. The specificity of CAS (88.9%) was significantly higher than that of HEA (78.5%; P medium can be recommended for use for primary plating when human stool samples are screened for Salmonella spp. PMID:9986847

Gaillot, O; di Camillo, P; Berche, P; Courcol, R; Savage, C

1999-03-01

139

Salmonella radicidation of poultry carcasses  

International Nuclear Information System (INIS)

This thesis reports investigations using gamma-radiation to decontaminate poultry carcasses. The application to foods of doses of ionizing radiation sufficient to reduce the number of viable specific non-sporeforming pathogenic microorganisms so that none is detectable in the treated food by any standard method is termed radicidation. The doses used in this study were at such a level that no undesirable or unfavourable side-effects occurred. The effects of these doses were studied on salmonellae and other microorganisms present in, or associated with poultry carcasses and in liquid and on solid culture media as well. Decimal reduction (D10) values were estimated. These represent the dose (kGy) required to achieve a reduction in initial colony count from N0 to 0.1 N0. Together with the estimation of the numbers of Salmonella present per carcass the data were used to predict the effect of an ionizing radiation treatment of poultry. Data on the effect of ionizing radiation on the total microflora of poultry carcasses were also collected. (Auth.)

140

Salmonella as an Innovative Therapeutic Antitumor Agent  

Directory of Open Access Journals (Sweden)

Full Text Available Lack of specificity of the therapeutic agent is a primary limitation in the treatment of a tumor. The use of preferentially replicating bacteria as therapeutic agents is an innovative approach to tumor treatment. This is based on the observation that certain obligate or facultative anaerobic bacteria are capable of multiplying selectively in tumors and inhibiting their growth. Bacteria have been employed as antitumor agents that are capable of preferentially amplifying within tumors and inhibiting their growth. Moreover, bacteria-derived factors have an immune-stimulation effect. Therefore, bacteria are able to transfer therapeutic genes into the tumor cells using their infective ability. Herein, we introduce the application of bacteria for tumor therapy and focus on Salmonella, which have been widely used for tumor therapy. Salmonella have mainly been applied as gene-delivery vectors, antitumor immune activators and tumor cell death inducers. This study will not only evaluate the therapeutic efficacy of Salmonella for the treatment of tumor but will also elucidate the mechanisms underlying the antitumor activities mediated by Salmonella, which involve host immune responses and cellular molecular responses.

Wen-Wei Chang

2014-08-01

 
 
 
 
141

Stably Integrated luxCDABE for Assessment of Salmonella Invasion Kinetics  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells tradition...

Flentie, Kelly N.; Qi, Min; Gammon, Seth T.; Razia, Yasmin; Lui, Felix; Marpegan, Luciano; Manglik, Aashish; Piwnica-worms, David; Mckinney, Jeffrey S.

2008-01-01

142

Comparison of Salmonella chromogenic medium with DCLS agar for isolation of Salmonella species from stool specimens.  

Science.gov (United States)

Salmonella chromogenic medium (SCM; Oxoid, Basingstoke, United Kingdom), a new selective chromogenic medium, was compared to DCLS agar (Oxoid) for the detection and presumptive identification of Salmonella species from stool samples. This medium contains two chromogenic substrates, Magenta-cap (5-bromo-6-chloro-3-indolylcaprylate), which is hydrolyzed by Salmonella species to give magenta colonies, and X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside), which is incorporated to visualize beta-D-galactosidase-producing organisms as blue colonies. Thus, non-Salmonella organisms appear blue or are not stained by any of the chromogens of the medium. A total of 500 stool samples were investigated by plating them directly and after selenite enrichment on DCLS agar and SCM. A total of 44 Salmonella-positive stool samples were detected. The sensitivities for direct plating and after enrichment were 22.7 and 81.8%, respectively, for DCLS agar, and for SCM these values were 34.1 and 100%, respectively. The specificities for direct plating and after enrichment were 82.5 and 72.8%, respectively, for DCLS agar and 98.5 and 95.8%, respectively, for SCM. According to these results, the sensitivities of SCM and DCLS agar were comparable on primary plating. However, the sensitivity of SCM was significantly higher after enrichment. In addition, the specificity of SCM was also significantly higher than that of DCLS agar both before and after enrichment. On the basis of these results, SCM can be recommended for the isolation of Salmonella species from stool samples in preference to DCLS agar. PMID:12843068

Cassar, Robert; Cuschieri, Paul

2003-07-01

143

 Salmonella multiphasic flagellar antigen  

Directory of Open Access Journals (Sweden)

Full Text Available  In the Salmonella antigenic pattern, more than one phase of flagellar antigen is observed. The phase of flagellar antigen depends of the gene which encodes the protein building the filament of flagella. The fliC gene encodes the 1st phase of flagellar antigen and the fljB gene encodes the 2nd phase of flagellar antigen. The third phase of flagellar antigen is encoded by one of the genes localized on the plasmid. Expression of the fljB gene (part of the hinfljBA operon is regulated by a mechanism of DNA fragment sequence inversion. The hin gene, which encodes Hin invertase, flanked by two regions – hixL and hixR – is inverted by Hin invertase together with Fis protein. This process turns on or turns off of the hinfljBA operon. When this operon is turned on, FljB protein is produced (structural protein of flagella filament, and also FljA protein, which is a transcriptional repressor of the fliC gene. This means that one Salmonella cell could have only one phase flagellar antigen – 1st or 2nd phase. Sometimes, due to mutation in one of the mentioned genes, naturally diphasic Salmonella strains have the ability to produce only one phase of flagellar antigen. Mostly monophasic Salmonella with an active fliC gene are observed. In recent years such a strain, Salmonella enterica with the antigenic formula 1,4,[5],12: i: -, is one of the most often isolated strains from human cases in many European countries.

Grzegorz Madajczak

2012-06-01

144

Salmonella – A Brief Summary  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Salmonellosis is the main cause of human bacterial gastroenteritis in most European countries. Infections with Salmonella is usually subclinical, whereas clinical cases show symptoms with a wide range of severity. Infection is most commonly associated with the consumption of meat, especially poultry or pork, and eggs and their products. Salmonella can enter the food chain at any point throughout its length. The principal reservoir of Salmonellae is the gastrointestinal tract of mammals and birds, but Salmonellae are able to survive and even multiply in many external environments. In Norway, Sweden and Finland cost effective prevention methods have been used for several years to prevent and control Salmonellea infections. In addition, competitive exclusion (CE and vaccination might be relevant as biological methods to prevent colonisation of bird intestines by enteropathogens, especially Salmonella. Antibiotic drug resistance has been a problem since the start of the antibiotic era. The cause for anxiety is that more and more bacteria are becoming resistant, often to a whole range of antibiotics. The debate on the use of antimicrobials in veterinary medicine and animal production dates back almost as long as the use itself. There is a clear evidence to show that antibacterial agents given to animals for growth promotion, prophylactic purposes or treatment induce a rise in the number of antibiotic resistant strains isolated from the animals. These bacteria may be transmitted to humans by several possible routes. There are thus strong arguments for preventive efforts which have to be directed towards identifying real critical control points (HACCP throughout the whole food chain, which starts from the farm and ends at the consumer's table.

Nurmi Esko

2002-03-01

145

Rapid detection of Salmonella in foods using a combination of SPRINT TM,MSRV TM and Salmonella Latex TestTM Detecção rápida de Salmonella em alimentos empregando uma combinação de SPRINT®, MSRV® e Salmonella Latex Test®  

Directory of Open Access Journals (Sweden)

Full Text Available A new procedure for rapid detection of Salmonella in foods, based on the combination of SPRINT TM, MSRV TM and Salmonella Latex TestTM, was evaluated. SPRINT TM is a system to reduce the preenrichment and selective enrichment steps to 24 hours. MSRV TM is a semi-solid selective media for detection of motile Salmonella. Salmonella Latex TestTM is a rapid latex agglutination test for Salmonella. Using the three systems in combination, the total time for detection of Salmonella in a food sample is 48h. Evaluations were performed in artificially contaminated ready-to-eat baby-foods and raw Brazilian sausages (lingüiça containing no added microorganisms. The BAM conventional culture procedure was used as reference method. The study with baby foods indicated that the new procedure had good sensitivity (89% and specificity (100%, without cross-reactions with Enterobacteriaceae. However, when applied to naturally contaminated foods, the performance was poor: chi square (x² = 5.062, ?> 0. 05 and Kappa-Cohen agreement (K = 0.171, p=0.089 indexes indicated that the differences between results given by the two procedures were significant and the correlation between them was low.Avaliou-se um novo procedimento para detecção rápida de Salmonella em alimentos, baseado na combinação entre SPRINT®, MSRV® e Salmonella Latex Test® . SPRINT® é um sistema para reduzir as etapas de pré-enriquecimento e enriquecimento seletivo para 24 h. MSRV® é um meio seletivo semi-sólido para detecção de salmonelas móveis. Salmonella Latex Test® é um teste rápido de aglutinação de látex. A combinação dos três sistemas permite que a detecção de Salmonella em alimentos possa ser feita em apenas 48 h. O procedimento foi avaliado em alimentos infantis prontos para consumo, experimentalmente contaminados com Salmonella exclusivamente e com uma mistura de Salmonella e várias espécies de Enterobacteriaceae e também em cem amostras de lingüiças de porco e de frango sem adição artificial de microrganismos. O método convencional de cultura foi empregado como método de referência. A avaliação em alimentos infantis indicou que o procedimento proposto apresentava boa sensibilidade (80% e especificidade (100%, sem reação cruzada com outras Enterobacteriaceae. Entretanto, quando aplicado a lingüiças, seu desempenho não foi adequado: os valores de x² (5,062, ? > 0,05 e do índice de concordância de Kappa (0,171, p=0,089 indicaram que as diferenças entre os resultados obtidos pelos dois métodos foram estatisticamente significantes e a correlação entre eles foi baixa.

Jane Maria Lafayette Neves Gelinski

2002-09-01

146

Mutations in Flk, FlgG, FlhA, and FlhE That Affect the Flagellar Type III Secretion Specificity Switch in Salmonella enterica?  

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Upon completion of the flagellar hook-basal body (HBB) structure, the flagellar type III secretion system switches from secreting rod/hook-type to filament-type substrates. The secretion specificity switch has been reported to occur prematurely (prior to HBB completion) in flk-null mutants (P. Aldridge, J. E. Karlinsey, E. Becker, F. F. Chevance, and K. T. Hughes, Mol. Microbiol. 60:630-643, 2006) and in distal rod gene gain-of-function mutants (flgG* mutants) that produce filamentous rod str...

Hirano, Takanori; Mizuno, Shino; Aizawa, Shin-ichi; Hughes, Kelly T.

2009-01-01

147

Application of bioinformatics on the detection of pathogens by Pcr  

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Salmonellas are the main responsible agent for the frequent food-borne gastrointestinal diseases. Their detection using classical methods are laborious and their results take a lot of time to be revealed. In this context, we tried to set up a revealing technique of the invA virulence gene, found in the majority of Salmonella species. After amplification with PCR using specific primers created and verified by bioinformatics programs, two couples of primers were set up and they appeared to be very specific and sensitive for the detection of invA gene. (Author)

148

Molecular Characterisation of Salmonella enterica Serovar Typhi Isolated from Typhoidial Humans  

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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.

Arunava Das

2012-09-01

149

Rapid Detection of Salmonella in Chicken Meat Using Immunomagnetic Separation, CHROMagar, ELISA and Real-time Polymerase Chain Reaction (RT-PCR  

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Full Text Available The main objective of this study was to standardize and compare rapid methods for the detection of Salmonella in meat samples using Immuno-Magnetic Separation (IMS followed by culturing in CHROMagar Plus media, Enzyme-Linked Immunosorbent Assay (ELISA and Real-Time Polymerase Chain Reaction (RT-PCR. Ten-fold dilutions of bacterial suspension (S. typhymurium, ATCC13311 were prepared from the original concentration of 1.6 x 106cfu/ml. Chicken wing samples of 25 g each negative for Salmonella were spiked with six different concentrations of bacteria ranging from 106 to 101. These samples were incubated in buffered peptone water for 4 h as pre-enrichment step and were tested repeatedly. The IMS technique involved the use of paramagnetic polystyrene microscopic beads coated with purified antibodies against Salmonella. The CHROMagar Plus media containing chromomeric substrate facilitated detection of Salmonella species from other flora. The Assurance EIA Salmonella Kit with polyclonal antibodies directed against Salmonella facilitated easy and rapid detection. In the RT-PCR primers targeting invA gene was used which amplified a 378 bp fragment. Comparing to conventional culture method (4 days, CHROMagar plate culture following IMS showed light mauve to mauve-colored colonies of Salmonella in 23 h with high sensitivity (99% at 1.6 cfu/ml. IMS-ELISA combination also showed high sensitivity (75% at 1.6 x 103 cfu/ml in 8 h and minimized cross-reactivity with many Enterobacteraceae. The combination of IMS with RT-PCR took less than 7 h and was even more sensitive (100% at 1.6 cfu/ml. Sensitivities of IMS-RT-PCR and IMS-CHROMagar were higher compared to IMS-ELISA. IMS-CHROMagar was easier to perform and detects only living Salmonella. These methods will be highly suitable for routine detection and may significantly assist the processing industry in avoiding costly recalls and the timely investigation of outbreaks.

K.K. Srivastava

2010-01-01

150

FECAL EXCRETION OF Salmonella Enteritidis IN BROILER LINES ROSS AND ISA LABEL EXCREÇÃO FECAL de Salmonella Enteritidis EM DUAS LINHAGENS DE FRANGOS DE CORTE  

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The invasive capacity and persistence of this pathogen, crop and ceca in apparently healthy birds of two broiler lines raised without growth promoter antibiotics in ration and originated from eggs inoculated eggshell and in allantoidal cavity with Salmonella Enteritidis. Histological and bacteriological exams from cecal and crop were performed with one, seven, 14 and 21 days of age after hatch in broilers of fast and slow growing rate. Bacterio-logical exams were performed fecal excretion with one, eigth, 22 and 35 days. The Salmonella Enteritidis invaded and colonizated the gastrointestinal tract of the two lines tested, but the the infection reduced with age, and was more persistant in Ross broilers. The results were different for two lines. The pathogen was excreted from just one chick of ISA Label at 22 days of age and four Ross chicks until 35 days of age. In order, Salmonella was detected in 87.5% (14/16 and 38,1% (5/16 of ceca; in 81.2% (13/16 and 12.5% (2/16 of crops; in fast and slow growing rate lines, respectively. In apparent healthy organs, excepted the crop, an inflammatory process with predominance of macrophage and lymphocytes. The slow growing rate line was effective to eliminate bacteria in the organism.

Key-words: Ceca, crop, fecal excretion, inflammation.

Avaliaram-se, neste estudo, a capacidade inva-siva, a persistência e a freqüência de excreção fecal da Salmonella Enteritidis em aves aparentemente saudáveis de duas linhagens de frango de corte, criadas sem antibióticos promotores de crescimento na ração e oriundas de ovos inoculados na casca ou na cavidade alantóide com Salmonella Enteritidis fagotipo 4. Realizaram-se exames bacteriológicos das excretas com um, oito, 22 e 35 dias, e histológicos e bacteriológicos do inglúvio e ceco, com um, sete, quatorze e 21 dias pós-eclosão em frangos de crescimento rápido e lento. Salmonella Enteritidis invadiu e colonizou o trato gastrintestinal das duas linhagens, mas a infecção declinou com a idade, sendo mais persistente na linhagem Ross. O patógeno foi excretado de uma única ave ISA Label até 22 dias de vida e em quatro aves da linhagem Ross até 35 dias. Constatou a Salmonella em ordem de colonização, em 87,5% (14/16 e 38,1% (5/16 dos cecos; em 81,2% (13/16 e 12,5% (2/16 dos inglúvios das linhagens Ross e ISA Label, respectivamente. Nos cecos aparentemente saudáveis, evidenciou-se um processo inflamatório com predominância de macrófagos e ou linfócitos, enquanto no inglúvio não se detectaram alterações microscópicas. A linhagem de ISA Label foi mais hábil em eliminar a bactéria do seu organismo.

Palavras-chaves: Ceco, excreção fecal, inflamação, inglúvio.

Adson Santa Cruz Oliveira

2007-12-01

151

Salmonella Questions and Answers  

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152

Salmonella in pigs  

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In recent years several incidents of feed-borne spread of Salmonella spp. have been documented in Swedish pig herds, including serotypes previously not associated with pigs. In this thesis two feed-associated serotypes (S Cubana and S Yoruba) were compared with two serotypes commonly detected in pigs (S Typhimurium and S Derby). The overall aim of the thesis was to increase knowledge about the feed-associated serotypes, with special focus on their infection dynamics in pigs. In 2003, a contam...

O?sterberg, Julia

2010-01-01

153

Evaluation of a new chromogenic medium for the isolation and presumptive identification of Salmonella species from stool specimens.  

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The performance of BBL CHROMagar Salmonella (Becton Dickinson, France), a new selective chromogenic medium for the isolation and presumptive identification of Salmonella spp., was evaluated. On this medium, which is a modification of CHROMagar Salmonella (CHROMagar Microbiology, France) with enhanced selectivity, the colonies of Salmonella are stained in mauve (rose-violet), while those of other organisms appear in blue-green or are not stained by any of the chromogens of the medium. The medium was evaluated with a total of 176 strains of Salmonella and other organisms, consisting of 18 reference strains and 158 clinical isolates. All Salmonella strains except subspecies IIIa and IIIb strains and Salmonella Gallinarum yielded typical mauve colonies. During the evaluation with 107 known positive and 332 unknown stool specimens in a clinical laboratory, a total of 115 and 105 Salmonella isolates were obtained on BBL CHROMagar Salmonella and Hektoen enteric agar, respectively. From the known positive stool specimens, 92 true positive cultures were obtained on BBL CHROMagar Salmonella and 89 on Hektoen enteric agar, yielding sensitivities of 86 and 83%, respectively. From the unknown stool specimens, a total of 27 Salmonella isolates were obtained, with 23 isolated from BBL CHROMagar Salmonella and 16 from Hektoen enteric agar by direct plating (sensitivity 85 and 59%, specificity 99 and 97%, respectively). Seroagglutination tests could be performed directly from BBL CHROMagar Salmonella. Compared to conventional isolation media, the time needed for confirmatory biochemical and serological tests was shortened by about 1 day when BBL CHROMagar Salmonella was used. On the basis of these results, the medium can be recommended for the primary isolation and presumptive identification of Salmonella spp. from clinical stool specimens. PMID:11681435

Eigner, U; Reissbrodt, R; Hammann, R; Fahr, A M

2001-08-01

154

Evanescent Wave Fiber Optic Biosensor for Salmonella Detection in Food  

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Full Text Available Salmonella enterica is a major food-borne pathogen of world-wide concern. Sensitive and rapid detection methods to assess product safety before retail distribution are highly desirable. Since Salmonella is most commonly associated with poultry products, an evanescent wave fiber-optic assay was developed to detect Salmonella in shell egg and chicken breast and data were compared with a time-resolved fluorescence (TRF assay. Anti-Salmonella polyclonal antibody was immobilized onto the surface of an optical fiber using biotin-avidin interactions to capture Salmonella. Alexa Fluor 647-conjugated antibody (MAb 2F-11 was used as the reporter. Detection occurred when an evanescent wave from a laser (635 nm excited the Alexa Fluor and the fluorescence was measured by a laser-spectrofluorometer at 710 nm. The biosensor was specific for Salmonella and the limit of detection was established to be 103 cfu/mL in pure culture and 104 cfu/mL with egg and chicken breast samples when spiked with 102 cfu/mL after 2–6 h of enrichment. The results indicate that the performance of the fiber-optic sensor is comparable to TRF, and can be completed in less than 8 h, providing an alternative to the current detection methods.

Arun K. Bhunia

2009-07-01

155

A novel chromogenic ester agar medium for detection of Salmonellae.  

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A novel agar medium, chromogenic Salmonella esterase (CSE) agar, for the differentiation of salmonellae is described. The agar contains peptones and nutrient extracts together with the following (grams per liter unless otherwise specified): 4-[2-(4-octanoyloxy-3, 5-dimethoxyphenyl)-vinyl]-quinolinium-1-(propan-3-yl carboxylic acid) bromide (SLPA-octanoate; bromide form), 0.3223; lactose, 14. 65; trisodium citrate dihydrate, 0.5; Tween 20, 3.0; ethyl 4-dimethylaminobenzoate, 0.035% (wt/vol), novobiocin, 70 mg liter-1. The key component of the medium is SLPA-octanoate, a newly synthesized ester formed from a C8 fatty acid and a phenolic chromophore. In CSE agar, the ester is hydrolyzed by Salmonella spp. to yield a brightly colored phenol which remains tightly bound within colonies. After 24 h of incubation at 37 or 42 degreesC, colonies of typical Salmonella spp. were burgundy colored on a transparent yellow background, whereas non-Salmonella spp. were white, cream, yellow or transparent. CSE agar was evaluated by using a panel of strains including a high proportion of Salmonella and non-Salmonella strains giving atypical reactions on other differential agars. The sensitivity (93.1%) of CSE agar for non-typhi salmonellae compared favorably with those of Rambach (82. 8%), xylose-lysine-deoxycholate (XLD; 91.4%), Hektoen-enteric (89.7%), and SM ID (91.4%) agars. The specificity (93.9%) was also comparable to those of other Salmonella media (SM ID agar, 95.9%; Rambach agar, 91.8%; XLD agar, 91.8%; Hektoen-enteric agar, 87.8%). Strains of Citrobacter freundii and Proteus spp. giving false-positive reactions with other media gave a negative color reaction on CSE agar. CSE agar enabled the detection of >30 Salmonella serotypes, including agona, anatum, enteritidis, hadar, heidelberg, infantis, montevideo, thompson, typhimurium, and virchow, which accounted for 91.8% of the salmonella isolates recorded by the Public Health Laboratory Service (Colindale, London, England) for 1997. PMID:9925620

Cooke, V M; Miles, R J; Price, R G; Richardson, A C

1999-02-01

156

Microencapsulated sorbic acid and nature-identical compounds reduced Salmonella Hadar and Salmonella Enteritidis colonization in experimentally infected chickens.  

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The reduction of Salmonella prevalence in broilers is a priority in European Union agricultural policies because treatment with antibiotics is forbidden by Regulation (EC) 2160/2003. Two trials were conducted to evaluate the efficacy of a microencapsulated blend of sorbic acid and nature-identical compounds (i.e., chemically synthesized botanicals; SAB) on the reduction of the cecal prevalence and contents of Salmonella enterica serovars Hadar and Enteritidis in experimentally infected chickens. In the first trial, 125 one-day-old Lohmann specific-pathogen-free chickens were assigned to one of the following treatments: negative control (not challenged and not treated), positive control (challenged and not treated), SAB0.3, SAB1, or SAB5 (challenged and treated with the microencapsulated blend included in the feed at 0.03, 0.1, or 0.5%, respectively). At 30 d of age, birds were infected with 10(6) cfu of Salmonella Hadar, and after 5, 10, or 20 d postinfection, 5, 10, and 10 birds per treatment, respectively, were killed and the cecal contents and liver and spleen samples were analyzed for Salmonella Hadar. In the second trial, 100 one-day-old Ross 708 chickens were assigned to 1 of 5 treatments: control (not treated), SAB0.3, SAB1, SAB2, or SAB5 (treated with the blend included in the feed at 0.03, 0.1, 0.2, or 0.5%, respectively). At 7 d of age, the birds were challenged with 10(5) cfu of Salmonella Enteritidis, and after 7, 14, or 24 d after challenge, 5, 5, and 10 birds per treatment, respectively, were killed and cecal contents were analyzed for Salmonella Enteritidis. Results showed that in the early stage of infection Salmonella prevalence was high in both studies, whereas at the end of the observation periods, the blends at 0.03, 0.1, and 0.5 in the challenge with Salmonella Hadar and at 0.2 and 0.5% in the challenge with Salmonella Enteritidis significantly reduced (by 2 log(10) cfu) the cecal content of Salmonella. This study showed that intestinal delivery of microencapsulated sorbic acid and nature-identical compounds can result in a 100-fold reduction of Salmonella at the intestinal level in broilers at slaughter age. PMID:21753203

Grilli, E; Tugnoli, B; Formigoni, A; Massi, P; Fantinati, P; Tosi, G; Piva, A

2011-08-01

157

Antibiotic susceptibility of Salmonella spp.: a comparison of two surveys with a 5 years interval  

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Full Text Available Salmonella infections are one of the major global public health problems. During the last decade, antibiotic resistance and multiresistance of Salmonella spp. have increased a great deal, especially in developing countries with an increased and indiscriminate use of antibiotics in the treatment of humans and animals. This study aims to investigate and compare antimicrobial susceptibility patterns of Salmonella during 2005 and 2010.A total of 186 Salmonella strain during 2005 and 140 Salmonella strain during 2010 were isolated from stool specimens using standard methods. The isolates were confirmed as Salmonella by using a battery of biochemical reactions. Specific antisera were used for serologic characterization of Salmonella strain. Antimicrobial susceptibility testing was performed by standard disk diffusion method using ampicillin, trimethoprim-sulfamethoxasole, ceftriaxon, chloramphenicol, nalidixic acid and ciprofloxacin.One hundred eighty (96.8% of 186 isolated Salmonella strains in 2005, and 133 (95% of 140 isolated Salmonella strain in 2010 are recognized as Salmonella Enteritidis. Sensitivity of Salmonella isolates during 2005 and 2010 were 91.9% and 92.9% to ampicillin, 95.7% and 97.1% to trimethoprim-sulfamethoxasole, 99.5% and 100% to chloramphenicol, 99.5% and 100% to ciprofloxacin, 98.9% and 97.1% to ceftriaxon, 73.1% and 95.7% to nalidixic acid, respectively.Sensitivity of Salmonella isolates to all tested antimicrobial agents except to ceftriaxon was been slightly improved over testing period. Resistance rate to ceftriaxon was higher in 2010 than in 2005, and this fact deserves attention. Significantly increase susceptibility rate to nalidixic acid was observed between the two surveys

Gordana Mijovi?

2012-02-01

158

Prevalence and Antibiotic Susceptibility Patterns of Bloodstream Salmonella Infections in a Tertiary Care Hospital, Dhaka  

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Full Text Available Blood stream Salmonella infections range from self-limiting infections to life-threatening sepsis causing significant mortality and morbidity worldwide and require rapid and aggressive anti-microbial treatment. The antibiotic resistance pattern of Salmonella is ever changing over time. Rational and correct use of antibiotics requires understanding of the pathogen and drug resistance patterns in a community. This study was conducted to determine the status of bloodstream Salmonella infection and their antibiotic susceptibility patterns in a tertiary care hospital at Dhaka, Bangladesh. Six hundred and fifty six blood samples collected from clinically diagnosed enteric fever patients from Dhaka Medical college Hospital, Dhaka during January 2012 to December 2012 were processed. Salmonella enterica serovar typhi and paratyphi isolates were identified by standard microbiological and biochemical procedures. Ninty four isolates of Salmonella typhi and 59 isolates of S. paratyphi were isolated. Average prevalence rate of Salmonella in blood was 24.8%. Young, neonates and elderly persons are more prone to Salmonella infection and males are more susceptible to Salmonella septicemia than females. Most of the isolates Salmonella spp. were Multi-drug Resistance (MDR and showed high resistance against cefixime, ceftriaxone, cefipime, ciprofloxacin, chloramphenicol and meropenem. Nalidixic acid was found to be effective against them. Specific antibiotic utilization strategies like antibiotic restriction, combination therapy and usage according to the standard antimicrobial susceptibility testing may help to decrease or prevent the emergence of resistance and incidence of blood stream infections.

Zahed Uddin Mahmood Khan

2013-01-01

159

Intestinal epithelial responses to Salmonella enterica serovar Enteritidis: effects on intestinal permeability and ion transport.  

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Salmonella infection of chickens that leads to potential human foodborne salmonellosis continues to be a major concern. Chickens serve as carriers but, in contrast to humans, rarely show any clinical signs including diarrhea. The present investigations aimed to elucidate whether the absence of diarrhea during acute Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) infection may be linked to specific changes in the electrophysiological properties of the chicken gut. Immediately after slaughter, intestinal pieces of the mid-jejunum and cecum of either commercial broiler or specific pathogen-free (SPF) chickens were mounted in Ussing chambers in 2 separate experimental series. Living Salmonella Enteritidis (3 × 10(9)) or Salmonella Enteritidis endotoxin (20 mg/L), or both, were added to the mucosal side for 1 h. In both experimental series, the Salmonella infection decreased the trans-epithelial ion conductance G(t) (P Salmonella endotoxin to the epithelial preparations from jejunum and cecum of SPF chicken had an effect similar to living bacteria. However, the endotoxin had no additional effect on the intestinal function in the presence of bacteria. The decreasing effect of Salmonella and or its endotoxin on G(t) could be partly reversed by serosal addition of histamine. To our knowledge, this is the first study to address the functional response of native intestinal epithelium of chicken to an in vitro Salmonella infection. For the first time, it can be reported that intestinal ion permeability of chicken decreases acutely by the presence of Salmonella. This type of response could counteract ion and fluid secretion and may thus, at least in part, explain why chickens do not develop overt diarrhea after Salmonella infection. PMID:23091155

Awad, W A; Aschenbach, J R; Khayal, B; Hess, C; Hess, M

2012-11-01

160

Preparation and structural analysis of oligosaccharide monophosphates obtained from the lipopolysaccharide of recombinant strains of Salmonella minnesota and Escherichia coli expressing the genus-specific epitope of Chlamydia lipopolysaccharide.  

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The lipopolysaccharide of the recombinant strain Salmonella minnesota r595-207 expressing the genus-specific epitope of Chlamydia lipopolysaccharide [Holst, O., Brade, L., Kosma, P. and Brade, H. (1991) J. Bacteriol, 173, 1862-1866] was sequentially de-O- and de-N-acylated by mild hydrazinolysis and treatment with 4 M KOH, respectively. The resulting mixture of compounds was separated by high-performance anion-exchange chromatography and gel-permeation chromatography, yielding four oligosaccharide phosphates two of which were readily identified by their 1H-NMR- and 13C-NMR spectra as alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha-D-Glcp N 1,4'-bisphosphate (tetrasaccharide bisphosphate; Kdo = 3-deoxy-D-manno-octulopyranosonic acid) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha-D- GlcpN 1,4'-bisphosphate (pentasaccharide bisphosphate) [Holst, O., Broer, W., Thomas-Oates, J.E., Mamat, U. and Brade, H. (1993) Eur. J. Biochem. 214, 703-710]. The structures of the other two compounds were established by chemical analysis, NMR spectroscopy, and fast-atom-bombardment mass spectrometry as alpha-Kdo- (2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha-D-GlcpN 1-phosphate (tetrasaccharide 1-phosphate) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha-D- GlcpN 1-phosphate (pentasaccharide 1-phosphate). alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6)-alpha/beta- D-GlcpN 4'-phosphate (tetrasaccharide 4'-phosphate) and alpha-Kdo-(2-8)-alpha-Kdo-(2-4)-alpha-Kdo-(2-6)-beta-D-GlcpN-(1-6) -alpha/beta-D-GlcpN 4'-phosphate (pentasaccharide 4'-phosphate) were prepared from the 1,4'-bisphosphates isolated from the recombinant strain Escherichia coli F515-207 by treatment with alkaline phosphatase and purification by high-performance anion-exchange chromatography and gel-permeation chromatography. Their structures were characterised by chemical analysis, NMR spectroscopy, and fast-bombardment mass spectrometry. PMID:7515346

Holst, O; Thomas-Oates, J E; Brade, H

1994-05-15

 
 
 
 
161

Characterization of an unusual Salmonella phage type DT7a and report of a foodborne outbreak of salmonellosis.  

Science.gov (United States)

Salmonella enterica subsp. enterica serovar 4,[5],12,i:- is a monophasic variant of Salmonella Typhimurium and its occurrence has markedly increased in several European countries in the last ten years. In June 2011, an outbreak of Salmonella 4,[5],12,i:- was reported among attendees of a wedding reception in the North-East of Italy. The source of this outbreak was identified as a cooked pork product served during the wedding reception. All Salmonella isolates from humans and the contaminated pork products were identified as Salmonella 4,[5],12,i:- and phage typed as DT7a. Afterwards, the farm where the pigs were raised was identified and sampled, and Salmonella Typhimurium was isolated from swine fecal samples. Despite the difference in serovar, these Salmonella Typhimurium isolates were also phage typed as DT7a. In the present study, Salmonella isolates from animals, humans and pork products during the outbreak investigation were subtyped by pulsed-field gel electrophoresis (PFGE), Multiple-Locus Variable number tandem repeats Analysis (MLVA), and resistance patterns, aiming to identify the most suitable subtyping methods to characterize isolates associated with this outbreak. In addition, a collection of epidemiologically unrelated strains of Salmonella 4,[5],12,i:- and Salmonella Typhimurium sharing the same phage type (DT7a) was similarly characterized in order to investigate their genetic relationship. This study provides a first snapshot of a rare Salmonella phage type, DT7a, associated with both Salmonella 4,[5],12,i:- and Salmonella Typhimurium. Moreover, the study demonstrated that in this specific context MLVA could be a reliable tool to support outbreak investigations as well as to assess the genetic relatedness among Salmonella isolates. PMID:25108760

Lettini, A A; Saccardin, C; Ramon, E; Longo, A; Cortini, E; Dalla Pozza, M C; Barco, L; Guerra, B; Luzzi, I; Ricci, A

2014-10-17

162

High-throughput Universal Probe Salmonella Serotyping (UPSS) by nanoPCR.  

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Salmonella enterica subsp. enterica serovar identification is of great importance with respect to outbreak monitoring and case verification. Therefore rapid, sensitive and cost efficient detection of Salmonella spp. is indispensable within microbiology labs. To amalgamate single tube isolate identification with Salmonella typing, we developed the high-throughput Universal Probe Salmonella Serotyping (UPSS) technique based on nano liter PCR. In comparison to the classical approach, where O- and H-antisera are applied, the UPSS relies on specific gene content amplification of Salmonella spp. by a universal TaqMan assay for all markers and identification of the specific amplicon pattern. To enable high-throughput technology we employed a chip format containing 1024 wells loaded by an automated liquid-handling system which allowed us to perform TaqMan PCR reactions in volumes of 100nL per well. Herein we present proof of principle of the UPSS method by the use of a test panel of 100 previously serotyped Salmonella isolates to successfully verify the usability, accuracy and feasibility of the newly developed UPSS approach. We found that the methodology of the UPSS technology is capable of unequivocally identifying 30 Salmonella serotypes on a single chip within 3 hours but can be highly parallelized by the use of multiple PCR machines. Therefore the UPSS method offers a robust and straightforward molecular alternative for Salmonella detection and typing that saves expensive chemistry and can be easily automated. PMID:20869995

Mertes, Florian; Biens, Katja; Lehrach, Hans; Wagner, Martin; Dahl, Andreas

2010-11-01

163

Induction of seroconversion and persistence of Salmonella Typhimurium in pigs are strain dependent.  

Science.gov (United States)

Foodborne salmonellosis is one of the most important bacterial zoonotic diseases worldwide. Salmonella Typhimurium is the serovar most frequently isolated from persistently infected slaughter pigs in Europe. Salmonella Typhimurium pathogenesis is host species specific. In addition, differences in in vitro behaviour of Salmonella Typhimurium strains have also been described, which may be reflected by a different course of infection within a host species. We compared the course of a Salmonella Typhimurium infection in pigs, using two Salmonella Typhimurium strains that were able to interfere with MHC II expression on porcine macrophages to a different extent in vitro. After experimental inoculation, blood and faecal samples from all pigs were collected at regular time points. At 40 days post inoculation (pi), animals were euthanized and tissue samples were bacteriologically analysed. The proportion of serologically positive piglets at 33 days pi was significantly higher in pigs that were inoculated with the strain that did not downregulate MHC II expression in vitro. Furthermore, this strain was less frequently shed and isolated in lower numbers from tonsils and ileocaecal lymph nodes than the strain that was able to markedly downregulate MHC II expression in vitro. We thus found that the delayed onset of seroconversion after oral inoculation of piglets with a particular Salmonella Typhimurium strain coincided with higher faecal shedding and increased persistence. Strain specific differences in Salmonella pathogenesis might thus have repercussions on the serological detection of Salmonella Typhimurium infections in pigs. PMID:23669459

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

2013-09-01

164

Specific immunoglobulin A-secreting cells in peripheral blood of humans following oral immunization with a bivalent Salmonella typhi-Shigella sonnei vaccine or infection by pathogenic S. sonnei.  

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The ability of bivalent Salmonella typhi-Shigella sonnei vaccine strain 5076-1C to stimulate an intestinal immunoglobulin A response in humans was evaluated by detecting gut-derived, trafficking antibody-secreting cells (ASC) in peripheral blood. Following vaccination, an immunoglobulin A-ASC response to O antigens of S. typhi and S. sonnei was observed in 10 of 13 and 13 of 13 vaccine recipients, respectively. Experimental challenge with pathogenic S. sonnei stimulated an ASC response to the...

Verg, L.; Herrington, D. A.; Murphy, J. R.; Wasserman, S. S.; Formal, S. B.; Levine, M. M.

1990-01-01

165

Comparison of Four Chromogenic Media and Hektoen Agar for Detection and Presumptive Identification of Salmonella Strains in Human Stools  

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Several chromogenic media have been developed to enhance the specificity of Salmonella detection. We compared the performance of four commercial chromogenic media—namely, ABC medium (Lab M. Ltd., Bury, United Kingdom), COMPASS Salmonella agar (Biokar Diagnostics, Beauvais, France), CHROMagar Salmonella agar (CHROMagar Company, Paris, France), and SM ID agar (bioMerieux, Marcy l'Etoile, France)—with conventional Hektoen medium. Nine hundred sixteen stool samples from inpatients at three ho...

Perez, J. M.; Cavalli, P.; Roure, C.; Renac, R.; Gille, Y.; Freydiere, A. M.

2003-01-01

166

Development of Rapid Detection and Genetic Characterization of Salmonella in Poultry Breeder Feeds  

Directory of Open Access Journals (Sweden)

Full Text Available Salmonella is a leading cause of foodborne illness in the United States, with poultry and poultry products being a primary source of infection to humans. Poultry may carry some Salmonella serovars without any signs or symptoms of disease and without causing any adverse effects to the health of the bird. Salmonella may be introduced to a flock by multiple environmental sources, but poultry feed is suspected to be a leading source. Detecting Salmonella in feed can be challenging because low levels of the bacteria may not be recovered using traditional culturing techniques. Numerous detection methodologies have been examined over the years for quantifying Salmonella in feeds and many have proven to be effective for Salmonella isolation and detection in a variety of feeds. However, given the potential need for increased detection sensitivity, molecular detection technologies may the best candidate for developing rapid sensitive methods for identifying small numbers of Salmonella in the background of large volumes of feed. Several studies have been done using polymerase chain reaction (PCR assays and commercial kits to detect Salmonella spp. in a wide variety of feed sources. In addition, DNA array technology has recently been utilized to track the dissemination of a specific Salmonella serotype in feed mills. This review will discuss the processing of feeds and potential points in the process that may introduce Salmonella contamination to the feed. Detection methods currently used and the need for advances in these methods also will be discussed. Finally, implementation of rapid detection for optimizing control methods to prevent and remove any Salmonella contamination of feeds will be considered.

Steven C. Ricke

2009-07-01

167

Occurrence, growth, and suppression of salmonellae in composted sewage sludge.  

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Composted sewage sludge may be used to improve soil quality, but there remains some doubt concerning the microbiological safety of the product. Sewage sludge composts from 30 municipalities were sampled, and four samples (12%) contained salmonellae (two contained fewer than 0.3/g, and the other two had 21/g and 1.7 X 10(4)/g). All 30 composts were inoculated with salmonellae; the populations decreased at a specific death rate of about 0.15 h-1 over 24 h at 36 degrees C. In irradiation-sterili...

Hussong, D.; Burge, W. D.; Enkiri, N. K.

1985-01-01

168

Salmonella in Sheep in Iceland  

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Full Text Available In 1995 several outbreaks of food poisoning in humans occurred in Iceland, that were traced to salmonella contamination of singed sheep heads. This prompted us to study the prevalence of salmonella infection in sheep and to trace where and how infection might have occurred. Faecal, intestinal contents and tonsillar samples were collected in the spring and autumn from sheep on 50 farms in the southwestern part of the country, where salmonellosis had been detected and from 5 farms in the northwestern part of the country. All faecal samples from the southwest were negative, whereas samples from 3 farms obtained in the autumn in the northwest were positive. Tonsillae taken in the autumn were positive in sheep from 3 farms in the southwest and 2 in the northwest. Our results show that salmonella infection is rare in Icelandic sheep but healthy carriers may harbour the bacteria in tonsillae. Salmonella was not detected in drainage from slaughterhouses nor in singed sheep heads.

Gunnarsson E

2002-03-01

169

Salmonella Control Programs in Denmark  

DEFF Research Database (Denmark)

We describe Salmonella control programs of broiler chickens, layer hens, and pigs in Denmark. Major reductions in the incidence of foodborne human salmonellosis have occurred by integrated control of farms and food processing plants. Disease control has been achieved by monitoring the herds and flocks, eliminating infected animals, and diversifying animals (animals and products are processed differently depending on Salmonella status) and animal food products according to the determined risk. In 2001, the Danish society saved U.S.$25.5 million by controlling Salmonella. The total annual Salmonella control costs in year 2001 were U.S.$14.1 million (U.S.$0.075/kg of pork and U.S.$0.02/kg of broiler or egg). These costs are paid almost exclusively by the industry. The control principles described are applicable to most industrialized countries with modern intensive farming systems.

Wegener, Henrik Caspar; Hald, Tine

2003-01-01

170

Comparison of Six Culture Methods for Salmonella Isolation from Poultry Fecal Samples  

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Full Text Available Background and Objective: Salmonellosis is one of the most important food-borne bacterial zoonotic diseases worldwide, and poultry and its products are the major sources for salmonella transmission to human. Isolation of Salmonella enterica from poultry needs bacteriologic enrichment and selected cultures of fecal samples. In this study, different culture methods for the isolation of salmonella from fecal samples were compared. Material and Methods: Forty- five positive samples from infected farms and 45 negative samples from normal farms were processed using enrichment media including tetrathionate broth, selenite cistine and Rappaport-Vassiliadis. Then the samples were incubated in selective cultures, and after 24 h, their results were compared with standard method. Results: Specificity of all methods for salmonella isolation was 100%, and salmonella was not isolated from the negative samples. The highest susceptibility was related to the method in which the sample first in Selenite cistine and later in Rappaport-Vassiliadis was enriched (100%. Enrichment in Rappaport-Vassiliadis could isolate 41 salmonella from 45 positive samples (91% while the result of enrichment in tetrathionate was 6 isolates (13.3%. Conclusion: This study shows that enrichment in selenite cistine and then in Rappaport-Vassiliadis is currently the best method for isolating salmonella from fecal samples of poultry. Key words: Salmonella; Bacteriologic Culture; Diagnosis; Isolation; Enrichment; Poultry

Morshed, R. (PhD

2014-06-01

171

Epidemiological Characterization of Avian Salmonella enterica Serovar Infections in India  

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Full Text Available The study was undertaken to elucidate the recent epidemiological status of salmonellosis in India. A total of 23 Salmonella isolates were recovered from different disease outbreaks in different geographical locations. The phenotypic analysis by serotyping, transmission electron microscopy and antibiogram profiles were able to classify the Salmonella isolates based on different serovars. The S. Gallinarum isolates were also classified into different strains based on the resistance pattern to different antimicrobials used. The plasmid profiles of the Salmonella isolates were also found to be serovar specific and the S. Gallinarum isolates were subdivided into different strains based on the geographical origin. There was a positive correlation between the antibiotic resistance pattern and the presence of plasmid within different serovars.

B. Prakash

2005-01-01

172

Effect of Low Dose of Fumonisins on Pig Health: Immune Status, Intestinal Microbiota and Sensitivity to Salmonella  

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Full Text Available The objective of this study was to measure the effects of chronic exposure to fumonisins via the ingestion of feed containing naturally contaminated corn in growing pigs infected or not with Salmonella spp. This exposure to a moderate dietary concentration of fumonisins (11.8 ppm was sufficient to induce a biological effect in pigs (Sa/So ratio, but no mortality or pathology was observed over 63 days of exposure. No mortality or related clinical signs, even in cases of inoculation with Salmonella (5 × 104 CFU, were observed either. Fumonisins, at these concentrations, did not affect the ability of lymphocytes to proliferate in the presence of mitogens, but after seven days post-inoculation they led to inhibition of the ability of specific Salmonella lymphocytes to proliferate following exposure to a specific Salmonella antigen. However, the ingestion of fumonisins had no impact on Salmonella translocation or seroconversion in inoculated pigs. The inoculation of Salmonella did not affect faecal microbiota profiles, but exposure to moderate concentrations of fumonisins transiently affected the digestive microbiota balance. In cases of co-infection with fumonisins and Salmonella, the microbiota profiles were rapidly and clearly modified as early as 48 h post-Salmonella inoculation. Therefore under these experimental conditions, exposure to an average concentration of fumonisins in naturally contaminated feed had no effect on pig health but did affect the digestive microbiota balance, with Salmonella exposure amplifying this phenomenon.

Philippe Fravalo

2013-04-01

173

Effect of low dose of fumonisins on pig health: immune status, intestinal microbiota and sensitivity to Salmonella.  

Science.gov (United States)

The objective of this study was to measure the effects of chronic exposure to fumonisins via the ingestion of feed containing naturally contaminated corn in growing pigs infected or not with Salmonella spp. This exposure to a moderate dietary concentration of fumonisins (11.8 ppm) was sufficient to induce a biological effect in pigs (Sa/So ratio), but no mortality or pathology was observed over 63 days of exposure. No mortality or related clinical signs, even in cases of inoculation with Salmonella (5 × 10? CFU), were observed either. Fumonisins, at these concentrations, did not affect the ability of lymphocytes to proliferate in the presence of mitogens, but after seven days post-inoculation they led to inhibition of the ability of specific Salmonella lymphocytes to proliferate following exposure to a specific Salmonella antigen. However, the ingestion of fumonisins had no impact on Salmonella translocation or seroconversion in inoculated pigs. The inoculation of Salmonella did not affect faecal microbiota profiles, but exposure to moderate concentrations of fumonisins transiently affected the digestive microbiota balance. In cases of co-infection with fumonisins and Salmonella, the microbiota profiles were rapidly and clearly modified as early as 48 h post-Salmonella inoculation. Therefore under these experimental conditions, exposure to an average concentration of fumonisins in naturally contaminated feed had no effect on pig health but did affect the digestive microbiota balance, with Salmonella exposure amplifying this phenomenon. PMID:23612754

Burel, Christine; Tanguy, Mael; Guerre, Philippe; Boilletot, Eric; Cariolet, Roland; Queguiner, Marilyne; Postollec, Gilbert; Pinton, Philippe; Salvat, Gilles; Oswald, Isabelle P; Fravalo, Philippe

2013-04-01

174

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

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

Jorge Hernández

2012-08-01

175

Development of an ultra rapid and simple multiplex polymerase chain reaction technique for detection of Salmonella typhi.  

Directory of Open Access Journals (Sweden)

These data indicate that the specificity and sensitivity of the PCR and the multiplex PCR make them potentially valuable tools for diagnosis of Salmonella typhi bacteria and that they may be used for the identification of Salmonella enteritidis responsible for sporadic enteritis cases.

Karami Ali

2006-08-01

176

Evaluation and implementation of a chromogenic agar medium for salmonella detection in stool in routine laboratory diagnostics.  

Science.gov (United States)

We evaluated which chromogenic agar medium for Salmonella detection in stool would be most sensitive and specific in our culture protocol. The use of BBL CHROMagar Salmonella chromogenic medium combined with xylose-lysine-deoxycholate agar yielded a sensitivity of 100% and also reduced workload and costs. PMID:19091816

van Dijk, Saskia; Bruins, Marjan J; Ruijs, Gijs J H M

2009-02-01

177

Evaluation and Implementation of a Chromogenic Agar Medium for Salmonella Detection in Stool in Routine Laboratory Diagnostics?  

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We evaluated which chromogenic agar medium for Salmonella detection in stool would be most sensitive and specific in our culture protocol. The use of BBL CHROMagar Salmonella chromogenic medium combined with xylose-lysine-deoxycholate agar yielded a sensitivity of 100% and also reduced workload and costs.

Dijk, Saskia; Bruins, Marjan J.; Ruijs, Gijs J. H. M.

2009-01-01

178

[Spondylodiscitis due to Salmonella enteritica serotype Typhi].  

Science.gov (United States)

We reported a case of lombar spondylodiscitis caused by Salmonella enteritica serotype Typhi in an immunocompetent patient. Salmonella is a rare causative agent of spondylodiscitis. Early bacteriological diagnosis is essential to avoid longterm sequelae. PMID:16230288

Zebouh, M; Loïez, C; Marceau, L; Vieillard, M H; Izard, D; Courcol, R J

2005-01-01

179

Health Tip: Avoiding Salmonella from Your Pet  

Science.gov (United States)

... this page, please enable JavaScript. Health Tip: Avoiding Salmonella From Your Pet Wash hands after handling food ... can be contaminated with nasty germs such as salmonella . The U.S. Centers for Disease Control and Prevention ...

180

Salmonella enteritidis deposition in eggs after experimental infection of laying hens with different oral doses.  

Science.gov (United States)

The continuing attribution of human Salmonella Enteritidis infections to internally contaminated eggs has necessitated the commitment of substantial public and private resources to Salmonella Enteritidis testing and control programs in commercial laying flocks. Cost-effective risk-reduction requires a detailed and comprehensive understanding of how Salmonella Enteritidis infections in hens result in deposition of the pathogen inside eggs. The present study sought to resolve some incompletely defined aspects of the relationship between Salmonella Enteritidis oral-exposure dose levels in experimentally infected laying hens and the frequency and location of subsequent egg contamination. In two trials, groups of specific-pathogen-free hens were experimentally inoculated with oral doses of 10(4), 10(6), or 10(8) CFU of a phage type 4 Salmonella Enteritidis strain. Eggs were collected 5 to 23 days postinoculation, and the yolk and albumen of each egg were cultured separately to detect Salmonella Enteritidis contamination. Larger oral doses of Salmonella Enteritidis administered to hens were associated with significant increases in the frequencies of both yolk and albumen contamination. Moreover, Salmonella Enteritidis was found in the albumen of a far-higher proportion of contaminated eggs from hens given the largest dose than from the other two groups. Salmonella Enteritidis contamination was detected in 0.7% of yolk and 0.2% of albumen samples after inoculation of hens with 10(4) CFU, 4.0% of yolk and 1.7% of albumen samples after inoculation with 10(6) CFU, and 6.5% of yolk and 10.8% of albumen samples after inoculation with 10(8) CFU. These results demonstrate that oral-exposure doses of Salmonella Enteritidis for laying hens can significantly affect both the frequency and location of deposition of this pathogen inside eggs. PMID:23317864

Gast, Richard K; Guraya, Rupa; Guard, Jean

2013-01-01

 
 
 
 
181

Salmonella: Clinical Importance and Evolution of Nomenclature  

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Salmonella is an important pathogen for both humans andanimals. Although the organism has been intensively studiedduring the last century, much remains to be learned about thispathogen. The complicated nomenclature system of Salmonellahas long been a subject of discussion. In 2005, “Salmonellaenterica” finally gained official approval as the type species ofthe genus Salmonella. The genus Salmonella also contains thespecies “Salmonella bongori” in addition to a new species,“Salmonell...

Cheng-Hsun Chiu; Lin-Hui Su

2007-01-01

182

Splenic abscess due to Salmonella enteritidis  

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Splenic abscess is a very rare complication of non-typhoidal Salmonella infections. We report a case of splenic abscess caused by Salmonella enteritidis. The patient is a 63-year-old woman with diabetes mellitus and underwent splenectomy. This case suggests that the patients with comorbities are at increased risk for invasive infections in non-typhoidal Salmonella infections.

Hatice Çabadak; Kerem Karaman; Süha ?en; Ay?e Erbay; Yasemin Tezer Tekçe

2012-01-01

183

Analysis of the FoodNet case-control study of sporadic Salmonella serotype Enteritidis infections using persons infected with other Salmonella serotypes as the comparison group.  

Science.gov (United States)

Use of well persons as the comparison group for laboratory-confirmed cases of sporadic salmonellosis may introduce ascertainment bias into case-control studies. Data from the 1996-1997 FoodNet case-control study of laboratory-confirmed Salmonella serogroups B and D infection were used to estimate the effect of specific behaviours and foods on infection with Salmonella serotype Enteritidis (SE). Persons with laboratory-confirmed Salmonella of other serotypes acted as the comparison group. The analysis included 173 SE cases and 268 non-SE controls. SE was associated with international travel, consumption of chicken prepared outside the home, and consumption of undercooked eggs prepared outside the home in the 5 days prior to diarrhoea onset. SE phage type 4 was associated with international travel and consumption of undercooked eggs prepared outside the home. The use of ill controls can be a useful tool in identifying risk factors for sporadic cases of Salmonella. PMID:18611288

Voetsch, A C; Poole, C; Hedberg, C W; Hoekstra, R M; Ryder, R W; Weber, D J; Angulo, F J

2009-03-01

184

Elisa indireto na detecção de Salmonella spp. em lingüiça suína / An indirect Elisa for detection of salmonella spp. in swine sausage  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese Um teste ELISA, baseado em um anticorpo monoclonal (MAb) específico para uma proteína de membrana externa de Salmonella enterica serovar Enteritidis foi comparado com o método de cultivo tradicional na detecção de Salmonella spp. em 110 amostras de lingüiça suína frescal. A prevalência do patógeno n [...] as amostras foi de 11,82% de acordo com o cultivo tradicional. O teste ELISA revelou sensibilidade, especificidade, valores preditivos positivos e negativos de 100%, 98%, 87% e 100%, respectivamente. Tais achados indicam que, comparado ao sistema tradicional, o teste imunológico foi bastante eficaz na determinação de Salmonella spp. em amostras de lingüiças naturalmente contaminadas. Abstract in english The performance of an ELISA test based on a monoclonal antibody (MAb) specific to Salmonella enterica serovar Enteritidis was compared with standard culture method for detection of Salmonella spp. in 110 samples of swine fresh sausages. The prevalence was of 11.82%, according the standard method. Co [...] mparison of ELISA and the culture method revealed the sensitivity, specificity, and positive and negative predictive values of 100%, 98%, 87% and 100%, respectively. Results indicate that compared with standard culture method the ELISA test was effective in detection of Salmonella in swine sausages naturally contaminated.

Andrea Pinto, Loguercio; José Antonio Guimarães, Aleixo; Agueda Castagna de, Vargas; Mateus Matiuzzi da, Costa.

185

Sources and fate of Salmonella and fecal indicator bacteria in an urban creek.  

Science.gov (United States)

This research aimed to understand the sources and fate of Salmonella and fecal bacteria in urban surface waters. An urban creek (San Pedro Creek, California, USA) that had unusually high levels of Salmonella and fecal bacteria relative to other nearby waterbodies was chosen as a model field site. State of the art microbiological methods were used in concert with modeling to investigate Salmonella and fecal bacteria sources, and determine field-relevant dark inactivation and photoinactivation rates. Three along-creek surveys that spanned reaches adjacent to both urban and forested land covers were conducted to measure Salmonella, enterococci, Escherichia coli, and horse- and human-specific Bacteroidales. Salmonella were detected adjacent to and downstream of urban land cover, but not adjacent to forested land cover. No human or horse-specific Bacteroidales fecal markers were detected implicating other urban animal sources of bacteria. Two locations along the creek where Salmonella was consistently detected were sampled hourly for 25 hours and a mass-balance model was applied to determine field-relevant light and dark inactivation rates for Salmonella, enterococci, and E. coli. Sunlight inactivation did not appear to be important in modulating concentrations of Salmonella, but was important in modulating both enterococci and E. coli concentrations. Dark inactivation was important for all three organisms. This is the first study to quantitatively examine the fate of Salmonella within an urban surface water. Although the work is carried out at a single site, the methodologies are extendable to source tracking in other waterbodies. Additionally, the rate constants determined through the modeling will be useful for modeling these organisms in other surface waters, and represent useful benchmarks for comparison to laboratory-derived inactivation rates. PMID:21687857

Sassoubre, Lauren M; Walters, Sarah P; Russell, Todd L; Boehm, Alexandria B

2011-08-01

186

Research and identification of pathogenic bacteria 'Salmonella and Listeria' in food  

International Nuclear Information System (INIS)

The sums propose to evaluate the bacterial contamination of certain food taken randomly by two pathogenic bacteria (Salmonella and Listeria) considering the evolution of the diseases of food oignon. For that 78 food samples of different origins were analysed. 2 stocks of the Listeria kind and 3 stocks of the salmonella kind were insulated and identified by biochemical and molecular tests. The pathogenic isolates were identified by coloration gram, test catalase, insulation on specific culture media and Api (20 E for Salmonella and Api listeria. At the end, the PCR were realized to amplify the gene iap which codes for the protein p60 at listeria as well as a sequence clonee randomly specific of Salmonella.

187

Salmonella detection using 16S ribosomal DNA/RNA probe-gold nanoparticles and lateral flow immunoassay.  

Science.gov (United States)

An ultrasensitive, simple, and fast lateral flow immunoassay for Salmonella detection using gold nanoparticles conjugated with a DNA probe, which is complementary to the 16S ribosomal RNA and DNA of Salmonella, has been developed. The detection limit is 5 fmol for the synthetic single-stranded DNA. For the Salmonella cultured samples, the nucleic acids from 10(7) bacteria were rapidly detected in 30 min. After silver enhancement, the detection limit was as low as 10(4) cells which is lower than 10(5) bacteria cells, the human infective dose of food-borne Salmonella. Furthermore, the probes used in this study are specific to Salmonella compared to several other Enterobacteriaceae. This approach would be a useful tool for microbial detection regarding food safety or clinical diagnosis. It is also suitable for large-scale screening in developing countries because it is low-cost, sensitive, specific and convenient. PMID:23870991

Liu, Cheng-Che; Yeung, Chun-Yan; Chen, Po-Hao; Yeh, Ming-Kung; Hou, Shao-Yi

2013-12-01

188

Comparison of 2 culture methods and PCR assays for Salmonella detection in poultry feces.  

Science.gov (United States)

The present work compared 2 culture methods and the combinations of pre-enrichment and enrichment culture methods with PCR assays [buffered peptone water-PCR and tetrathionate-PCR or modified semisolid Rappaport-Vassiliadis (MSRV)-PCR] for motile and nonmotile Salmonella strain detection using artificially contaminated poultry feces. The specificity and positive predictive values were equal to one in both culture methods. Specificity and positive predictive values, accuracy, sensitivity, and negative predictive values were higher for motile than nonmotile Salmonella strains in culture methods. Only Salmonella enterica serovar Gallinarum was detected by the MSRV method with low accuracy, sensitivity, and negative predictive value. The detection level of motile strains was 2 ×10(0) to 22 × 10(2) cfu per 25 g for these methods, whereas it was 6.9 × 10(2) cfu per 25 g in culture methods for Salmonella Gallinarum. Extending the incubation time of the enrichment medium to 6 d in the TT method did not improve the isolation rates. In general, all selective plating media did not show any statistical differences in the parameters of performance studied. On the other hand, accuracy and sensitivity values were higher in MSRV-PCR and tetrathionate-PCR methods than in the buffered peptone water-PCR method. Specificity and positive predictive values were equal to one in most of the cases. In terms of detection limits, motile Salmonella strains were recovered from 5 × 10(0) cfu per 25 g in MSRV-PCR and tetrathionate-PCR methods, whereas the detection limit was better for nonmotile Salmonella in MSRV-PCR methods than in the tetrathionate-PCR method. Kappa coefficients showed that there was a very good agreement between tetrathionate and MSRV methods for motile Salmonella strains, whereas these methods did not show any concordance for nonmotile Salmonella strains. When buffered peptone water-PCR was compared with both tetrathionate-PCR and MSRV-PCR, agreement was poor for motile Salmonella strains and slight to fair for nonmotile Salmonella strains. The difference in isolation rate obtained with the methods used for motile and nonmotile Salmonella strains must be taken into account when a poultry fecal sample is considered negative for the presence of Salmonella. PMID:22334736

Soria, M C; Soria, M A; Bueno, D J

2012-03-01

189

Antimicrobial resistance in typhoidal salmonellae.  

Science.gov (United States)

Infections with Salmonella are an important public health problem worldwide. On a global scale, it has been appraised that Salmonella is responsible for an estimated 3 billion human infections each year. The World Health Organization (WHO) has estimated that annually typhoid fever accounts for 21.7 million illnesses (217,000 deaths) and paratyphoid fever accounts for 5.4 million of these cases. Infants, children, and adolescents in south-central and South-eastern Asia experience the greatest burden of illness. In cases of enteric fever, including infections with S. Typhi and S. Paratyphi A and B, it is often necessary to commence treatment before the results of laboratory sensitivity tests are available. Hence, it is important to be aware of options and possible problems before beginning treatment. Ciprofloxacin has become the first-line drug of choice since the widespread emergence and spread of strains resistant to chloramphenicol, ampicillin, and trimethoprim. There is increase in the occurrence of strains resistant to ciprofloxacin. Reports of typhoidal salmonellae with increasing minimum inhibitory concentration (MIC) and resistance to newer quinolones raise the fear of potential treatment failures and necessitate the need for new, alternative antimicrobials. Extended-spectrum cephalosporins and azithromycin are the options available for the treatment of enteric fever. The emergence of broad spectrum ?-lactamases in typhoidal salmonellae constitutes a new challenge. Already there are rare reports of azithromycin resistance in typhoidal salmonellae leading to treatment failure. This review is based on published research from our centre and literature from elsewhere in the world. This brief review tries to summarize the history and recent trends in antimicrobial resistance in typhoidal salmonellae. PMID:21860101

Harish, B N; Menezes, G A

2011-01-01

190

Nutritional strategies to combat Salmonella in mono-gastric food animal production.  

Science.gov (United States)

Nutritional strategies to minimize Salmonella in food animal production are one of the key components in producing safer food. The current European approach is to use a farm-to-fork strategy, where each sector must implement measures to minimize and reduce Salmonella contamination. In the pre-harvest phase, this means that all available tools need to be used such as implementation of biosecurity measures, control of Salmonella infections in animals at the farm as well as in transport and trade, optimal housing and management including cleaning, disinfection procedures as well as efforts to achieve Salmonella-free feed production. This paper describes some nutritional strategies that could be used in farm control programmes in the major mono-gastric food production animals: poultry and pigs. Initially, it is important to prevent the introduction of Salmonella onto the farm through Salmonella-contaminated feed and this risk is reduced through heat treatment and the use of organic acids and their salts and formaldehyde. Microbiological sampling and monitoring for Salmonella in the feed mills is required to minimize the introduction of Salmonella via feed onto the farm. In addition, feed withdrawal may create a stressful situation in animals, resulting in an increase in Salmonella shedding. Physical feed characteristics such as coarse-ground meal to pigs can delay gastric emptying, thereby increasing the acidity of the gut and thus reducing the possible prevalence of Salmonella. Coarse-ground grains and access to litter have also been shown to decrease Salmonella shedding in poultry. The feed can also modify the gastro-intestinal tract microflora and influence the immune system, which can minimize Salmonella colonization and shedding. Feed additives, such as organic acids, short- and medium-chain fatty acids, probiotics, including competitive exclusion cultures, prebiotics and certain specific carbohydrates, such as mannan-based compounds, egg proteins, essential oils and bacteriophages, have the potential to reduce Salmonella levels when added to the feed. These nutritional strategies could be evaluated and used in farm control programmes. PMID:22436270

Berge, A C; Wierup, M

2012-04-01

191

Improved Isolation of Salmonellae from Naturally Contaminated Meat Products by Using Rappaport-Vassiliadis Enrichment Broth  

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A total of 454 specimens of meat products were examined for salmonellae by using five procedures of enrichment. The use of a selective motility medium, inoculated from enrichment in Muller-Kauffmann broth, resulted in an increase in the number of positive specimens. However, simple enrichment in Rappaport-Vassiliadis broth, after preenrichment, was more sensitive and specific for recovering salmonellae than the selective motility medium-Muller-Kauffmann broth method.

Vassiliadis, Peter; Kalapothaki, Victoria; Trichopoulos, Dimitrios; Mavrommatti, Christofili; Serie, Charles

1981-01-01

192

Identification of Two Phylogenetically Related Organisms from Feces by PCR for Detection of Salmonella spp.  

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Two previously reported PCR methods were evaluated to determine whether they are as sensitive and specific as conventional culture methods in detecting Salmonella spp. from feces. Bovine and equine feces were enriched overnight in brain heart infusion broth and assayed using PCR methods and primer sets described by other investigators. A total of 774 fecal specimens were tested using a primer set (invE-A primer set) that amplifies a region spanning the invasin E and A genes of Salmonella ente...

Gentry-weeks, Claudia; Hutcheson, H. Joel; Kim, Lisa Marie; Bolte, Denise; Traub-dargatz, Josie; Morley, Paul; Powers, Barbara; Jessen, Michael

2002-01-01

193

Salmonella: clinical importance and evolution of nomenclature.  

Science.gov (United States)

Salmonella is an important pathogen for both humans and animals. Although the organism has been intensively studied during the last century, much remains to be learned about this pathogen. The complicated nomenclature system of Salmonella has long been a subject of discussion. In 2005, "Salmonella enterica" finally gained official approval as the type species of the genus Salmonella. The genus Salmonella also contains the species "Salmonella bongori" in addition to a new species, "Salmonella subterranean", which was recognized in 2005. Unlike other bacterial genera, Salmonella organisms are differentiated by serotyping analysis. Presently, new serotypes (serovars) are still being discovered each year, adding to the complexity of this large bacterial population. Despite the conserved genetic background, molecular analysis has indicated successful evolution of the Salmonella genome in response to the environment, particularly to the selective pressure from antimicrobial agents. Mechanisms of fluoroquinolone resistance in Salmonella are similar to the complex system reported for other members of the family Enterobacteriaceae. On the other hand, resistance to extended-spectrum cephalosporins is more likely to be mediated by bla(CTX-M) or ampC genes that are carried on plasmids. Plasmid-borne genes have increased efficacy in the dissemination of resistance determinants, resulting in increased antimicrobial resistance. To provide clinicians with up-to-date information on this important pathogen, the evolving nomenclature and clinical importance of Salmonella are reviewed. PMID:17760271

Su, Lin-Hui; Chiu, Cheng-Hsun

2007-01-01

194

Abscesso esplênico causado por Salmonella / Splenic abscess due to Salmonella  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese OBJETIVO: Relatar as características demográficas, clínicas, diagnóstica e terapêutica de pacientes com abscesso esplênico (AE) causado por Salmonella. MÉTODO: Análise retrospectiva de dados de pacientes atendidos no Serviço de Cirurgia Geral e Aparelho Digestivo do Hospital Universitário Gaffrée-Gu [...] inle no período de janeiro de 2001 a dezembro de 2005, a estes se incluiu um caso tratado em outro hospital em época anterior. RESULTADOS: Dentre 4823 pacientes recentemente atendidos, dois apresentaram AE causado por Salmonella enteritidis, enquanto o caso mais antigo o agente responsável foi a Salmonella typhi. Todos eram homens, com idade média de 45 anos. Em nenhum deles foi identificada condição predisponente à formação do abscesso; os exames de imagem foram capazes de diagnosticar o AE. Todos foram tratados por esplenectomia e antibioticoterapia e evoluíram para cura. CONCLUSÕES: A Salmonella, apesar de infrequente, pode ser o agente causal do AE. Caracteristicamente os abscessos eram grandes e apresentavam material necrótico em seu interior. Nesta condição, a esplenectomia associada a antibioticoterapia mostrou-se eficaz no tratamento. Abstract in english BACKGROUND: Splenic abscess is a uncommon disease and remains a diagnostic challenge. Outcome is fatal when the condition is not promptly recognized and treated. The aim of this study was to analyze the demographic, clinical, diagnostic methods and management characteristic of patients with salmonel [...] al splenic abscess. METHODS: Retrospective study at General and Digestive Surgery Service - Hospital Universitário Gaffrée-Guinle of the Universidade Federal do Estado do Rio de Janeiro - UNIRIO from January 2001 to December 2005 resulting in a total number of 4823 patients has been performed. An additional case treated by one author was added. RESULTS: During the studied period two cases of splenic abscess due to Salmonella enteritidis was treated; a third older case was due to Salmonella typhi. All patients were man; the mean age was 45 years, all were immunocompetent and had large solitary lesion. All patients underwent splenectomy and antibiotic therapy; the outcome was favorable for all patients. CONCLUSION: Nowadays still is possible that splenic abscess is due to Salmonella species etiology; typically, all patients are immunocompetents adult males and large solitary lesions within necrotic material. In this condition, splenectomy and intravenous antibiotic administration appear to constitute the most convenient therapy.

Bruno von Glen, Herkenhoff; Ludmila Godoy dos Santos, Ferreira; Maristela, Cavedagne; Renato Manganelli, Salomão; Acyolli Moreira, Maia; Antonio Carlos, Iglesias.

195

A rabbit model of non-typhoidal Salmonella bacteremia.  

Science.gov (United States)

Bacteremia is an important cause of morbidity and mortality in humans. In this study, we focused on the development of an animal model of bacteremia induced by non-typhoidal Salmonella. New Zealand White rabbits were inoculated with a human isolate of non-typhoidal Salmonella strain CVD J73 via the intra-peritoneal route. Blood samples were collected at specific time points and at euthanasia from infected rabbits. Additionally, tissue samples from the heart, lungs, spleen, gastrointestinal tract, liver and kidneys were obtained at euthanasia. All experimentally infected rabbits displayed clinical signs of disease (fever, dehydration, weight loss and lethargy). Tissues collected at necropsy from the animals exhibited histopathological changes indicative of bacteremia. Non-typhoidal Salmonella bacteria were detected in the blood and tissue samples of infected rabbits by microbiological culture and real-time PCR assays. The development of this animal model of bacteremia could prove to be a useful tool for studying how non-typhoidal Salmonella infections disseminate and spread in humans. PMID:25033732

Panda, Aruna; Tatarov, Ivan; Masek, Billie Jo; Hardick, Justin; Crusan, Annabelle; Wakefield, Teresa; Carroll, Karen; Yang, Samuel; Hsieh, Yu-Hsiang; Lipsky, Michael M; McLeod, Charles G; Levine, Myron M; Rothman, Richard E; Gaydos, Charlotte A; DeTolla, Louis J

2014-09-01

196

Immunity to intestinal pathogens: lessons learned from Salmonella.  

Science.gov (United States)

Salmonella are a common source of food- or water-borne infection and cause a wide range of clinical disease in human and animal hosts. Salmonella are relatively easy to culture and manipulate in a laboratory setting, and the infection of laboratory animals induces robust innate and adaptive immune responses. Thus, immunologists have frequently turned to Salmonella infection models to expand understanding of host immunity to intestinal pathogens. In this review, I summarize current knowledge of innate and adaptive immunity to Salmonella and highlight features of this response that have emerged from recent studies. These include the heterogeneity of the antigen-specific T-cell response to intestinal infection, the prominence of microbial mechanisms to impede T- and B-cell responses, and the contribution of non-cognate pathways for elicitation of T-cell effector functions. Together, these different issues challenge an overly simplistic view of host-pathogen interaction during mucosal infection, but also allow deeper insight into the real-world dynamic of protective immunity to intestinal pathogens. PMID:24942689

McSorley, Stephen J

2014-07-01

197

Production and Kinetics of Salmonella enterica serovar enteritidis in Vibrofermentor  

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Full Text Available Food-borne human Salmonellosis caused by Salmonella enterica serovar enteritidis has given a rise to many of economic losses in terms of both poultry and food industries. Salmonella has a great importance among the major bacterial pathogens of poultry. In Turkey as well as all over the world prevention of Salmonella infection can be achieved by good monitoring and screening programs. More recently immunological test systems are used for diagnosis based on the detection of surface antigens such as lipopolysaccharide (LPS, flagellin etc. As to its production some different methods are used practically. In this work, the lab-scale production of Salmonella enterica serovar enteritidis was achieved both in shake-flask and vibrofermentor cultures and a comparative optimization of the process yield and production kinetics was carried out. The microorganisms were cultivated in brain heart infusion broth, at 37°C for 5 h. Specific growth rates and doubling times for both vibrofermentör and shake-flask were estimated as 0.509 and 0.709 h-1 and 75.0 and 58.6 min, respectively. LPS and flagella were extracted from lyophilized cultures.

A. Nalbantsoy

2007-01-01

198

Rational design of Salmonella-based vaccination strategies.  

Science.gov (United States)

A permanently growing body of information is becoming available about the quality of protective immune responses induced by mucosal immunization. Attenuated live bacterial vaccines can be administered orally and induce long-lasting protective immunity in humans without causing major side effects. An attenuated Salmonella enterica serovar Typhi strain is registered as live oral vaccine against typhoid fever and has been in use for more than two decades. Recombinant attenuated Salmonella strains are also an attractive means of delivering heterologous antigens to the immune system, thereby, stimulating strong mucosal and systemic immune responses and consequently provide an efficient platform technology to design novel vaccination strategies. This includes the choice of heterologous protective antigens and their expression under the control of appropriate promoters within the carrier strain. The availability of well-characterized attenuated mutants of Salmonella concomitantly supports fine tuning of immune response triggered against heterologous antigens. Exploring different mucosal sites as a potential route of immunization has to be taken into account as an additional important way to modulate immune responses according to clinical requirements. This article focuses on the rational design of strategies to modulate appropriate immunological effector functions on the basis of selection of (i) attenuating mutations of the Salmonella strains, (ii) specific expression systems for the heterologous antigens, and (iii) route of mucosal administration. PMID:16414270

Spreng, Simone; Dietrich, Guido; Weidinger, Gerald

2006-02-01

199

Prevalence, concentrations, and antibiotic sensitivities of Salmonella serovars in poultry from retail establishments in Seattle, Washington.  

Science.gov (United States)

Poultry have been identified as one of the major sources of salmonellosis, with estimates ranging from 10 to 22% of total cases. Despite several advances in the industry and new performance standards, the incidence of salmonellosis in the population has not declined over the last 15 years. Salmonella is pervasive in a wide variety of foods, and thus, estimating its burden resulting from specific food categories has been challenging and plagued with uncertainty due to critical data gaps. The objective of this study was to conduct a year-long market survey (1,322 samples) to help bridge the data gaps on the contamination rates and levels of Salmonella on raw poultry by product type (i.e., breast, thighs, drums, wings, and split breast) and production method (conventional versus organic). The isolates recovered were serotyped and tested for antibiotic sensitivities. A PCR method was utilized for initial screening of samples after an overnight enrichment in tryptic soy broth. Three-tube most-probable-number (MPN) assays and anti-Salmonella immunomagnetic separation methods were utilized to determine the levels of Salmonella and aid with the recovery of Salmonella species, respectively. Eleven percent of the samples were positive for Salmonella. Significant differences in percent positive rates by product type included up to a 4-fold difference in percent positive rates between establishments, ranging from 7 to 31%. Of the samples positive for Salmonella species, 94% had <30 MPN/100 g. Production methods identified as organic or as not using antibiotics had significantly higher rates of recovery of Salmonella. On the other hand, all of the Salmonella isolates that were resistant to two or more antibiotics originated from conventional processing establishments where antibiotics were utilized. In addition, a significant proportion of isolates from conventionally processed products were serotypes clinically relevant to humans. PMID:24853509

Mazengia, E; Samadpour, M; Hill, H W; Greeson, K; Tenney, K; Liao, G; Huang, X; Meschke, J S

2014-06-01

200

Contribution of the Type VI Secretion System Encoded in SPI-19 to Chicken Colonization by Salmonella enterica Serotypes Gallinarum and Enteritidis  

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Salmonella Gallinarum is a pathogen with a host range specific to poultry, while Salmonella Enteritidis is a broad host range pathogen that colonizes poultry sub-clinically but is a leading cause of gastrointestinal salmonellosis in humans and many other species. Despite recent advances in our understanding of the complex interplay between Salmonella and their hosts, the molecular basis of host range restriction and unique pathobiology of Gallinarum remain largely unknown. Type VI Secretion S...

Blondel, Carlos J.; Castro, Benjami?n; Chiang, Sebastia?n; Toro, Cecilia S.; Zaldi?var, Mercedes; Contreras, Ine?s; Andrews-polymenis, Helene L.; Santiviago, Carlos A.

2010-01-01

 
 
 
 
201

Eukaryotic signaling pathways targeted by Salmonella effector protein AvrA in intestinal infection in vivo  

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Full Text Available Abstract Background The Salmonella AvrA gene is present in 80% of Salmonella enterica serovar strains. AvrA protein mimics the activities of some eukaryotic proteins and uses these activities to the pathogen's advantage by debilitating the target cells, such as intestinal epithelial cells. Therefore, it is important to understand how AvrA works in targeting eukaryotic signaling pathways in intestinal infection in vivo. In this study, we hypothesized that AvrA interacts with multiple stress pathways in eukaryotic cells to manipulate the host defense system. A whole genome approach combined with bioinformatics assays was used to investigate the in vivo genetic responses of the mouse colon to Salmonella with or without AvrA protein expression in the early stage (8 hours and late stage (4 days. Specifically, we examined the gene expression profiles in mouse colon as it responded to pathogenic Salmonella stain SL1344 (with AvrA expression or SB1117 (without AvrA expression. Results We identified the eukaryotic targets of AvrA and the cell signaling pathways regulated by AvrA in vivo. We found that pathways, such as mTOR, NF-kappaB, platelet-derived growth factors, vascular endothelial growth factor, oxidative phosphorylation, and mitogen-activated protein kinase signaling are specifically regulated by AvrA in vivo and are associated with inflammation, anti-apoptosis, and proliferation. At the early stage of Salmonella infection, AvrA mainly targeted pathways related to nuclear receptor signaling and oxidative phosphorylation. At the late stage of Salmonella infection, AvrA is associated with interferon-gamma responses. Conclusion Both early and late phases of the host response exhibit remarkable specificity for the AvrA+ Salmonella. Our studies provide new insights into the eukaryotic molecular cascade that combats Salmonella-associated intestinal infection in vivo.

Wu Shaoping

2010-12-01

202

Cellulitis Due to Salmonella infantis.  

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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.

Satish R Patil

2013-01-01

203

Seroprevalence of Salmonella and Mycoplasma gallisepticum Infection in the Six Model Breeder Poultry Farms at Patuakhali District in Bangladesh  

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Full Text Available The present study was undertaken to know the seroprevalence of Salmonella and Mycoplasma gallisepticum (MG infection in six model breeder poultry farms (MBPFs located at kalapara Upazilla under Patuakhali district, Bangladesh. A total of 364 sera samples were collected from chickens belonging to six MBPFs. All sera samples were examined by rapid serum plate agglutination (SPA test using commercial Salmonella (SP and MG antigens to determine the presence of Salmonella and MG specific antibodies in different age and sex of birds belonging to MBPFs. In addition to that prevalence of Salmonella and Mycoplasma infection in MBPFs during rainy and winter seasons were also recorded. The results of serological tests were analyzed statistically. The overall prevalence of Salmonella and Mycoplasma infection in six MBPFs were recorded as 23.46% and 46.88% respectively. Prevalence of salmonella was recorded highest in rainy season (25.00% than the winter season (21.88. On the contrary, Mycoplasma infection was recorded highest in winter season (61.45% than the rainy season (51.74%. Both Salmonella and Mycoplasma infections were recorded highest in female birds (24.10% than the male birds (15.62%. The prevalence of MG infection decreased with the increase of age. MG infection recorded highest 71.42% at 18 weeks of age and lowest 50% at 22 weeks of age. On the other hand, the prevalence Salmonella infection was increased with the increase age. Salmonella infection was found highest 30.76% at 39 weeks of age and lowest 13.33% at 32 weeks of age. It was concluded from the present study that both Salmonella and MG infection were significantly present in all six MBPFs and SPA test could be used as a tool for quick detection of Salmonella and MG infection.

A.J. Sikder

2005-01-01

204

Same species, different diseases: how and why typhoidal and non-typhoidal Salmonella enterica serovars differ  

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Full Text Available Human infections by the bacterial pathogen Salmonella enterica represent major disease burdens worldwide. This highly ubiquitous species consists of more than 2600 different serovars that can be divided into typhoidal and non-typhoidal Salmonella (NTS serovars. Despite their genetic similarity, these two groups elicit very different diseases and distinct immune responses in humans. Comparative analyses of the genomes of multiple Salmonella serovars have begun to explain the basis of the variation in disease manifestations. Recent advances in modeling both enteric fever and intestinal gastroenteritis in mice will facilitate investigation into both the bacterial- and host-mediated mechanisms involved in salmonelloses. Understanding the genetic and molecular mechanisms responsible for differences in disease outcome will augment our understanding of Salmonella pathogenesis, host immunity, and the molecular basis of host specificity. This review outlines the differences in epidemiology, clinical manifestations, and the human immune response to typhoidal and NTS infections and summarizes the current thinking on why these differences might exist.

GuntramA.Grassl

2014-08-01

205

Analysis of Hexanitrostilbene (HNS) and Dipicryethane (DPE) for Mutagenicity by the Ames/Salmonella Assay  

Energy Technology Data Exchange (ETDEWEB)

The Ames/Salmonella assay, developed by Professor Bruce Ames at the University of California, Berkeley, is a rapid and sensitive assay for detecting mutagenicity of various chemical compounds (Maron and Ames, 1983). It is a widely accepted short-term assay for detecting chemicals that induce mutations in the histidine (his) gene of Salmonella typhimurium. This is a reverse mutation assay that detects the mutational reversion of his-dependent Salmonella to the his-independent counterpart. Thereby, mutagenic compounds will increase the frequency of occurrence of his-independent bacterial colonies. The assay utilizes the specific genetically constructed strains of bacteria either with or without mammalian metabolic activation enzymes (S9), Aroclor induced rat liver homogenate to assess the mutagenicity of different compounds. In this study, we will use the Ames/Salmonella assay to investigate the mutagenicity of Hexanitrostilbene (HNS) from both Bofors and Pantex, and Dipicryethane (DPE).

Wu, R; Felton, J

2007-10-12

206

76 FR 81513 - Guidance for Industry: Prevention of Salmonella  

Science.gov (United States)

...Guidance for Industry: Prevention of Salmonella Enteritidis in Shell Eggs During Production...industry entitled ``Prevention of Salmonella Enteritidis in Shell Eggs During Production...FDA's final rule ``Prevention of Salmonella Enteritidis in Shell Eggs During...

2011-12-28

207

75 FR 48973 - Draft Guidance for Industry: Prevention of Salmonella  

Science.gov (United States)

...Guidance for Industry: Prevention of Salmonella Enteritidis in Shell Eggs During Production...guidance entitled ``Prevention of Salmonella Enteritidis in Shell Eggs During Production...FDA's final rule ``Prevention of Salmonella Enteritidis in Shell Eggs During...

2010-08-12

208

9 CFR 113.122 - Salmonella Choleraesuis Bacterin.  

Science.gov (United States)

... 2010-01-01 2010-01-01 false Salmonella Choleraesuis Bacterin. 113.122 Section...Inactivated Bacterial Products § 113.122 Salmonella Choleraesuis Bacterin. Salmonella Choleraesuis Bacterin shall be prepared...

2010-01-01

209

40 CFR 79.68 - Salmonella typhimurium reverse mutation assay.  

Science.gov (United States)

...2010-07-01 2010-07-01 false Salmonella typhimurium reverse mutation assay...Requirements for Registration § 79.68 Salmonella typhimurium reverse mutation assay. (a) Purpose. The Salmonella typhimurium histidine (his)...

2010-07-01

210

9 CFR 113.120 - Salmonella Typhimurium Bacterin.  

Science.gov (United States)

... 2010-01-01 2010-01-01 false Salmonella Typhimurium Bacterin. 113.120 Section...Inactivated Bacterial Products § 113.120 Salmonella Typhimurium Bacterin. Salmonella Typhimurium Bacterin shall be prepared...

2010-01-01

211

9 CFR 113.123 - Salmonella Dublin Bacterin.  

Science.gov (United States)

... 2010-01-01 2010-01-01 false Salmonella Dublin Bacterin. 113.123 Section 113...Inactivated Bacterial Products § 113.123 Salmonella Dublin Bacterin. Salmonella Dublin Bacterin shall be prepared from a...

2010-01-01

212

40 CFR 79.68 - Salmonella typhimurium reverse mutation assay.  

Science.gov (United States)

...a) Purpose. The Salmonella typhimurium histidine...of the microorganism Salmonella typhimurium. (b...causes the addition or deletion of single or multiple...the DNA molecule. Salmonella typhimurium reverse...detects mutation in a gene of a...

2010-07-01

213

9 CFR 113.30 - Detection of Salmonella contamination.  

Science.gov (United States)

...2010-01-01 false Detection of Salmonella contamination. 113.30 Section...Procedures § 113.30 Detection of Salmonella contamination. The test for detection of Salmonella contamination provided in this...

2010-01-01

214

21 CFR 866.3550 - Salmonella spp. serological reagents.  

Science.gov (United States)

...2010-04-01 2010-04-01 false Salmonella spp. serological reagents. 866...Serological Reagents § 866.3550 Salmonella spp. serological reagents. (a) Identification. Salmonella spp. serological reagents...

2010-04-01

215

Salmonella enteritidis isolation from broiler chickens infected with low doses  

Directory of Open Access Journals (Sweden)

Full Text Available In this paper we present the possibilities for detection of Salmonella enteritidis (SE by bacteriological and serological technics. Also, we studied which tissue is the most suitable sample for Salmonella isolation. One day old and three weeks old broiler chickens were experimentally infected with low doses of SE. For the isolation of SE we used Rappaport Vassiliadis media. Specific antibodies in sera were detected using ELISA. According to our results, broiler chickens were susceptible to infection with low doses of SE. However, specific antibodies could not been found. SE was isolated from caeca, but not from livers of infected chickens when tested at the end of the experiment (at 6 weeks. Our results revealed that bacterial cultures are still the method of choice for detection of SE in broilers flocks.

Velhner Maja

2005-01-01

216

ABC medium, a new chromogenic agar for selective isolation of Salmonella spp.  

Science.gov (United States)

We describe a new chromogenic agar medium, ABC medium (alphabeta-chromogenic medium), which includes two substrates, 3, 4-cyclohexenoesculetin-beta-D-galactoside and 5-bromo-4-chloro-3-indolyl-alpha-D-galactopyranoside, to facilitate the selective isolation of Salmonella spp. This medium exploits the fact that Salmonella spp. may be distinguished from other members of the family Enterobacteriaceae by the presence of alpha-galactosidase activity in the absence of beta-galactosidase activity. A total of 1, 022 strains of Salmonella spp. and 300 other gram-negative strains were inoculated onto this medium. Of these, 1,019 (99.7%) strains of Salmonella spp. produced a characteristic green colony, whereas only 1 strain (0.33%) of non-Salmonella produced a green colony. A total of 283 stool samples were cultured onto desoxycholate citrate (DC) agar and ABC medium by direct inoculation and after selective enrichment in selenite broth. Overall, the sensitivity and specificity were superior for ABC medium (100 and 90.5%, respectively) than for DC agar (88 and 26.9%, respectively). We conclude that ABC medium offers a high degree of specificity for the detection of Salmonella spp. in stool samples. PMID:9986848

Perry, J D; Ford, M; Taylor, J; Jones, A L; Freeman, R; Gould, F K

1999-03-01

217

Salmonella transfer potential onto tomatoes during laboratory-simulated in-field debris removal.  

Science.gov (United States)

Florida Tomato Good Agricultural Practices (T-GAPs) mandate the removal of dirt and debris from tomatoes during harvest but do not provide any specific regulations or guidance; thus, the current practice of using cloths needs to be evaluated. This study examined Salmonella transfer from inoculated green tomatoes to uninoculated cloths and from inoculated cloths to uninoculated tomatoes, upon single and multiple touches. Tomatoes were spot inoculated with a rifampin-resistant Salmonella cocktail (10(7) CFU per tomato) and were touched with cloth (clean, dirty-dry, dirty-wet) at 0, 1, or 24 h postinoculation. Salmonella was enumerated on tryptic soy agar, followed by enrichments when necessary. The transfer direction was then reversed by touching freshly inoculated cloths with uninoculated tomatoes. Transfer coefficients (TCs) were then calculated. Salmonella TCs from inoculated tomato and cloth were highest when the inoculum was wet (0.44 ± 0.13 to 0.32 ± 0.12), regardless of the condition of the cloth. Although Salmonella TCs from inoculated tomato to uninoculated cloth decreased significantly when the inoculum was dried (0.17 ± 0.23 to 0.01 ± 0.00), low levels of Salmonella were detected on cloth even after 24 h of drying. Inoculated dirty cloth did not transfer more Salmonella compared with inoculated clean cloth, and Salmonella survival was not higher on dirty cloth. When inoculated clean cloth (wet) was touched with 25 tomatoes, significantly higher levels of Salmonella were transferred to the first, second, and fourth tomatoes (0.03 ± 0.10 to 0.09 ± 0.02). However, inoculated dirty-wet (below limit of detection) and dirty-dry (0.00 to 0.04 ± 0.01) cloths transferred similar levels of Salmonella to all 25 tomatoes. Results indicate a low risk of potential Salmonella contamination when the same cloth is used multiple times for debris removal, especially under high moisture levels. Results also show that the use of dirty cloths did not increase the risk of Salmonella cross-contamination. PMID:24988010

Sreedharan, Aswathy; Schneider, Keith R; Danyluk, Michelle D

2014-07-01

218

Antipathogenic activity of probiotics against Salmonella Typhimurium and Clostridium difficile in anaerobic batch culture systems: is it due to synergies in probiotic mixtures or the specificity of single strains?  

Science.gov (United States)

Probiotics are currently being investigated for prevention of infections caused by enteric pathogens. The aim of this in vitro study was to evaluate the influence of three single probiotics: Lactobacillus casei NCIMB 30185 (PXN 37), Lactobacillus acidophilus NCIMB 30184 (PXN 35), Bifidobacterium breve NCIMB 30180 (PXN 25) and a probiotic mixture containing the above strains plus twelve other strains belonging to the Lactobacillus, Bifidobacterium, Lactococcus, Streptococcus and Bacillus genera on the survival of Salmonella Typhimurium and Clostridium difficile using pH-controlled anaerobic batch cultures containing mixed faecal bacteria. Changes in relevant bacterial groups and effects of probiotic addition on survival of the two pathogens were assessed over 24 h. Quantitative analysis of bacterial populations revealed that there was a significant increase in lactobacilli and/or bifidobacteria numbers, depending on probiotic addition, compared with the control (no added probiotic). There was also a significant reduction in S. Typhimurium and C. difficile numbers in the presence of certain probiotics compared with controls. Of the probiotic treatments, two single strains namely L. casei NCIMB 30185 (PXN 37), and B. breve NCIMB 30180 (PXN 25) were the most potent in reducing the numbers of S. Typhimurium and C. difficile. In addition, the supplementation with probiotics into the systems influenced some fermentations parameters. Acetate was found in the largest concentrations in all vessels and lactate and formate were generally detected in higher amounts in vessels with probiotic addition compared to controls. PMID:24091275

Tejero-Sariñena, Sandra; Barlow, Janine; Costabile, Adele; Gibson, Glenn R; Rowland, Ian

2013-12-01

219

Salmonella outbreaks in restaurants in Minnesota, 1995 through 2003: evaluation of the role of infected foodworkers.  

Science.gov (United States)

The 23 restaurant-associated salmonellosis outbreaks that occurred in Minnesota from 1995 through 2003 were reviewed to characterize the role of infected foodworkers. The median duration of the outbreaks was 21 days (range, 1 to 517 days). The median number of culture-confirmed patron cases per outbreak was seven (range, 1 to 36 cases). The median incubation for patron cases ranged from 9 h to 5.9 days. A specific food vehicle was implicated in four outbreaks and suspected in five. Salmonella of the same serotype and pulsed-field gel electrophoresis subtype as that found in patrons was recovered from foodworkers in 19 outbreaks. Overall, 12% (129 of 1,033) of foodworkers tested positive for Salmonella. Sixty-four (53%) of 121 Salmonella-positive foodworkers reported not having had a recent gastrointestinal illness. Overall, the median duration of Salmonella shedding was 16 days. Among foodworkers who reported gastrointestinal illness, the median shedding duration was 30 days as compared with 3 days for asymptomatic foodworkers. Positive environmental samples were recovered in 4 (33%) of 12 outbreaks. No specific food vehicle was identified in any outbreaks associated with Salmonella-positive environmental samples. The median duration of outbreaks with positive environmental samples (187 days) was significantly longer than the median duration of outbreaks with negative environmental results (26 days, P = 0.03). A higher proportion of Salmonella-positive foodworkers (22 versus 8%) was identified in outbreaks with positive environmental samples. Salmonella outbreaks in restaurants are frequently prolonged yet produce a small number of confirmed patron cases. Prolonged outbreak durations suggest a persistent reservoir of contamination. Infected foodworkers likely serve as an important source for Salmonella transmission. Therefore, assessment of foodworker infection is essential for controlling restaurant outbreaks. PMID:16924912

Medus, Carlota; Smith, Kirk E; Bender, Jeffrey B; Besser, John M; Hedberg, Craig W

2006-08-01

220

Co-enriching microflora associated with culture based methods to detect Salmonella from tomato phyllosphere.  

Science.gov (United States)

The ability to detect a specific organism from a complex environment is vitally important to many fields of public health, including food safety. For example, tomatoes have been implicated numerous times as vehicles of foodborne outbreaks due to strains of Salmonella but few studies have ever recovered Salmonella from a tomato phyllosphere environment. Precision of culturing techniques that target agents associated with outbreaks depend on numerous factors. One important factor to better understand is which species co-enrich during enrichment procedures and how microbial dynamics may impede or enhance detection of target pathogens. We used a shotgun sequence approach to describe taxa associated with samples pre-enrichment and throughout the enrichment steps of the Bacteriological Analytical Manual's (BAM) protocol for detection of Salmonella from environmental tomato samples. Recent work has shown that during efforts to enrich Salmonella (Proteobacteria) from tomato field samples, Firmicute genera are also co-enriched and at least one co-enriching Firmicute genus (Paenibacillus sp.) can inhibit and even kills strains of Salmonella. Here we provide a baseline description of microflora that co-culture during detection efforts and the utility of a bioinformatic approach to detect specific taxa from metagenomic sequence data. We observed that uncultured samples clustered together with distinct taxonomic profiles relative to the three cultured treatments (Universal Pre-enrichment broth (UPB), Tetrathionate (TT), and Rappaport-Vassiliadis (RV)). There was little consistency among samples exposed to the same culturing medias, suggesting significant microbial differences in starting matrices or stochasticity associated with enrichment processes. Interestingly, Paenibacillus sp. (Salmonella inhibitor) was significantly enriched from uncultured to cultured (UPB) samples. Also of interest was the sequence based identification of a number of sequences as Salmonella despite indication by all media, that samples were culture negative for Salmonella. Our results substantiate the nascent utility of metagenomic methods to improve both biological and bioinformatic pathogen detection methods. PMID:24039862

Ottesen, Andrea R; Gonzalez, Antonio; Bell, Rebecca; Arce, Caroline; Rideout, Steven; Allard, Marc; Evans, Peter; Strain, Errol; Musser, Steven; Knight, Rob; Brown, Eric; Pettengill, James B

2013-01-01

 
 
 
 
221

Minimization of Salmonella contamination on raw poultry.  

Science.gov (United States)

Many reviews have discussed Salmonella in poultry and suggested best practices to minimize this organism on raw poultry meat. Despite years of research and conscientious control efforts by industry and regulatory agencies, human salmonellosis rates have declined only modestly and Salmonella is still found on raw poultry. Expert committees have repeatedly emphasized the importance of controlling risk, but information about Salmonella in poultry is often limited to prevalence, with inadequate information about testing methods or strains of Salmonella that are detected by these methods and no information about any impact on the degree of risk. This review examines some assumptions behind the discussion of Salmonella in poultry: the relationships between sampling and cultural methodology, prevalence and numbers of cells, and the implications of serotype and subtype issues. Minimizing Salmonella contamination of poultry is not likely to reduce human salmonellosis acquired from exposure to contaminated chicken until these issues are confronted more systematically. PMID:22129376

Cox, N A; Cason, J A; Richardson, L J

2011-01-01

222

Genome sequence of a salmonella phage used to control salmonella transmission in Swine.  

Science.gov (United States)

Salmonella shedding in swine often increases in response to transportation and lairage. We previously demonstrated that such increases can be limited by directly feeding microencapsulated Salmonella bacteriophages. Here we present the genome sequence of vB_SalM_SJ_3, a broader spectrum Viuna-like Salmonella phage used in those studies. PMID:25212610

Zhang, Jiayi; Hong, Yingying; Harman, Nicholas J; Das, Archana; Ebner, Paul D

2014-01-01

223

Oral priming with Salmonella Typhi vaccine strain CVD 909 followed by parenteral boost with the S. Typhi Vi capsular polysaccharide vaccine induces CD27+IgD-S. Typhi-specific IgA and IgG B memory cells in humans.  

Science.gov (United States)

Attenuated live oral typhoid vaccine candidate CVD 909 constitutively expresses Salmonella Typhi capsular polysaccharide antigen (Vi). A randomized, double-blind, heterologous prime-boost clinical study was conducted to determine whether immunity to licensed parenteral Vi vaccine could be enhanced by priming with CVD 909. Priming with CVD 909 elicited higher and persistent, albeit not significant, anti-Vi IgG and IgA following immunization with Vi, than placebo-primed recipients. Vi-specific IgA B memory (B(M)) cells were significantly increased in CVD 909-primed subjects. S. Typhi-specific LPS and flagella IgA B(M) cells were observed in subjects immunized with CVD 909 or with the licensed Vi-negative oral typhoid vaccine Ty21a. CVD 909-induced B(M) cells exhibited a classical B(M) phenotype (i.e., CD3(-)CD19(+)IgD(-)CD27(+)). This is the first demonstration of classical B(M) cells specific for bacterial polysaccharide or protein antigens following typhoid immunization. The persistent IgA B(M) responses demonstrate the capacity of oral typhoid vaccines to prime mucosally relevant immune memory. PMID:21146460

Wahid, Rezwanul; Pasetti, Marcela F; Maciel, Milton; Simon, Jakub K; Tacket, Carol O; Levine, Myron M; Sztein, Marcelo B

2011-02-01

224

Salmonella: Clinical Importance and Evolution of Nomenclature  

Directory of Open Access Journals (Sweden)

Full Text Available Salmonella is an important pathogen for both humans andanimals. Although the organism has been intensively studiedduring the last century, much remains to be learned about thispathogen. The complicated nomenclature system of Salmonellahas long been a subject of discussion. In 2005, “Salmonellaenterica” finally gained official approval as the type species ofthe genus Salmonella. The genus Salmonella also contains thespecies “Salmonella bongori” in addition to a new species,“Salmonella subterranean”, which was recognized in 2005.Unlike other bacterial genera, Salmonella organisms are differentiatedby serotyping analysis. Presently, new serotypes(serovars are still being discovered each year, adding to thecomplexity of this large bacterial population. Despite the conservedgenetic background, molecular analysis has indicatedsuccessful evolution of the Salmonella genome in response tothe environment, particularly to the selective pressure from antimicrobial agents.Mechanisms of fluoroquinolone resistance in Salmonella are similar to the complex systemreported for other members of the family Enterobacteriaceae. On the other hand, resistanceto extended-spectrum cephalosporins is more likely to be mediated by blaCTX-M or ampCgenes that are carried on plasmids. Plasmid-borne genes have increased efficacy in the disseminationof resistance determinants, resulting in increased antimicrobial resistance. Toprovide clinicians with up-to-date information on this important pathogen, the evolvingnomenclature and clinical importance of Salmonella are reviewed.

Cheng-Hsun Chiu

2007-06-01

225

Poly d,l-lactide-co-glycolide (PLGA) nanoparticle-encapsulated honeybee (Apis melifera) venom promotes clearance of Salmonella enterica serovar Typhimurium infection in experimentally challenged pigs through the up-regulation of T helper type 1 specific immune responses.  

Science.gov (United States)

Honeybee (Apis melifera) venom (HBV), which includes melittin and lipid-soluble ingredients (chrysin and pinocembrin), elicited increases in the CD4(+)/CD8(+) T lymphocyte ratio, relative mRNA expression levels of the T helper type 1 (Th 1) cytokines (interferon-? and IL-12) and reinforced viral clearance of an experimental porcine reproductive and respiratory syndrome (PRRS) virus infection in our previous study. On the basis of that previous study, we have now developed poly-d,l-lactide-co-glycolide (PLGA)-encapsulated HBV nanoparticles (P-HBV) for longer sustained release of HBV. We administered P-HBV to pigs via the rectal route, and then evaluated the potential immune-enhancing and bacterial clearance effects of P-HBV against Salmonella enterica serovar Typhimurium. The CD4(+)/CD8(+) lymphocyte ratio, proliferative capacity of peripheral blood lymphocytes and relative mRNA expression levels of IFN-? and IL-12 (produced mainly by Th1 lymphocytes) were significantly increased in the P-HBV group up to 2 weeks post-administration of P-HBV. After S. Typhimurium infection, the P-HBV group showed a marked reduction in microbial burden in feces and all tissue samples (including the ileum, cecum, colon, and mesenteric lymph node (MLN)), a significant increase in Th 1 cytokines (IFN-?, IL-2, and IL-12) and a marked decrease in a Th 2 cytokine (IL-4) in all tissue samples and peripheral blood lymphocytes. Thus, P-HBV may be a promising strategy for immune enhancement and prevention of S. Typhimurium or other bacterial infections. PMID:25193467

Lee, Jin-A; Jung, Bock-Gie; Kim, Tae-Hoon; Kim, Yun-Mi; Park, Min-Ho; Hyun, Pung-Mi; Jeon, Jong-Woon; Park, Jin-Kyu; Cho, Cheong-Weon; Suh, Guk-Hyun; Lee, Bong-Joo

2014-10-15

226

Shedding light on Salmonella carriers.  

Science.gov (United States)

Host-to-host transmission in most Salmonella serovars occurs primarily via the fecal-oral route. Salmonella enterica serovar Typhi is a human host-adapted pathogen and some S. Typhi patients become asymptomatic carriers. These individuals excrete large numbers of the bacteria in their feces and transmit the pathogen by contaminating water or food sources. The carrier state has also been described in livestock animals and is responsible for food-borne epidemics. Identification and treatment of carriers are crucial for the control of disease outbreaks. In this review, we describe recent advances in molecular profiling of human carriers and the use of animal models to identify potential host and bacterial genes involved in the establishment of the carrier state. PMID:22591832

Gopinath, Smita; Carden, Sarah; Monack, Denise

2012-07-01

227

Genetic Diversity of Salmonella Pathogenicity Islands SPI-5 and SPI-6 in Salmonella Newport.  

Science.gov (United States)

Abstract Salmonella enterica subspecies enterica serotype Newport is one of the common serotypes causing foodborne salmonellosis outbreaks in the United States. Salmonella Newport consists of three lineages exhibiting extensive genetic diversity. Due to the importance of Salmonella pathogenicity islands 5 and 6 (SPI-5 and SPI-6) in virulence of pathogenic Salmonella, the genetic diversity of these two SPIs may relate to different potentials of Salmonella Newport pathogenicity. Most Salmonella Newport strains from North America belong to Salmonella Newport lineages II and III. A total 28 Salmonella Newport strains of lineages II and III from diverse sources and geographic locations were analyzed, and 11 additional Salmonella genomes were used as outgroup in phylogenetic analyses. SPI-5 was identified in all Salmonella Newport strains and 146 single nucleotide polymorphisms (SNPs) were detected. Thirty-nine lineage-defining SNPs were identified, including 18 nonsynonymous SNPs. Two 40-kb genomic islands (SPI5-GI1 and SPI5-GI2) encoding bacteriophage genes were found between tRNA-ser and pipA. SPI5-GI1 was only present in Salmonella Newport multidrug-resistant strains of lineage II. SPI-6 was found in all strains but three Asian strains in Salmonella Newport lineage II, whereas the three Asian strains carried genomic island SPI6-GI1 at the same locus as SPI-6 in other Salmonella. SPI-6 exhibited 937 SNPs, and phylogenetic analysis demonstrated that clustering of Salmonella Newport isolates was a reflection of their geographic origins. The sequence diversity within SPI-5 and SPI-6 suggests possible recombination events and different virulence potentials of Salmonella Newport. The SNPs could be used as biomarkers during epidemiological investigations. PMID:25188010

Cao, Guojie; Allard, Marc; Strain, Errol; Stones, Robert; Zhao, Shaohua; Brown, Eric; Meng, Jianghong

2014-10-01

228

Calidad microbiológica y análisis de patógenos (Shigella y Salmonella) en lechuga / Microbiological quality and pathogen analysis (Shigella and Salmonella) of lettuce  

Scientific Electronic Library Online (English)

Full Text Available SciELO Venezuela | Language: Spanish Abstract in spanish Se analizó la calidad microbiológica en 37 muestras de lechuga variedad criolla (Lactuca sativa var. Capitata L.) de distintos intermediarios en las provincias de San José y Cartago, en Costa Rica. Las muestras se recolectaron mediante muestreo no probabilístico por selección intencional. Se cuantif [...] icó Escherichia coli (NMP/g) como indicador de contaminación fecal y se determinó la presencia de patógenos específicos (Shigella y Salmonella), por cultivo y por PCR-Múltiple. En el 65% de las muestras analizadas se detectó E. coli, aunque no se encontró Shigella ni Salmonella por PCR-Múltiple o cultivo. Una posible explicación es que los niveles de contaminación de Shigella y Salmonella están por debajo de los límites de detección de ambos métodos (menos de 10(4) UFC/g para Shigella y menos de 10² UFC/g para Salmonella). Estos resultados establecen una base importante para continuar con este tema de investigación y analizar otras fuentes de transmisión de Shigella y Salmonella, dado que ambos patógenos son frecuentes en la región. Abstract in english The microbiological quality of 37 lettuce samples of the creole variety (Lactuca sativa var. Capitata L.) obtained from different intermediaries at the provinces of San José and Cartago, in Costa Rica was analyzed. The samples were collected through a non-probabilistic sampling with intentional sele [...] ction. Escherichia coli (NMP/g) was quantified as indicator of fecal contamination and the presence of specific pathogens (Shigella and Salmonella) was determined by culture and Multiplex-PCR. In 65% of the samples analyzed we detected E. coli, even though we did not find Shigella or Salmonella by Multiplex-PCR or culture. A possible explanation is that the Shigella or Salmonella contamination levels may have been under the detection limits for both methods (less than 10(4) CFU/g for Shigella, and less than 10² CFU/g for Salmonella). These results establish an important basis for continuing with this research subject and analyzing other sources of transmission of Shigella and Salmonella contamination, since both pathogens are frequent in the region.

Kenia, Barrantes; Rosario, Achí.

229

Salmonella-like bioadhesive nanoparticles.  

Science.gov (United States)

The aim of this work was to evaluate the bioadhesive potential of a polymeric vector obtained by the association between Gantrez AN nanoparticles and flagella-enriched Salmonella enteritidis extract. Fluorescently labelled nanoparticles (SE-NP) were prepared, after incubation between the polymer and the extract, by a solvent displacement method and cross-linkage with 1,3-diaminopropane. SE-NP displayed a size close to 280 nm and the amount of associated bacterial extract was 18 mug/mg nanoparticle. Flagellin represents more than 80% of the total proteins associated with SE-NP, which was identified by SDS-PAGE and confirmed by Western blotting. Concerning the bioadhesive properties, SE-NP shows an important tropism for the ileum. In fact, about 50% of the given dose of SE-NP was found in this gut region for at least 3 h. Interestingly, the bioadhesive ability of SE-NP correlated well with the described colonisation profile for Salmonella enteritidis. This fact was corroborated by competitive tissue distribution studies. Thus, when SE-NP and Salmonella cells were administered together by the oral route, both the bacteria and the nanoparticles displayed a similar distribution within the intestinal mucosa. However, the ability of SE-NP to be taken up by Peyer's patches appeared to be negatively affected by the presence of the bacteria. Similarly, when SE-NP was administered 30 min before cells, SE-NP were found broadly distributed in Peyer's patches, whereas the bacteria were neither able to adhere to nor penetrate this lymphoid tissue. In summary, SE-NP demonstrated their Salmonella-like gut colonization, which can be a useful vector for oral targeting strategies. PMID:15970347

Salman, Hesham H; Gamazo, Carlos; Campanero, Miguel A; Irache, Juan M

2005-08-18

230

Meningoencefalitis letal por Salmonella B  

Digital Repository Infrastructure Vision for European Research (DRIVER)

La meningoencefalitis por bacilos gramnegativos ha ido incrementándose desde la década de los 70, con una mayor incidencia en niños pequeños, aunque existe una tendencia a aumentar en pacientes de la 3ra. edad. Dentro de este grupo de microorganismos, la causada por Salmonella sp, por su poca frecuencia, resulta una rareza. En este caso se presenta a una paciente de 80 años de edad con cuadro clínico de meningoencefalitis, que en el estudio del líquido cefalorraquídeo se aisló Salmon...

Herrera Valde?s, Nilda E.; Ma. Elena Fuerte Calvo; Ma. Elena Díaz García; Daysi Rodríguez Larrinaga; Martha Sandoval Acosta; Mario Santiago Puga Torres

2001-01-01

231

An allelotyping PCR for identifying Salmonella enterica serovars Enteritidis, Hadar, Heidelberg, and Typhimurium.  

Science.gov (United States)

Current commercial PCRs tests for identifying Salmonella target genes unique to this genus. However, there are two species, six subspecies, and over 2,500 different Salmonella serovars, and not all are equal in their significance to public health. For example, finding S. enterica subspecies IIIa Arizona on a table egg layer farm is insignificant compared to the isolation of S. enterica subspecies I serovar Enteritidis, the leading cause of salmonellosis linked to the consumption of table eggs. Serovars are identified based on antigenic differences in lipopolysaccharide (LPS)(O antigen) and flagellin (H1 and H2 antigens). These antigenic differences are the outward appearance of the diversity of genes and gene alleles associated with this phenotype. We have developed an allelotyping, multiplex PCR that keys on genetic differences between four major S. enterica subspecies I serovars found in poultry and associated with significant human disease in the US. The PCR primer pairs were targeted to key genes or sequences unique to a specific Salmonella serovar and designed to produce an amplicon with size specific for that gene or allele. Salmonella serovar is assigned to an isolate based on the combination of PCR test results for specific LPS and flagellin gene alleles. The multiplex PCRs described in this article are specific for the detection of S. enterica subspecies I serovars Enteritidis, Hadar, Heidelberg, and Typhimurium. Here we demonstrate how to use the multiplex PCRs to identify serovar for a Salmonella isolate. PMID:21808227

Maurer, John J; Lee, Margie D; Cheng, Ying; Pedroso, Adriana

2011-01-01

232

Evaluation of target sequences for the polymerase chain reaction-based detection of Salmonella in artificially contaminated beef.  

Science.gov (United States)

Salmonella is a major cause of foodborne diseases worldwide, which has fueled the demand for the development and evaluation of sensitive, specific, and rapid detection methodologies, such as polymerase chain reaction (PCR). In this study, six primer pairs for the detection of Salmonella were evaluated by PCR with isolates of Salmonella spp. (115) and other bacteria (104). The primers designed for the sifB gene provided the best performance regarding specificity and sensitivity (100%). These primers were selected and used to develop a PCR assay for Salmonella detection during the enrichment steps of the conventional detection method in spiked beef samples. The enrichment steps were: buffered peptone water (BPW), Rappaport-Vassiliadis soya broth (RVS) and at the Müller-Kauffmann tetrathionate novobiocin broth (MKTTn), after 18?h (BPW) and 24?h (RVS and MKTTn) of incubation. The initial concentrations of the Salmonella inocula were 10¹, 10², and 10³ colony-forming units/25 g. The protocol was able to detect Salmonella at all concentrations in the enrichment steps, but not in the nonenriched samples. These results indicated that the proposed protocol was suitable to detect Salmonella in beef during the intermediate stages of the conventional isolation protocol, substantially reducing the time required to obtain the final results. PMID:24102080

de Almeida, Michelle Vieira; Silva, Abelardo; Nero, Luís Augusto

2014-02-01

233

Development of Recombinant Flagellar Antigens for Serological Detection of Salmonella enterica Serotypes Enteritidis, Hadar, Heidelberg, and Typhimurium in Poultry  

Directory of Open Access Journals (Sweden)

Full Text Available Accurate and fast detection of harmful Salmonella is a major concern of food safety. Common Salmonella serotypes responsible for human associated foodborne outbreaks are S. Enteritidis, S. Hadar, S. Heidelberg, and S. Typhimurium are also commonly isolated from poultry. Serology is commonly used to monitor disease in poultry, therefore application of Salmonella serotype-specific test will have added value in Salmonella surveillance or monitoring vaccine efficacy. Recombinant flagellins were purified to be used as antigens in an ELISA. In this study, an ELISA was developed for the serological detection of S. Enteritidis. Once optimized, 500 ng of purified recombinant S. Enteritidis flagellin and a 1:64 dilution were determined to be optimal for testing sera. A negative baseline cutoff was calculated to be an optical density (OD of 0.35. All sera from birds with history of S. Enteritidis exposure tested positive and all sera from chickens with no exposure tested negative to this Salmonella serotype. Current ELISA for serological detection of Salmonella suffers from cross reactivity inherent in lipopolysaccharide (LPS or whole cell antigen based serological tests. This new ELISA eliminates common cross reactivity by focusing specifically on the flagellins of the Salmonella serotypes common in poultry and associated with foodborne outbreaks.

Charles L. Hofacre

2013-07-01

234

Development of an improved selective and differential medium for isolation of Salmonella spp.  

Science.gov (United States)

We describe an improved selective, differential, and cost-effective medium, XA medium, which contains d-arabinose, to facilitate the selective isolation of Salmonella spp. The sensitivity and the specificity of XA medium were compared to those of xylose lysine desoxycholate agar (XLD) using stock cultures and naturally contaminated food samples. XA medium and XLD were evaluated with a total of 82 Salmonella and 69 non-Salmonella stock cultures. Of 82 strains of Salmonella spp. tested, 76 produced a characteristic black colony on XA medium and XLD. The remaining 6 strains belonged to Salmonella enterica serovars Berta (n = 1), Paratyphi A (n = 1), Gallinarum (n = 2), and Pullorum (n = 2). The sensitivities of XA medium and XLD were identical (92.7%). Citrobacter freundii (n = 21) and Proteus mirabilis (n = 21) stock cultures produced black colonies on XLD, whereas only 4 strains of P. mirabilis appeared as black colonies on XA medium. In the second phase of the study, a total of 180 food samples were cultured onto XA medium and XLD after selective enrichment. The sensitivities of XA medium and XLD were equal (100%), and a total of 6 Salmonella strains were isolated from the 180 food samples. The specificity of XA medium (92.0%) was superior to that of XLD (73.0%), with a total of 14 and 47 false-positive results found on XA medium and XLD, respectively. On the basis of its good specificity, XA medium is useful for the isolation of Salmonella spp. from food samples. PMID:22814469

Park, Sang-Hyun; Ryu, Sangryeol; Kang, Dong-Hyun

2012-10-01

235

Spatio-temporal analysis of Salmonella surveillance data in Thailand  

DEFF Research Database (Denmark)

This study evaluates the usefulness of spatio-temporal statistical tools to detect outbreaks using routine surveillance data where limited epidemiological information is available. A dataset from 2002 to 2007 containing information regarding date, origin, source and serotype of 29 586 Salmonella isolates from Thailand was analysed. Data was grouped into human and non-human categories and the analysis was performed for the top five occurring serovars for each year of the study period. A total 91 human and 39 non-human significant spatio-temporal clusters were observed, accounting for 11% and 16% of the isolates, respectively. Serovar-specific associations between human and non-human clusters were also evaluated. Results show that these statistical tools can provide information for use in outbreak prevention and detection, in countries where only limited data is available. Moreover, it is suggested that monitoring non-human reservoirs can be relevant in predicting future Salmonella human cases.

Coutinho Calado Domingues, Ana Rita; Vieira, Antonio

2014-01-01

236

Sandwich enzyme immunoassays for detection of Salmonella typhi.  

Science.gov (United States)

Enzyme immunoassays were developed using monoclonal antibodies raised against somatic (O), flagellar (H) and capsular (Vi) antigens of Salmonella typhi. The assay based on anti-O monoclonal antibodies could specifically detect S. typhi and soluble lipopolysaccharide (LPS) isolated from S. typhi. Anti-H MoAbs detected motile S. typhi and soluble flagellar antigen. Monoclonal antibodies against capsular polysaccharide could detect Vi-containing S. typhi as well as soluble Vi antigen. The three assays reported here detected S. typhi with 100% sensitivity in blood culture broths obtained from bacteriologically confirmed typhoid patients and were negative with blood specimens containing Salmonella senftenberg, E. coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis or Streptococcus (alpha-hemolytic) derived from patients with pyrexia. The assays, however, did not demonstrate the presence of soluble antigens in sera and urine samples obtained from typhoid patients. PMID:1693382

Qadri, A; Ghosh, S; Prakash, K; Kumar, R; Moudgil, K D; Talwar, G P

1990-01-01

237

Salmonella infections: immune and non-immune protection with vaccines.  

Science.gov (United States)

Salmonella enterica in poultry remains a major political issue. S. enterica serovar Enteritidis, particularly, remains a world-wide problem. Control in poultry by immunity, whether acquired or innate, is a possible means of containing the problem. Widespread usage of antibiotics has led to the emergence of multiple antibiotic-resistant bacteria. This problem has indicated an increasing requirement for effective vaccines to control this important zoonotic infection. An attempt is made in the present review to explain the relatively poor success in immunizing food animals against these non-host-specific Salmonella serotypes that usually produce food-poisoning, compared with the success obtained with the small number of serotypes that more typically produce systemic "typhoid-like" diseases. New examinations of old problems such as the carrier state and vertical transmission, observed with S. Pullorum, is generating new information of relevance to immunity. Newer methods of attenuation are being developed. Live vaccines, if administered orally, demonstrate non-specific and rapid protection against infection that is of biological and practical interest. However, from the point of view of consumer safety, there is a school of thought that considers inactivated or sub-unit vaccines to be the safest. The benefits of developing effective killed or sub-unit vaccines over the use of live vaccines are enormous. Recently, there have been significant advances in the development of adjuvants (e.g. microspheres) that are capable of potent immuno-stimulation, targeting different arms of the immune system. The exploitation of such technology in conjunction with the ongoing developments in identifying key Salmonella virulence determinants should form the next generation of Salmonella sub-unit vaccines for the control of this important group of pathogens. There are additional areas of concern associated with the use of live vaccines, particularly if these are generated by genetic manipulation. PMID:17364505

Barrow, P A

2007-02-01

238

REGROWTH OF SALMONELLAE IN COMPOSTED SEWAGE SLUDGE  

Science.gov (United States)

Research was conducted to investigate the regrowth of salmonellae in composted sewage sludge. Though composting effectively stabilizes and disinfects sewage sludges, the decrease in salmonellae may be only temporary, since this pathogen can survive and grow without a human or ani...

239

Experimental Salmonella-associated conjunctivitis in cats.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Cats were infected experimentally with Salmonella typhimurium via the conjunctiva. Clinical signs consisted of lacrimation, conjunctivitis, blepharospasm, prominent nictitating membrane and scleral injection. These signs were accompanied by an absolute neutrophilia and conjunctival smears indicative of moderate to severe suppurative inflammation. Ocular signs disappeared by day 6 postinfection. Salmonella typhimurium was cultured intermittently from the inoculated conjunctivae and rectal swab...

Fox, J. G.; Beaucage, C. M.; Murphy, J. C.; Niemi, S. M.

1984-01-01

240

Salmonella bongori provides insights into the evolution of the Salmonellae.  

Science.gov (United States)

The genus Salmonella contains two species, S. bongori and S. enterica. Compared to the well-studied S. enterica there is a marked lack of information regarding the genetic makeup and diversity of S. bongori. S. bongori has been found predominantly associated with cold-blooded animals, but it can infect humans. To define the phylogeny of this species, and compare it to S. enterica, we have sequenced 28 isolates representing most of the known diversity of S. bongori. This cross-species analysis allowed us to confidently differentiate ancestral functions from those acquired following speciation, which include both metabolic and virulence-associated capacities. We show that, although S. bongori inherited a basic set of Salmonella common virulence functions, it has subsequently elaborated on this in a different direction to S. enterica. It is an established feature of S. enterica evolution that the acquisition of the type III secretion systems (T3SS-1 and T3SS-2) has been followed by the sequential acquisition of genes encoding secreted targets, termed effectors proteins. We show that this is also true of S. bongori, which has acquired an array of novel effector proteins (sboA-L). All but two of these effectors have no significant S. enterica homologues and instead are highly similar to those found in enteropathogenic Escherichia coli (EPEC). Remarkably, SboH is found to be a chimeric effector protein, encoded by a fusion of the T3SS-1 effector gene sopA and a gene highly similar to the EPEC effector nleH from enteropathogenic E. coli. We demonstrate that representatives of these new effectors are translocated and that SboH, similarly to NleH, blocks intrinsic apoptotic pathways while being targeted to the mitochondria by the SopA part of the fusion. This work suggests that S. bongori has inherited the ancestral Salmonella virulence gene set, but has adapted by incorporating virulence determinants that resemble those employed by EPEC. PMID:21876672

Fookes, Maria; Schroeder, Gunnar N; Langridge, Gemma C; Blondel, Carlos J; Mammina, Caterina; Connor, Thomas R; Seth-Smith, Helena; Vernikos, Georgios S; Robinson, Keith S; Sanders, Mandy; Petty, Nicola K; Kingsley, Robert A; Bäumler, Andreas J; Nuccio, Sean-Paul; Contreras, Inés; Santiviago, Carlos A; Maskell, Duncan; Barrow, Paul; Humphrey, Tom; Nastasi, Antonino; Roberts, Mark; Frankel, Gad; Parkhill, Julian; Dougan, Gordon; Thomson, Nicholas R

2011-08-01

 
 
 
 
241

Hemagglutinating properties of Salmonella enterica serovar Enteritidis isolated from different sources Propriedades hemaglutinantes de Salmonella enterica sorotipo Enteritidis isoladas de diferentes fontes  

Directory of Open Access Journals (Sweden)

Full Text Available Twenty-five strains of Salmonella enterica serovar Enteritidis isolated from different sources were examined for hemagglutinating activity. Bacteria cultured in different media induced hemagglutination of human erythrocytes, but no reaction was observed with erythrocytes from other animal species. The hemagglutinating expression activity was better for cultures on CFA agar at 37ºC than other conditions examined. The hemagglutination was inhibited by D-mannose, D-mannitol, melibiose, D-raffinose, L-rhamnose and sucrose. The absence of cell-surface appendages in electron microscope examinations suggested a nonfimbrial hemagglutinin. The data suggest that Salmonella Enteritidis produces nonfimbrial mannose-sensitive hemagglutinin, specific for human erythrocytes, which could be extracted in soluble form.Foram estudadas 25 amostras de Salmonella enterica sorotipo Enteritidis isoladas de diferentes fontes, em testes de hemaglutinação. Amostras bacterianas cultivadas em diferentes meios de cultura causavam hemaglutinação na presença de hemácias humanas, entretanto, não foi observada reação com hemácias de outras espécies. A expressão da atividade hemaglutinante foi melhor em ágar CFA a 37ºC. A hemaglutinação foi inibida por D-manose, D-manitol, melibiose, D-rafinose, L-ramnose e sacarose. A análise ultraestrutural não revelou a presença de estruturas filamentosas na superfície bacteriana, sugerindo que a hemaglutinina de Salmonella Enteritidis seja de natureza não fimbrial. Os dados sugerem que Salmonella Enteritidis produz uma hemaglutinina não fimbrial manose-sensível, específica para hemácias humanas, que pode ser extraída na forma solúvel.

Jane M.G. Mikcha

2004-06-01

242

Pathogenesis of Salmonella-induced enteritis  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Infections with Salmonella serotypes are a major cause of food-borne diseases worldwide. Animal models other than the mouse have been employed for the study of nontyphoidal Salmonella infections because the murine model is not suitable for the study of Salmonella-induced diarrhea. The microbe has de [...] veloped mechanisms to exploit the host cell machinery to its own purpose. Bacterial proteins delivered directly into the host cell cytosol cause cytoskeletal changes and interfere with host cell signaling pathways, which ultimately enhance disease manifestation. Recently, marked advances have been made in our understanding of the molecular interactions between Salmonella serotypes and their hosts. Here, we discuss the molecular basis of the pathogenesis of Salmonella-induced enteritis.

R.L., Santos; R.M., Tsolis; A.J., Bäumler; L.G., Adams.

2003-01-01

243

Pathogenesis of Salmonella-induced enteritis  

Directory of Open Access Journals (Sweden)

Full Text Available Infections with Salmonella serotypes are a major cause of food-borne diseases worldwide. Animal models other than the mouse have been employed for the study of nontyphoidal Salmonella infections because the murine model is not suitable for the study of Salmonella-induced diarrhea. The microbe has developed mechanisms to exploit the host cell machinery to its own purpose. Bacterial proteins delivered directly into the host cell cytosol cause cytoskeletal changes and interfere with host cell signaling pathways, which ultimately enhance disease manifestation. Recently, marked advances have been made in our understanding of the molecular interactions between Salmonella serotypes and their hosts. Here, we discuss the molecular basis of the pathogenesis of Salmonella-induced enteritis.

Santos R.L.

2003-01-01

244

Salmonella bongori provides insights into the evolution of the Salmonellae  

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The genus Salmonella contains two species, S. bongori and S. enterica. Compared to the well-studied S. enterica there is a marked lack of information regarding the genetic makeup and diversity of S. bongori. S. bongori has been found predominantly associated with cold-blooded animals, but it can infect humans. To define the phylogeny of this species, and compare it to S. enterica, we have sequenced 28 isolates representing most of the known diversity of S. bongori. This cross-species analysis...

Fookes, M.; Schroeder, G. N.; Langridge, G. C.; Blondel, C. J.; Mammina, C.; Connor, T. R.; Seth-smith, H.; Vernikos, G. S.; Robinson, K. S.; Sanders, M.; Petty, N. K.; Kingsley, R. A.; Ba?umler, A. J.; Nuccio, S. -p; Contreras, I.

2011-01-01

245

Non-Typhoidal Salmonella Aortitis in a transplant patient  

International Nuclear Information System (INIS)

Non-typhoidal salmonella bacteremia may result in extra gastrointestinallocalization of infection. Aortitis due to non-typhoidal salmonella wasreported to be the cause of 38-42% of all infected abdominal aortitis.Underlying atherosclerosis is a frequent site for salmonella aortitis. Wedescribe here a case of possible salmonella aortitis in a renal transplantpatient. (author)

246

16-23S rRNA Spacer Region Polymorphism in Gangetic River Water Isolates of Salmonella  

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Full Text Available Salmonella is one of the major pathogenic bacteria present in contaminated water. 16-23S rRNA spacer region has been reported to be polymorphic at serovar level in Salmonella. Salmonella isolates obtained from Ganges river water were studied for 16-23S rRNA spacer region polymorphism. Thirty three isolates belonging to eight serovars (S. Typhimurium, S. Abuja, S. Pantypridd, S. Lagos, S. Chinkual, S. Zwickau, S. Goldenberg and S. Oritamerin were studied for the polymorphism. Out of 33 isolates, 15 different profiles were observed no serovar specific profile. Our findings indicate that 16-23S rRNA spacer region is not specific at serovar level, but can be used for differentiation of different Salmonella isolates.

Rubi Singh

2010-08-01

247

Protective effect of probiotics on Salmonella infectivity assessed with combined in vitro gut fermentation-cellular models  

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Full Text Available Abstract Background Accurate assessment of probiotics with targeted anti-Salmonella activity requires suitable models accounting for both, microbe-microbe and host-microbe interactions in gut environments. Here we report the combination of two original in vitro intestinal models closely mimicking the complex in vivo conditions of the large intestine. Effluents from continuous in vitro three-stage fermentation colonic models of Salmonella Typhimurium infection inoculated with immobilized child microbiota and Salmonella were directly applied to confluent mucus-secreting HT29-MTX cell layers. The effects of Salmonella, addition of two bacteriocinogenic strains, Bifidobacterium thermophilum RBL67 (thermophilicin B67 and Escherichia coli L1000 (microcin B17, and inulin were tested on Salmonella growth and interactions with epithelial cell layers. Salmonella adhesion and invasion were investigated and epithelial integrity assessed by transepithelial electrical resistance (TER measurements and confocal microscopy observation. Data from complex effluents were compared with pure Salmonella cultures. Results Salmonella in effluents of all reactors of the colonic fermentation model stabilized at mean values of 5.3 ± 0.8 log10 cfu/ml effluent. Invasion of cell-associated Salmonella was up to 50-fold lower in complex reactor samples compared to pure Salmonella cultures. It further depended on environmental factors, with 0.2 ± 0.1% being measured with proximal, 0.6 ± 0.2% with transverse and 1.3 ± 0.7% with distal reactor effluents, accompanied by a similar high decrease of TER across cell monolayers (minus 45% and disruption of tight junctions. Subsequent addition of E. coli L1000 stimulated Salmonella growth (6.4 ± 0.6 log10 cfu/ml effluent of all 3 reactors and further decreased TER, but led to 10-fold decreased invasion efficiency when tested with distal reactor samples. In contrast, presence of B. thermophilum RBL67 revealed a protective effect on epithelial integrity compared to previous E. coli L1000 periods, as reflected by a significant mean increase of TER by 58% in all reactors. Inulin addition enhanced Salmonella growth and invasion when tested with distal and proximal reactor samples, respectively, but induced a limited decrease of TER (minus 18% in all reactors. Conclusions Our results highlight the benefits of combining suitable cellular and colonic fermentation models to assess strain-specific first-level host protection properties of probiotics during Salmonella infection, providing an efficient system biology tool for preclinical development of new antimicrobials.

Zihler Annina

2011-12-01

248

A colanic acid operon deletion mutation enhances induction of early antibody responses by live attenuated Salmonella vaccine strains.  

Science.gov (United States)

Colanic acid (CA) is a common exopolysaccharide produced by many genera in the Enterobacteriaceae. It is critical for biofilm formation on HEp-2 cells and on chicken intestinal tissue by Salmonella. In this study, we generated different CA synthesis gene mutants and evaluated the immune responses induced by these mutants. One of these mutations, ?(wza-wcaM)8, which deleted the whole operon for CA synthesis, was introduced into two Salmonella vaccine strains attenuated by auxotrophic traits or by the regulated delayed attenuation strategy (RDAS). The mice immunized with the auxotrophic Salmonella vaccine strain with the deletion mutation ?(wza-wcaM)8 developed higher vaginal IgA titers against the heterologous protective antigen and higher levels of antigen-specific IgA secretion cells in lungs. In Salmonella vaccine strains with RDAS, the strain with the ?(wza-wcaM)8 mutation resulted in higher levels of protective antigen production during in vitro growth. Mice immunized with this strain developed higher serum IgG and mucosal IgA antibody responses at 2 weeks. This strain also resulted in better gamma interferon (IFN-?) responses than the strain without this deletion at doses of 10(8) and 10(9) CFU. Thus, the mutation ?(wza-wcaM)8 will be included in various recombinant attenuated Salmonella vaccine (RASV) strains with RDAS derived from Salmonella enterica serovar Paratyphi A and Salmonella enterica serovar Typhi to induce protective immunity against bacterial pathogens. PMID:23774599

Wang, Shifeng; Shi, Huoying; Li, Yuhua; Shi, Zhaoxing; Zhang, Xin; Baek, Chang-Ho; Mothershead, Tabor; Curtiss, Roy

2013-09-01

249

The Deubiquitinase Activity of the Salmonella Pathogenicity Island 2 Effector, SseL, Prevents Accumulation of Cellular Lipid Droplets ? †  

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To cause disease, Salmonella enterica serovar Typhimurium requires two type III secretion systems that are encoded by Salmonella pathogenicity islands 1 and 2 (SPI-1 and -2). These secretion systems serve to deliver specialized proteins (effectors) into the host cell cytosol. While the importance of these effectors to promote colonization and replication within the host has been established, the specific roles of individual secreted effectors in the disease process are not well understood. In...

Arena, Ellen T.; Auweter, Sigrid D.; Antunes, L. Caetano M.; Vogl, A. Wayne; Han, Jun; Guttman, Julian A.; Croxen, Matthew A.; Menendez, Alfredo; Covey, Scott D.; Borchers, Christoph H.; Finlay, B. Brett

2011-01-01

250

Salmonella radicidation of poultry carcasses  

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Validity of methodsExperiments were carried out In which it was assessed which Salmonella isolation method is the most productive one In the examination of broiler carcasses. Refrigerated, refrigerated and radiated (2.50 kGy), frozen and frozen and radiated (2.50 kGy) samples of broilers were examined. After evaluation of all results It was concluded that the following method was the most productive one:1. pre-enrichment in buffered peptone water at 37 °C for 20 hours2. enrichm...

Mulder, R. W. A. W.

1982-01-01

251

Significance of Salmonella typhi bacteriuria.  

Science.gov (United States)

Bacteriuria due to Salmonella typhi usually occurs following recent typhoid fever or in chronic carrier states. Data from 18 patients with S. typhi bacteriuria, seen during 5 years, were analyzed. Fourteen patients had localized urinary tract infection due to S. typhi. Four others had bacteriuria, probably associated with typhoid fever. Localized abnormalities of the urinary tract and kidneys and also systemic diseases were found to predispose patients to S. typhi bacteriuria. Local abnormalities encountered included urolithiasis (n = 3), prostatic hypertrophy (n = 1), and tuberculosis (n = 1). One renal transplant recipient and another with lupus nephritis had S. typhi bacteriuria. One had associated strongyloidosis, and another was pregnant. PMID:7545180

Mathai, E; John, T J; Rani, M; Mathai, D; Chacko, N; Nath, V; Cherian, A M

1995-01-01

252

Significance of Salmonella typhi bacteriuria.  

Science.gov (United States)

Bacteriuria due to Salmonella typhi usually occurs following recent typhoid fever or in chronic carrier states. Data from 18 patients with S. typhi bacteriuria, seen during 5 years, were analyzed. Fourteen patients had localized urinary tract infection due to S. typhi. Four others had bacteriuria, probably associated with typhoid fever. Localized abnormalities of the urinary tract and kidneys and also systemic diseases were found to predispose patients to S. typhi bacteriuria. Local abnormalities encountered included urolithiasis (n = 3), prostatic hypertrophy (n = 1), and tuberculosis (n = 1). One renal transplant recipient and another with lupus nephritis had S. typhi bacteriuria. One had associated strongyloidosis, and another was pregnant. PMID:7545180

Mathai, E; John, T J; Rani, M; Mathai, D; Chacko, N; Nath, V; Cherian, A M

1995-07-01

253

Optical immunosensors for detection of Listeria monocytogenes and Salmonella enteritidis from food  

Science.gov (United States)

Listeria monocytogenes and Salmonella are two major foodborne pathogens of significant concern. Two optical evanescent wave immunosensors were evaluated for detection: Antibody-coupled fiber-optic biosensor and a surface plasmon resonant (SPR) immunosensor. In the fiber-optic sensor, polyclonal antibodies for the test organisms were immobilized on polystyrene fiber wave -guides using streptavidin - biotin chemistry. Cyanine 5 -labeled monoclonal antibodies C11E9 (for L. monocytogenes) and SF-11 (for Salmonella Enteritidis) were used to generate a specific fluorescent signal. Signal acquisition was performed by launching a laser-light (635 nm) from an Analyte-2000. This immunosensor was able to detect 103 - 109 cfu/ml of L. monocytogenes or 106-109 cfu/ml of Salmonella Enteritidis and the assays were conducted at near real-time with results obtained within one hour of sampling. The assays were specific and showed signal even in the presence of other microorganisms such as E. coli, Enterococcus faecalis or Salmonella Typhimurium. In the SPR system, IAsys instrument (resonant mirror sensor) was used. Monoclonal antibody-C11E9 was directly immobilized onto a carboxylate cuvette. Whole Listeria cells at various concentrations did not yield any signal while surface protein extracts did. Crude protein extracts from L. monocytogenes and L. innocua had average binding responses of around 150 arc sec (0.25 ng/mm2), which was significantly different from L. grayi, L. ivanovii, or L. welshimeri with average responses of monocytogenes and Salmonella Enteritidis.

Bhunia, Arun K.; Geng, Tao; Lathrop, Amanda; Valadez, Angela; Morgan, Mark T.

2004-03-01

254

A comparative study of culture methods and polymerase chain reaction assay for Salmonella detection in poultry feed.  

Science.gov (United States)

The present work compared 2 culture methods and PCR assay for the detection of motile and non-motile Salmonella strains using artificially contaminated poultry feed. The specificity was 1 in all methods. The accuracy and sensitivity were between 0.5 and 1 for motile Salmonella strains, whereas these parameters were between 0 and 0.6 for non-motile Salmonella strains. The positive predictive value was 1 for tetrathionate (TT), PCR, and modified semisolid Rappaport-Vassiliadis (MSRV) methods in most of the strains studied. The negative predictive value of each method was very low for non-motile Salmonella strains. The detection level of motile strains was 8 to 20 cfu/25 g for all methods, whereas it was ?10(4) cfu/25 g in culture methods for non-motile Salmonella strains. In general, the PCR method detected lower non-motile Salmonella contamination levels in feed than did culture methods. Extending incubation time of the enrichment medium to 6 d in the TT method did not improve the isolation rates. All selective plating media did not show any statistical differences in the parameters of performance studied. Kappa coefficients showed that there was good agreement between TT and MSRV methods, and MSRV and PCR methods for motile Salmonella strains in poultry feed samples. The agreement was fair between TT and PCR methods for these strains. For non-motile Salmonella strains, there was poor (TT and MSRV methods), slight (PCR and TT methods), and fair (MSRV and PCR methods) agreement. The TT, MSRV, and PCR methods are similar in terms of accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for different motile Salmonella strains in poultry feed. For non-motile Salmonella strains, the use of the PCR method improves the same parameters, described before, in this matrix. The difference in detection levels obtained with the methods used for motile and nonmotile Salmonella strains and the difficulty to detect these last strains represent a potential problem, when a poultry feed sample is considered negative for the presence of Salmonella. PMID:22010248

Soria, M C; Soria, M A; Bueno, D J; Colazo, J L

2011-11-01

255

Occurrence of Salmonella sp in laying hens  

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Full Text Available This study was carried out to investigate the presence of Salmonella sp in flocks of white laying hens. In different farms, the transport boxes of twelve flocks were inspected at arrival for the presence of Salmonella. Four positive (A, B, L and M and one negative (I flocks were monitored at each four weeks using bacteriological examination of cecal fresh feces up to 52 weeks. Birds were also evaluated at 52 weeks, when 500 eggs were taken randomly, and at 76 weeks, after forced molt. Salmonella enterica serovar Enteritidis and S. enterica rough strain were isolated from the transport boxes of the four positive flocks (flocks A, B, L and M. Salmonella sp was not isolated from the transport boxes or from the feces after 76 weeks-old in flock I. Salmonella sp was isolated in the 1st, 11th, 34th, 42nd and 76th weeks from flock A; in the 1st, 4th, 11th and 76th weeks from flock B; in the first week and in the 17th to 52nd weeks from flock L; and in the 1st and 76th weeks from flock M. S. Enteritidis, S. enterica rough strain and Salmonella enterica serovar Infantis were isolated from the four positive flocks. Besides, Salmonella enterica serovar Javiana was isolated from flocks B and L, and Salmonella enterica serovar Mbandaka was isolated from flock L. Eggs produced by flock A and by flock L were contaminated with S. Enteritidis and S. enterica rough strain. According to these results, Salmonella-infected flocks may produce contaminated eggs.

Gama NMSQ

2003-01-01

256

Evaluation of associations between feed withdrawal and other management factors with Salmonella contamination of broiler chickens at slaughter in Alberta.  

Science.gov (United States)

Salmonellosis is one of the most common bacterial foodborne diseases of public health concern in industrialized countries. Poultry products are considered an important source of Salmonella-related foodborne disease in humans. This study was undertaken to evaluate the relationship between various management factors including feed withdrawal and transportation time with Salmonella contamination in crops, ceca, and carcasses of broiler chickens at slaughter in Alberta. Using a two-stage sampling procedure, 30 matched crop and cecal samples before evisceration and an additional 30 neck skin samples after final wash of broiler chickens were collected at slaughter. A questionnaire was administered at the time of sampling to collect information on flock management risk factors. Cecal contents were individually screened with Salmonella-specific real-time PCR to detect positive flocks, and all cecal, crop, and neck skin samples from positive flocks were processed further for Salmonella isolation and characterization. The flock prevalence of Salmonella was 57.1% and within-flock prevalence of Salmonella for positive flocks was 17.2, 8.1, and 53.9% for ceca, crops, and neck skins, respectively. Salmonella Hadar was the most common serovar identified from crops, ceca, and neck skins of broiler chickens tested. Longer transport (P = 0.04 for neck skins) and waiting time in-plant (P = 0.04 for crops, P = 0.03 for ceca) were identified as important risk factors for Salmonella contamination of broiler chickens at slaughter. Salmonella contamination of broiler chickens could potentially be minimized by reducing waiting time in-plant for flocks with longer transport time. PMID:19833047

Mainali, C; Gensler, G; McFall, M; King, R; Irwin, R; Senthilselvan, A

2009-10-01

257

Short communication: Determination of Salmonella clustered regularly interspaced short palindromic repeats (CRISPR) diversity on dairy farms in Wisconsin and Minnesota.  

Science.gov (United States)

Salmonella enterica ssp. enterica is a foodborne pathogen able to cause disease in both humans and animals. Diverse serovars of this pathogen exist, some of which are host specific, causing a range of clinical symptoms from asymptomatic infection through morbidity and mortality. According to a 2007 survey by the USDA National Animal Health Monitoring System, fecal shedding of Salmonella from healthy cows occurs on 39.7% of dairy farms in the United States. Certain serovars are frequently isolated from dairy farms and the majority of isolates from the National Animal Health Monitoring System study were represented by 5 serovars; however, genotypic diversity was not examined. The objective of this study was to determine the diversity of clustered regularly interspaced short palindromic repeats (CRISPR) loci in Salmonella collected from 8 dairy farms with a previous history of salmonellosis. None of the cows or calves sampled on 2 of the 8 dairy farms were shedding Salmonella, although Salmonella was detected in a cow bedding sample on 1 of these farms. Salmonella populations were discrete on each farm, according to CRISPR typing, with the exception of an Anatum var. 15+ type on farms 5 and 6 and the Montevideo type on farms 1 and 2. One to 4 distinct CRISPR genotypes were identified per farm. The CRISPR typing differed within serovars, as Montevideo, Anatum var. 15+, and Muenster serovars had no overlap of spacer content, even on the same farm, reflecting between- and within-serovar genetic diversity. The dynamic nature of Salmonella populations was shown in a farm that was sampled longitudinally over 13.5mo. Changes in serovar from 3,19:-:z27 to Montevideo was observed between the first sampling time and 8mo later, with concomitant change in CRISPR alleles. The results indicate that Salmonella strains present in smaller dairy herds (farm and new Salmonella strains may emerge over time. PMID:25108866

Wehnes, C A; Rehberger, T G; Barrangou, R; Smith, A H

2014-10-01

258

Comparison of CHROMagar Salmonella Medium and Hektoen Enteric Agar for Isolation of Salmonellae from Stool Samples  

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CHROMagar Salmonella (CAS), a new chromogenic medium, was retrospectively compared to Hektoen enteric agar (HEA) with 501 Salmonella stock isolates and was then prospectively compared to HEA for the detection and presumptive identification of Salmonella spp. with 508 stool samples before and after enrichment. All stock cultures (100%), including cultures of H2S-negative isolates, yielded typical mauve colonies on CAS, while 497 (99%) isolates produced typical lactose-negative, black-centered ...

Gaillot, Olivier; Di Camillo, Patrick; Berche, Patrick; Courcol, Rene?; Savage, Colette

1999-01-01

259

Comparison of Salmonella Chromogenic Medium with DCLS Agar for Isolation of Salmonella Species from Stool Specimens  

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Salmonella chromogenic medium (SCM; Oxoid, Basingstoke, United Kingdom), a new selective chromogenic medium, was compared to DCLS agar (Oxoid) for the detection and presumptive identification of Salmonella species from stool samples. This medium contains two chromogenic substrates, Magenta-cap (5-bromo-6-chloro-3-indolylcaprylate), which is hydrolyzed by Salmonella species to give magenta colonies, and X-Gal (5-bromo-4-chloro-3-indolyl-?-d-galactopyranoside), which is incorporated to visuali...

Cassar, Robert; Cuschieri, Paul

2003-01-01

260

Functional and phenotypic profiling of innate immunity during Salmonella infection  

DEFF Research Database (Denmark)

Salmonellae are food borne pathogens, typically acquired by the oral ingestion of contaminated food or water, causing disease in both healthy and immunocompromised individuals. To gain insight into early immune regulation events caused by Salmonella as well as inflammatory signatures induced by Salmonella and other bacteria in human monocyte-derived dendritic cells (DC), we examined these properties using in vivo and in vitro experimental settings. The outcome of infection with Salmonella depends on the host as well as the infecting serovar. Understanding the relative risks associated with and within different serovars is of major importance for public health. Using an established mouse model, we compared the pathogenicity of two S. Typhimurium strains (SL1344 and DT120) and found that the passage through and the ability to proliferate within the host gastrointestinal system determined the pathogenicity of these strains. Salmonella is a mucosal pathogen, gaining access to host systemic circulation by crossing the gut epithelial barrier and residing intracellularly in DC and M?. Until recently focus has been centred on the involvement of M? and the conventional antigen-presenting DC (mDC) in bacterial infections, whereas the other major dendritic cell subset, plasmacytoid DC (pDC), plays an important part in antiviral responses, and is less well characterised in regard to antibacterial immunity. Using multi-parametric flow cytometry, we were able to show for the first time that pDC accumulated in Peyer’s patches 24 hours after murine oral Salmonella challenge and while M? and mDC exhibited dose-related cellular atrophy, pDC were less susceptible to bacteria-induced cell death, suggesting a role for pDC in early stage Salmonella containment. Furthermore, we identified a number of both DC and M? subsets, two of which following infection, accumulated in Peyer’s patches and lamina propria, respectively. Generally, we tend to set apart pathogenic bacteria from opportunistic pathogens and commensal bacteria based on their abilities to induce disease in different hosts, however, the nature of the inflammatory response they induce in DC that set them apart from commensal bacteria remains largely unclear. In the present study, we developed a system by which we were able to compare the bacteria-induced imprint of important regulatory proteins in DC to bacterial-encoded ligands. We observed that DC responded to six different bacteria in a phyla-specific manner giving rise to similar inflammatory signatures within the groups of proteobacteria, firmicutes and actinobacteria, hence being independent on pathogenic versus non-pathogenic properties, and also on the bacteria-to-cell ratio for most bacteria. The results presented in this thesis add to the current knowledge about innate immunity to Salmonella, suggest new host immune cell subsets important for bacterial containment and provide a basic understanding of bacteria-induced DC inflammatory programs. The two latter could prove important in regard to treatment regimes, as targeted modulation of DC profiles for instance by probiotics, could lead to improved therapy for a number of gut related diseases.

SØrensen, Rikke Brandt; Pedersen, Susanne Brix

2012-01-01

 
 
 
 
261

Monitoring the efficacy of steam and formaldehyde treatment of naturally Salmonella-infected layer houses  

DEFF Research Database (Denmark)

Aims: To monitor if a temperature-humidity-time treatment found to be effective in eliminating Salmonella in laboratory trials (Gradel et al. 2003) was efficient against Salmonella in naturally infected layer houses. Methods and Results: Six layer houses with natural Salmonella infections were steam treated in a download period, aiming at greater than or equal to60degreesC and 100% relative humidity (RH) during a 24-h period, with or without the addition of 30 ppm formaldehyde. In addition, two control layer houses were disinfected chemically. Salmonella samples taken from predetermined sites before and after treatment were tested qualitatively for Salmonella and coliforms. Samples with indicator bacteria (feed inoculated with Escherichia coli or Enterococcus faecalis and faeces with naturally occurring E. coli and enterococci) were placed during steam-treatment at 12 sites in each house (where the temperature was logged at 5-min intervals) and tested for surviving bacteria. Generally, the field test results confirmed the results of laboratory tests, especially when 30 ppm formaldehyde was added to the steam. In well-sealed houses, the recommended temperature-humidity-time scheme was accomplished at a minimum of 10 cm above floor level within 1 h. Conclusions: A steam treatment of greater than or equal to60degreesC and 100% RH during a 24-h period with the addition of 30 ppm formaldehyde at the beginning of the process is recommended for eliminating Salmonella from naturally infected poultry layer houses. Significance and Impact of the Study: A specific recommendation for the elimination of Salmonella in poultry houses can be given.

2004-01-01

262

Significance of the bacteriophage treatment schedule in reducing Salmonella colonization of poultry.  

Science.gov (United States)

Salmonella remains the major cause of food-borne diseases worldwide, with chickens known to be the main reservoir for this zoonotic pathogen. Among the many approaches to reducing Salmonella colonization of broilers, bacteriophage offers several advantages. In this study, three bacteriophages (UAB_Phi20, UAB_Phi78, and UAB_Phi87) obtained from our collection that exhibited a broad host range against Salmonella enterica serovar Enteritidis and Salmonella enterica serovar Typhimurium were characterized with respect to morphology, genome size, and restriction patterns. A cocktail composed of the three bacteriophages was more effective in promoting the lysis of S. Enteritidis and S. Typhimurium cultures than any of the three bacteriophages alone. In addition, the cocktail was able to lyse the Salmonella enterica serovars Virchow, Hadar, and Infantis. The effectiveness of the bacteriophage cocktail in reducing the concentration of S. Typhimurium was tested in two animal models using different treatment schedules. In the mouse model, 50% survival was obtained when the cocktail was administered simultaneously with bacterial infection and again at 6, 24, and 30 h postinfection. Likewise, in the White Leghorn chicken specific-pathogen-free (SPF) model, the best results, defined as a reduction of Salmonella concentration in the chicken cecum, were obtained when the bacteriophage cocktail was administered 1 day before or just after bacterial infection and then again on different days postinfection. Our results show that frequent treatment of the chickens with bacteriophage, and especially prior to colonization of the intestinal tract by Salmonella, is required to achieve effective bacterial reduction over time. PMID:22773654

Bardina, Carlota; Spricigo, Denis A; Cortés, Pilar; Llagostera, Montserrat

2012-09-01

263

Molecular Typing of Salmonella paratyphi B and Salmonella paratyphi C Isolates from Clinical Samples in Iran  

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Full Text Available Background & Objective: Molecular typing is an important tool in surveillance and outbreak investigations of human Salmonella infections. In this study, Subtyping of Salmonella Paratyphi B and C isolates derived from Iranian patients was carried out by RAPD-PCR to assess the extent of genetic diversity of these isolates. Materials & Methods: Fourteen Salmonella isolates including 6 strains of Salmonella paratyphi B and 8 strains of Salmonella paratyphi C were characterized using RAPD-PCR. Two arbitrary primers, namely OPP-16 and P1254 were used for RAPD analysis and the dendrograms were constructed with NTsys 2.0 computer software. Results: Both primers showed high discriminatory power in differentiating of the related strains of Salmonella. The dendrograms constructed based on RAPD-PCR profiles (with both primers involving 14 salmonella strains revealed 4 distinct patterns, indicating that these isolates are genetically heterogeneous. Furthermore, a good correlation was not observed between the serotype and the molecular profiles obtained from RAPD data of the Salmonella isolates. Conclusion: The findings of the present study verify the usefulness of RAPD-PCR in characterizing and comparing strains of Salmonella Paratyphi B and C.

Fahimeh Baghbani-arani

2012-06-01

264

Assessment of attenuated Salmonella vaccine strains in controlling experimental Salmonella Typhimurium infection in chickens.  

Science.gov (United States)

Salmonella hold considerable promise as vaccine delivery vectors for heterologous antigens in chickens. Such vaccines have the potential additional benefit of also controlling Salmonella infection in immunized birds. As a way of selecting attenuated strains with optimal immunogenic potential as antigen delivery vectors, this study screened 20 novel Salmonella Typhimurium vaccine strains, differing in mutations associated with delayed antigen synthesis and delayed attenuation, for their efficacy in controlling colonization by virulent Salmonella Typhimurium, as well as for their persistence in the intestine and the spleen. Marked differences were observed between strains in these characteristics, which provide the basis for selection for further study as vaccine vectors. PMID:24396177

Pei, Yanlong; Parreira, Valeria R; Roland, Kenneth L; Curtiss, Roy; Prescott, John F

2014-01-01

265

Wall Geckos (Geckonidae) as reservoirs of Salmonellae in Nigeria: problems for epidemiology and public health.  

Science.gov (United States)

An investigation was carried out to establish the occurrence of Salmonellae in household wall lizards (Gecko gecko and Hemidactylus sp.) in Nsukka, Nigeria. Twentyseven out of 90 geckos examined yielded positive Salmonella isolations, giving a carrier-rate of 30 per cent. While 2 isolates were non-typable, the other 25 strains belonged to 6 serotypes: Salmonella weltevreden (8 strains), Salmonella typhimurium, Salmonella enteritidis, Salmonella hvittingfos, Salmonella saintpaul, and Salmonella agama. Two serotypes, Salmonella weltevreden and Salmonella hvittingfoss are reported for the first time in Nigeria. The role of Geckonidae in the epidemiology of salmonellosis and the public health implications of geckoborn Salmonellae are highlighted. PMID:3833829

Oboegbulem, S I; Iseghohimhen, A U

1985-09-01

266

EFSA Panel on Biological Hazards (BIOHAZ); Scientific Opinion on a review on the European Union Summary reports on trends and sources zoonoses, zoonotic agents and food-borne outbreaks in 2009 and 2010 – specifically for the data on Salmonella, Campylobacter, verotoxigenic Escherichia coli, Listeria monocytogenes and foodborne outbreaks  

DEFF Research Database (Denmark)

The European Union (EU) Summary Reports on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2009 and 2010 – specifically for the data on Salmonella, Campylobacter, verotoxigenic Escherichia coli, Listeria monocytogenes and foodborne outbreaks was reviewed. The main conclusions and recommendations are reported. Comparison between EU Member States (MSs) was found to be difficult due to the differences of the methods used, sampling schemes and reporting systems. Methods, sampling schemes and reporting systems among MSs should therefore be harmonised. When comparing MS-specific trends, the impact of sample sizes, weight of samples and methodologies should be considered, as these variables could otherwise lead to misinterpretation of the data. Incidence data alone do not provide a full picture of the public health burden of zoonotic diseases. Fatalities provide another important insight. Ultimately, summary measures of public health such as disability adjusted life years (DALYs) and cost-of-illness estimates should be presented. Travel information was found to be still incomplete in many MSs. For many pathogens this hampers source attribution. To better understand the public health problems related to food and animal sources in the EU, it is desirable to differentiate between travel within and outside the EU. This would also be useful to better evaluate the public health impact of EU-wide food safety measures. Whenever possible the data/results should be analysed using proper statistical tools. When data do not allow for this, the text should be kept to presenting the data without implying any patterns or trends.

2012-01-01

267

Effect of natural microbiota on growth of Salmonella spp. in fresh pork – A predictive microbiology approach  

DEFF Research Database (Denmark)

This study was undertaken to model and predict growth of Salmonella and the dominating natural microbiota, and their interaction in ground pork. Growth of Salmonella in sterile ground pork at constant temperatures between 4 °C and 38 °C was quantified and used for developing predictive models for lag time, max. specific growth rate and max. population density. Data from literature were used to develop growth models for the natural pork microbiota. Challenge tests at temperatures from 9.4 to 24.1 °C and with Salmonella inoculated in ground pork were used for evaluation of interaction models. The existing Jameson-effect and Lotka–Volterra species interaction models and a new expanded Jameson-effect model were evaluated. F-test indicated lack-of-fit for the classical Jameson-effect model at all of the tested temperatures and at 14.1–20.2 °C this was caused by continued growth of Salmonella after the natural microbiota had reached their max. population density. The new expanded Jameson-effect model and the Lotka–Volterra model performed better and appropriately described the continued but reduced growth of Salmonella after the natural microbiota had reached their max. population density. The expanded Jameson-effect model is a new and simple species interaction model, which performed as well as the more complex Lotka–Volterra model.

MØller, Cleide; Ilg, Y.

2013-01-01

268

Genetic parameters for resistance to the Salmonella abortusovis vaccinal strain Rv6 in sheep  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract An experimental population (1216 lambs from 30 sires of the Inra401 sheep was created in an Inra flock to allow QTL detection for susceptibility to Salmonella infection, wool and carcass traits. The Inra401 is a sheep composite line developed from two breeds: Berrichon du Cher and Romanov. At 113 days of age on average, the lambs were inoculated intravenously with 108 Salmonella abortusovis Rv6 (vaccinal strain. They were slaughtered 10 days after the inoculation. Several traits were measured at inoculation and/or slaughtering to estimate the genetic resistance of the lambs to Salmonella infection: specific IgM and IgG1 antibody titres, body weight loss, spleen and pre-scapular node weights and counts of viable Salmonella persisting in these organs. This paper presents a quantitative analysis of the genetic variability of the traits related to salmonellosis susceptibility. The heritabilities of the traits varied between 0.10 and 0.64 (significantly different from zero. Thus, in sheep as well as in other species, the determinism of resistance to Salmonella infection is under genetic control. Moreover, the correlations between the traits are in agreement with the known immune mechanisms. The genetic variability observed should help QTL detection.

Bouix Jacques

2003-03-01

269

Outbreak-associated Salmonella enterica serotypes and food Commodities, United States, 1998-2008.  

Science.gov (United States)

Salmonella enterica infections are transmitted not only by animal-derived foods but also by vegetables, fruits, and other plant products. To clarify links between Salmonella serotypes and specific foods, we examined the diversity and predominance of food commodities implicated in outbreaks of salmonellosis during 1998-2008. More than 80% of outbreaks caused by serotypes Enteritidis, Heidelberg, and Hadar were attributed to eggs or poultry, whereas >50% of outbreaks caused by serotypes Javiana, Litchfield, Mbandaka, Muenchen, Poona, and Senftenberg were attributed to plant commodities. Serotypes Typhimurium and Newport were associated with a wide variety of food commodities. Knowledge about these associations can help guide outbreak investigations and control measures. PMID:23876503

Jackson, Brendan R; Griffin, Patricia M; Cole, Dana; Walsh, Kelly A; Chai, Shua J

2013-08-01

270

Pyruvate kinase deficiency confers susceptibility to Salmonella typhimurium infection in mice  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The mouse response to acute Salmonella typhimurium infection is complex, and it is under the influence of several genes, as well as environmental factors. In a previous study, we identified two novel Salmonella susceptibility loci, Ity4 and Ity5, in a (AcB61 × 129S6)F2 cross. The peak logarithm of odds score associated with Ity4 maps to the region of the liver and red blood cell (RBC)–specific pyruvate kinase (Pklr) gene, which was previously shown to be mutated in AcB61. During Plasmodium...

Roy, Marie-france; Riendeau, Noe?mie; Be?dard, Christian; He?lie, Pierre; Min-oo, Gundula; Turcotte, Karine; Gros, Philippe; Canonne-hergaux, Franc?ois; Malo, Danielle

2007-01-01

271

Radiosensitivity study of salmonella enteritidis in chickens  

International Nuclear Information System (INIS)

One of the applications of ionizing radiations in food is the inactivation of vegetative phatogenic bacteria (radicidation) such as Salmonella, Shigella, Campylobacter, Vibro and Listeria. These bacteria are associated with the diseases transmitted by food (ETA). Fresh and frozen farmyard fowls can be contaminated with pathogenic microorganisms, between them Salmonella. In Argentine, between years 1987-1990, Salmonella enteritidis was the main cause of salmonellosis. In food irradiation, with the aim of improving and assuring its hygienic quality, it is important to know the radiosensitivity of microorganisms to be inactivated. Inactivation of a determined microorganism shall depend, between others factors, of the species, strain, number and of the irradiation conditions (temperature, media, etc.). D10 value is a very useful data in order to compare radiosensitivities between the microorganisms and the influence of different factors in their sensitivities. In this paper, it was determined the sensitivity to the gamma radiation of Salmonella enteritidis in fresh and frozen chickens

272

75 FR 18751 - Prevention of Salmonella  

Science.gov (United States)

...HEALTH AND HUMAN SERVICES Food and Drug Administration 21 CFR Part 118 [Docket No. FDA-2000-N-0190] Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation; Change of Registration Date, Address, and...

2010-04-13

273

Pet Turtles: Cute but Contaminated with Salmonella  

Science.gov (United States)

... Tobacco Products Vaccines, Blood & Biologics Articulos en Espanol Pet Turtles: Cute But Contaminated with Salmonella Search the ... infections from these animals still occur because some pet shops, flea markets, street vendors, and online stores ...

274

ANALYZING BIOSOLIDS FOR FECAL COLIFORM AND SALMONELLAE  

Science.gov (United States)

Current federal regulations required monitoring for fecal coliforms or Salmonella in biosolids destined for land application. Standard protocols designed to quantify these organisms in water or wastewater were identified and specified in these regulations. However, proto...

275

Rational design of Salmonella recombinant vaccines.  

Science.gov (United States)

Salmonella enterica is an important pathogen of animals and humans causing a variety of infectious diseases. The large number of cases of typhoid fever due to S. enterica serovar Typhi infections gives rise to the continuous need for improved vaccines against this life-threatening infection. However, S. enterica is also an interesting organism to act as a live attenuated carrier for the presentation of recombinant heterologous antigens. Comprehensive experimental studies have been performed and a detailed knowledge of the molecular mechanisms of important virulence factors is available. This allows the rationale design of improved Salmonella carrier strains and the development of novel strategies for the expression and presentation of recombinant antigens. Here, we review recent advances in generation of live attenuated Salmonella vaccines and discuss criteria for expression strategies of heterologous antigens by Salmonella carrier strains. PMID:17888730

Cheminay, Cédric; Hensel, Michael

2008-01-01

276

Water Frogs, Aquariums, and Salmonella -- Oh My!  

Centers for Disease Control (CDC) Podcasts

This CDC Kidtastics podcast discusses how people can get Salmonella from water frogs and aquariums.  Created: 12/9/2009 by National Center for Zoonotic, Vector-Borne, and Enteric Diseases (NCZVED).   Date Released: 12/9/2009.

2009-12-09

277

DETECTION OF FRNA COLIPHAGES IN GROUNDWATER: INTERFERENCE WITH THE ASSAY BY SOMATIC SALMONELLA BACTERIOPHAGES  

Science.gov (United States)

Groundwater samples from two sites in Alabama, USA were plaque assayed for F-specific RNA (FRNA) coliphages using Salmonella typhimurium WG49 as the host bacterium. While numerous plaques were detected with WG49 (a strain possessing Escherichia coli F pili), plaques were also obs...

278

Study of 163 children with invasive Salmonella infection in pediatric medical center1  

Directory of Open Access Journals (Sweden)

Full Text Available Invasive salmonellosis is common in tropical areas. This study examines the performance of a clinical definition for its recognition among children ages 1 to 14 years admitting to a referral pediatric hospital in Tehran. 60 children were enrolled into the study during a period of 51 months. To facilitate analysis, cases were divided into 5 categories according to the likelihood of invasive salmonellosis with category A representing microbiologically confirmed salmonella bacteremia 17 (28.3% and 6 (10% with positive bone marrow cultures. And category D representing those cases in which an alternative diagnosis was firmly established. Salmonella serology supported invasive salmonellosis as the diagnosis in 17 (28% of the nonbacteremic children (category B and C. Salmonella serology suggested that invasive salmonellosis without detectable bacteremia was common. Blood culture proved and serologically diagnosed cases shows that the definition has a specificity of at least 60%.

Khotaeei Gh

2000-07-01

279

[Salmonella Freetown: 1st isolation in Argentina].  

Science.gov (United States)

In this paper we report the first case of Salmonella Freetown in Argentina. It was isolated from a stool sample of a child of 1 month and 7 days, assisted as outpatient in the Hospital de Niños Dr. Ricardo Gutiérrez of the city of Santa Fe. On the basis of the biochemical characteristics and antigenic formulae, this new serovar belongs to the species Salmonella enterica subspecies enterica. PMID:8768486

Caffer, M I; Mayoral, C; Bruno, S; Alcain, A; Terragno, R

1996-01-01

280

Cambios epidemiológicos de las salmonelosis en Chile: Desde Salmonella typhi a Salmonella enteritidis CHANGES IN EPIDEMIOLOGICAL PATTERNS OF SALMONELLOSIS IN CHILE: SINCE Salmonella typhi TO Salmonella enteritidis  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Chile ha experimentado un cambio epidemiológico en la última década con la desaparición progresiva de la fiebre tifoidea causada mayoritariamente por Salmonella typhi y la emergencia epidémica de Salmonella enteritidis, un agente de diarrea sin tratamiento específico eficaz y ligado estrechamente a productos avícolas contaminados e inadecuadamente preparados. La fiebre tifoidea ha disminuido su importancia debido al desarrollo humano experimentado en Chile que ha significado un alto gr...

Alberto Fica, C.; Marcela Alexandre, S.; Soledad Prat, M.; Alda Ferna?ndez, R.; Jorge Ferna?ndez, O.; Ingrid Heitmann, G.

2001-01-01

 
 
 
 
281

Deciphering interplay between Salmonella invasion effectors.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Bacterial pathogens have evolved a specialized type III secretion system (T3SS) to translocate virulence effector proteins directly into eukaryotic target cells. Salmonellae deploy effectors that trigger localized actin reorganization to force their own entry into non-phagocytic host cells. Six effectors (SipC, SipA, SopE/2, SopB, SptP) can individually manipulate actin dynamics at the plasma membrane, which acts as a 'signaling hub' during Salmonella invasion. The extent of crosstalk between...

2008-01-01

282

Rapid detection of Salmonella in foods using a combination of SPRINT TM,MSRV TM and Salmonella Latex TestTM Detecção rápida de Salmonella em alimentos empregando uma combinação de SPRINT®, MSRV® e Salmonella Latex Test®  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A new procedure for rapid detection of Salmonella in foods, based on the combination of SPRINT TM, MSRV TM and Salmonella Latex TestTM, was evaluated. SPRINT TM is a system to reduce the preenrichment and selective enrichment steps to 24 hours. MSRV TM is a semi-solid selective media for detection of motile Salmonella. Salmonella Latex TestTM is a rapid latex agglutination test for Salmonella. Using the three systems in combination, the total time for detection of Salmonella in a food sample ...

Jane Maria Lafayette Neves Gelinski; Gunnar Martin; Maria Teresa Destro; Mariza Landgraf; Bernadette Dora Gombossy de Melo Franco

2002-01-01

283

Sensibilidad a los antimicrobianos de cepas de salmonella aisladas en granjas porcinas del estado Zulia / Antimicrobial susceptibility of salmonella strains isolated from pig herds in Zulia state  

Scientific Electronic Library Online (English)

Full Text Available SciELO Venezuela | Language: Spanish Abstract in spanish El objetivo de este estudio fue determinar los patrones de resistencia a los antimicrobianos de diferentes cepas de Salmonella aisladas en granjas de cerdos del estado Zulia. Para este fin se evaluaron mediante la técnica de Bauer-Kirby, 126 cepas de Salmonella aisladas de heces de cerdos portadores [...] asintomáticos. Las pruebas de sensibilidad antimicrobiana demostraron que los más altos niveles de resistencia fueron frente a la sulfamida (54%), tetraciclina (40%), ácido nalidíxico (29%) y ampicilina (23%). Sin embargo, sensibilidad superior al 95% fue encontrada frente a la ceftriaxona, gentamicina, apramicina y colistina. El treinta por ciento de las cepas mostraron multirresistencia (MR) a los antimicrobianos, siendo el patrón de resistencia ASuT (7,14%) el más frecuente. Los resultados obtenidos indican que la proporción de cepas de Salmonella de origen porcino con características de multirresistencia a los agentes antimicrobianos es medianamente elevada (30%) y esta multirresistencia puede afectar a cualquier serotipo. Desde ese punto de vista, la infección de las personas por cepas de Salmonella de origen porcino conlleva a un riesgo potencial de presentar dificultades en el tratamiento específico. Abstract in english The aim of this study was to determine antimicrobial resintance paterns of different strains of Salmonella isolated in pig farms of the Zulia State. To achieve these goals 126 strain Salmonella were screened by Kirby-Bauer method, colleted from heces of pigs asymptomatic. Antimicrobial susceptibilit [...] y tests showed that the highest level of resistance was against Sulphonamides (54%), Tetracycline (40%), Nalidixic acid (29%) and Amplicillin (23%). However, susceptibility superior to 95% was found to Ceftriaxone, Gentamycin, Apramycin and Colistin. Thrity percent of the strains showed multirresitance, being the patterns resistance ASuT (7.14%) the most frequent. The results indicate the proportion of strain of Salmonella of pig origin with characteristics of multiresistance to the antimicrobial agents is elevated (30%) and this multiresistance could affect to anyone serotype. From this point of view, the infection of the people by isolates of Salmonella from swine origin entails a potential risk to present difficulties in the specific treatment.

Willian, Mejia; Derwin, Calatayud Marquez; Denice, Zapata; Armando, Quintero; Damarys, Sánchez; Enric, Mateu.

2008-12-01

284

SALMONELLA ENTERITIDIS – PHENOTYPIC AND GENOTYPIC  

Directory of Open Access Journals (Sweden)

Full Text Available Today, Salmonella enterica subspecies enterica serovar Enteritidis (S. Enteritidis represents one of the most common serotypes that causes enterocolitis. Since S. enteritidis identification methods are advanced permanently, the following phenotyping methods could be applied for this purpose: biotyping, phagotyping (phage typing – PT, and resistotyping. From methods for genotyping of S. Enteritidis, plasmid profile analysis (PP, restriction analysis of the virulence plasmid, ribotyping, pulsed field gel electrophoresis (PFGE, insertion sequences, polymerase chain reaction (PCR, random amplified polymorphic DNA analysis (RAPD could be applied. On the one hand, S. Enteritidis expresses clearly homogenous structure which is reflected by domination of few phagotypes, presence of one plasmid profile in most of strains, merely three clonal lines, as well as a large number of electrophoretic types in a single dendrogram line. Insufficient discrimination of typing systems of S. Enteritidis suggests the introduction of new typing methods as well as improvement of the old ones.

Biljana Miljkovi?-Selimovi?

2009-10-01

285

A real-time PCR method for the detection of Salmonella enterica from food using a target sequence identified by comparative genomic analysis.  

Science.gov (United States)

A 5'-nuclease real-time PCR assay using a minor groove binding probe was developed for the detection of Salmonella enterica from artificially contaminated foods. S. enterica-specific sequences were identified by a comparative genomic approach. Several species-specific target sequences were evaluated for specificity. A real-time PCR assay was developed targeting a nucleotide sequence within the putative type III secretion ATP synthase gene (ssaN). An internal amplification control (IAC) probe was designed by randomly shuffling the target probe sequence and a single-stranded oligonucleotide was synthesized to serve as an IAC. The assay demonstrated 100% inclusivity for the 40 Salmonella strains tested and 100% exclusivity for 24 non-Salmonella strains. The detection limit of the real-time PCR assay was 41.2 fg/PCR with Salmonella Typhimurium genomic DNA and 18.6 fg/PCR using Salmonella Enteritidis genomic DNA; 8 and 4 genome equivalents, respectively. In the presence of a natural background flora derived from chicken meat enrichment cultures, the sample preparation and PCR method were capable of detecting as few as 130 Salmonella cfu/mL. Using the developed real-time PCR method to detect Salmonella in artificially contaminated chicken, liquid egg and peanut butter samples, as few as 1 cfu/10 g of sample was detectable after a brief (6h) non-selective culture enrichment. PMID:20060189

Chen, Jing; Zhang, Lida; Paoli, George C; Shi, Chunlei; Tu, Shu-I; Shi, Xianming

2010-02-28

286

Evaluation of selective enrichment broths and chromogenic media for Salmonella detection in highly contaminated chicken carcasses.  

Science.gov (United States)

We evaluated the effectiveness of 2 selective enrichment broths, Rappaport-Vassiliadis Soy (RVS) and Muller-Kauffmann tetrathionate with novobiocin (MKTTn), for the isolation of Salmonella from chicken carcasses obtained from 3 different types of retail markets. We also compared a chromogenic agar, chromID Salmonella agar (SM-ID 2), with a classic plating medium, xylose lysine deoxycholate agar (XLD). Salmonella were isolated from 118 of the 180 samples (65.5%). Salmonella were detected in 105 samples (88%) plated on XLD and 111 samples (94%) plated on SM-ID 2 when RVS broth was used for enrichment, and 43 samples (36.4%) plated on XLD and 67 samples (56.8%) plated on SM-ID 2 when the MKTTn broth was used. The highest sensitivity was found in the RVS-XLD combination (0.99), followed by RVS-SM-ID 2 (0.97). The specificity of the RVS-SM-ID 2 combination was the highest (0.89), but that of the MKTTn-XLD combination was zero. The results of this study indicate that the selective enrichment broths had a great effect on the sensitivity and specificity of plating media, and our study confirms that the RVS broth is the most suitable enrichment for the investigation of Salmonella in chicken carcasses. This observation suggests that use of RVS broth for selective enrichment and SM-ID 2 for selective isolation may be the best combination to determine the presence of Salmonella in chicken carcasses. PMID:22499882

Hyeon, J-Y; Park, J-H; Chon, J-W; Wee, S-H; Moon, J-S; Kim, Y-J; Seo, K-H

2012-05-01

287

A comparative study of culture methods and PCR assay for Salmonella detection in poultry drinking water.  

Science.gov (United States)

The present work compared 2 culture methods and PCR assays for motile and nonmotile Salmonella detection using artificially contaminated poultry drinking water. The specificity was 1 for all methods studied. The accuracy and sensitivity were 1 for all motile strains, whereas these parameters were between 0 and 0.7 for nonmotile Salmonella strains. The positive predictive value and negative predictive value were 1 for all motile Salmonella strains in the 3 methods used. Nonmotile Salmonella strains showed a positive predictive value of 1 in the PCR method. However, the positive predictive value was indeterminate in the tetrathionate (TT) methods for both strains tested and in the modified semisolid Rappaport-Vassiliadis (MSRV) method for Salmonella Pullorum. On the other hand, the negative predictive value was between 0.20 and 0.43 for the 3 methods. The detection level of motile strains was 4 to 7 cfu/25 mL for all methods. Nonmotile Salmonella strains could not be detected in the TT method, whereas only Salmonella Gallinarum could be recovered from 1.1 × 10(1) cfu/25 mL in the MSRV method. In relation to the molecular methods, PCR could detect these strains from 1.1 × 10(4) cfu/25 mL. Extending incubation time of the enrichment medium to 6 d in the TT method did not improve the isolation rates. In general, all selective plating media did not show any statistical differences in the parameters of performance studied. The kappa coefficient showed that there was an excellent agreement between the 3 methods for motile strains. For nonmotile strains, the agreement was poor between the MSRV and the PCR; there was no agreement when the TT method was compared with the MSRV and the PCR methods. The difference in detection levels obtained with the methods used for motile and nonmotile Salmonella strains and the difficulty in detecting these last strains represents a potential problem when a poultry water sample is considered negative for the presence of Salmonella. PMID:23243252

Soria, M C; Soria, M A; Bueno, D J

2013-01-01

288

The relationship between the numbers of Salmonella Enteritidis, Salmonella Heidelberg, or Salmonella Hadar colonizing reproductive tissues of experimentally infected laying hens and deposition inside eggs.  

Science.gov (United States)

Contamination of eggs by Salmonella Enteritidis has been a prominent cause of human illness for several decades and is the focus of a recently implemented national regulatory plan for egg-producing flocks in the United States. Salmonella Heidelberg has also been identified as an egg-transmitted pathogen. The deposition of Salmonella strains inside eggs is a consequence of reproductive tract colonization in infected laying hens, but prior research has not determined the relationship between the numbers of Salmonella that colonize reproductive organs and the associated frequency of egg contamination. In the present study, groups of laying hens in two trials were experimentally infected with large oral doses of strains of Salmonella Enteritidis (phage type 13a), Salmonella Heidelberg, or Salmonella Hadar. Reproductive tissues of selected hens were cultured to detect and enumerate Salmonella at 5 days postinoculation, and the interior contents of eggs laid between 6 and 25 days postinoculation were tested for contamination. Significantly more internally contaminated eggs were laid by hens infected with Salmonella Enteritidis (3.58%) than with strains of either Salmonella Heidelberg (0.47%) or Salmonella Hadar (0%). However, no significant differences were observed between Salmonella strains in either isolation frequency or the number of colony-forming units (CFU) isolated from ovaries or oviducts. Salmonella isolation frequencies ranged from 20.8% to 41.7% for ovaries and from 8.3% to 33.3% for oviducts. Mean Salmonella colonization levels ranged from 0.10 to 0.51 log CFU/g for ovaries and from 0.25 to 0.46 log CFU/g for oviducts. Although parallel rank-orders were observed for Salmonella enumeration (in both ovaries and oviducts) and egg contamination frequency, a statistically significant relationship could not be established between these two parameters of infection. PMID:21793440

Gast, Richard K; Guraya, Rupa; Guard, Jean; Holt, Peter S

2011-06-01

289

SALMONELLA REGROWTH IN COMPOST AS INFLUENCED BY SUBSTRATE (SALMONELLA REGROWTH IN COMPOST)  

Science.gov (United States)

Composting can eliminate pathogenic organisms, including salmonellae, from sewage sludge. However, if salmonellae are present in the compost at undetectable levels or are inoculated into the compost by infected animals or from other sources, they may regrow presenting a health ha...

290

Autophagy facilitates Salmonella replication in HeLa cells.  

Science.gov (United States)

Autophagy is a process whereby a double-membrane structure (autophagosome) engulfs unnecessary cytosolic proteins, organelles, and invading pathogens and delivers them to the lysosome for degradation. We examined the fate of cytosolic Salmonella targeted by autophagy and found that autophagy-targeted Salmonella present in the cytosol of HeLa cells correlates with intracellular bacterial replication. Real-time analyses revealed that a subset of cytosolic Salmonella extensively associates with autophagy components p62 and/or LC3 and replicates quickly, whereas intravacuolar Salmonella shows no or very limited association with p62 or LC3 and replicates much more slowly. Replication of cytosolic Salmonella in HeLa cells is significantly decreased when autophagy components are depleted. Eventually, hyperreplication of cytosolic Salmonella potentiates cell detachment, facilitating the dissemination of Salmonella to neighboring cells. We propose that Salmonella benefits from autophagy for its cytosolic replication in HeLa cells. IMPORTANCE As a host defense system, autophagy is known to target a population of Salmonella for degradation and hence restricting Salmonella replication. In contrast to this concept, a recent report showed that knockdown of Rab1, a GTPase required for autophagy of Salmonella, decreases Salmonella replication in HeLa cells. Here, we have reexamined the fate of Salmonella targeted by autophagy by various cell biology-based assays. We found that the association of autophagy components with cytosolic Salmonella increases shortly after initiation of intracellular bacterial replication. Furthermore, through a live-cell imaging method, a subset of cytosolic Salmonella was found to be extensively associated with autophagy components p62 and/or LC3, and they replicated quickly. Most importantly, depletion of autophagy components significantly reduced the replication of cytosolic Salmonella in HeLa cells. Hence, in contrast to previous reports, we propose that autophagy facilitates Salmonella replication in the cytosol of HeLa cells. PMID:24618251

Yu, Hong B; Croxen, Matthew A; Marchiando, Amanda M; Ferreira, Rosana B R; Cadwell, Ken; Foster, Leonard J; Finlay, B Brett

2014-01-01

291

Application of Real-time PCR for Rapid Petection of Salmonella spp., Salmonella enterica ser. Typhimurium and Enteritidis in Food Samples of Animal Origin without Pre-enrichment and with Pre-enrichment  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study was follow the contamination of milk and meat products with Salmonella spp. by using the Step One real-time PCR. We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and Power SYBR Green PCR Master Mix for pursuance the real-time PCR (Applied Biosystems. We were observing the presence of genes stn (Salmonella enterica ser. Typhimurium DT096, sef and pef (Salmonella enterica ser. Enteritidis SE7. We could detect strain of Salmonella spp. in the investigated samples without pre-enrichment and after pre-enrichment. Our results indicated that the Step One real-time PCR assay developed in this study could sensitively detect Salmonella spp. in food samples of animal origin (swabs. This Step One real-time PCR assay is extremely useful for any laboratory in possession of a real-time PCR. It is a fast, reproducible, simple, specific and sensitive way to detect nucleic acids, which could be used in clinical diagnostic tests in the future. Our results indicated that the Step One real-time PCR assay developed in this study could sensitively detect Salmonella spp. in ready to eat food. Thus, these results proved real-time PCR to be useful as a rapid diagnostic test for the direct detection of pathogens in food of animal origin, without the need of pre-enrichment steps.

Jaroslav Pochop

2012-05-01

292

European Food Safety Authority; Analysis of the baseline survey of Salmonella in holdings with breeding pigs, in the EU, 2008; Part B: Analysis of factors potentially associated with Salmonella pen positivity  

DEFF Research Database (Denmark)

A European Union-wide Salmonella baseline survey was conducted in 2008 in holdings with breeding pigs. A total of 1,609 randomly selected holdings housing and selling mainly breeding pigs (breeding holdings) and 3,508 holdings housing commercial breeding pigs and mainly selling pigs for fattening or slaughter (production holdings) were sampled. In each selected holding, pooled fresh faecal samples were collected from 10 randomly chosen pens of breeding pigs over six months of age, representing the different stages of the breeding herd, and examined for the presence of Salmonella. Analyses at country-level demonstrated a strong positive association between the prevalence of Salmonella-positive breeding holdings and the prevalence of Salmonella-positive production holdings, suggesting a vertical dissemination of Salmonella between the holdings. Based on the combined results from breeding and production holdings, multivariable regression analysis showed that the odds of Salmonella-positive pens with pigs increased with the number of breeding pigs in the holding and with the following pen-level factors: flooring systems other than slatted floors or solid floors with straw, presence of maiden gilts, number of pigs per pen, feed of commercial compound origin or pelleted feed. A tendency towards some Member State group-specific Salmonella serovars was identified, but spatial distribution of other serovars was heterogeneous. S. Typhimurium and S. Derby were widespread and dominant in the EU, in both breeding and production holdings. However, many other serovars were relatively prevalent in Western EU Member States. A complementary within-holding prevalence study indicated that, due to a non-perfect diagnostic test sensitivity, the observed EU-level prevalence of Salmonella-positive holdings with breeding pigs was roughly 80% of the estimated true EU-level prevalence. But this proportion varied between Member States.

2011-01-01

293

The sensitivity of real-time PCR amplification targeting invasive Salmonella serovars in biological specimens  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background PCR amplification for the detection of pathogens in biological material is generally considered a rapid and informative diagnostic technique. Invasive Salmonella serovars, which cause enteric fever, can be commonly cultured from the blood of infected patients. Yet, the isolation of invasive Salmonella serovars from blood is protracted and potentially insensitive. Methods We developed and optimised a novel multiplex three colour real-time PCR assay to detect specific target sequences in the genomes of Salmonella serovars Typhi and Paratyphi A. We performed the assay on DNA extracted from blood and bone marrow samples from culture positive and negative enteric fever patients. Results The assay was validated and demonstrated a high level of specificity and reproducibility under experimental conditions. All bone marrow samples tested positive for Salmonella, however, the sensitivity on blood samples was limited. The assay demonstrated an overall specificity of 100% (75/75 and sensitivity of 53.9% (69/128 on all biological samples. We then tested the PCR detection limit by performing bacterial counts after inoculation into blood culture bottles. Conclusions Our findings corroborate previous clinical findings, whereby the bacterial load of S. Typhi in peripheral blood is low, often below detection by culture and, consequently, below detection by PCR. Whilst the assay may be utilised for environmental sampling or on differing biological samples, our data suggest that PCR performed directly on blood samples may be an unsuitable methodology and a potentially unachievable target for the routine diagnosis of enteric fever.

Chau Tran

2010-05-01

294

Salmonella enterica Serovar Enteritidis Enterocolitis during Late Stages of Gestation Induces an Adverse Pregnancy Outcome in the Murine Model  

Science.gov (United States)

Foodborne diseases caused by Salmonella enterica serovar Enteritidis (S. Enteritidis) are a significant health problem. Pregnancy, state of immunological tolerance, is a predisposing condition for the development of infections with intracellular pathogens. Salmonella species can cause pregnancy complications such as chorioamnionitis, transplacental fetal infection, pre term labor, abortions, neonatal and maternal septicemia. However, the specific mechanisms by which Salmonella infections trigger these alterations are not clear. In the present work, using a self-limiting enterocolitis murine model, we show that the ingestion of a low dose of S. Enteritidis at late stages of pregnancy (day 15 of gestation) is sufficient to induce massive maternal infection. We found that Salmonella infection leads to 40% of pre term delivery, 33% of abortion and fetal growth restriction. Placental dysfunction during S. Enteritidis enterocolitis was confirmed through cellular infiltration and hypoxia markers (MPO activity and COX-1 and COX-2 expression, respectively). Apoptosis in placental tissue due to Salmonella infection was also evident at day 18 of gestation when investigated by morphometric procedure, DNA fragmentation and Fas/FasL expression. Also, the expression of IFN-?, TNF-?, IL-17 and IL-10 was up regulated in response to Salmonella not only in placenta, but also in amniotic fluid and maternal serum. Altogether, our results demonstrate that S. Enteritidis enterocolitis during late stages of gestation causes detrimental effect on pregnancy outcome. PMID:25365504

Noto Llana, Mariangeles; Sarnacki, Sebastian Hernan; Aya Castaneda, Maria del Rosario; Pustovrh, Maria Carolina; Gartner, Alejandra Sonia; Buzzola, Fernanda Roxana; Cerquetti, Maria Cristina; Giacomodonato, Monica Nancy

2014-01-01

295

Salmonella enterica Serovar Enteritidis Enterocolitis during Late Stages of Gestation Induces an Adverse Pregnancy Outcome in the Murine Model.  

Science.gov (United States)

Foodborne diseases caused by Salmonella enterica serovar Enteritidis (S. Enteritidis) are a significant health problem. Pregnancy, state of immunological tolerance, is a predisposing condition for the development of infections with intracellular pathogens. Salmonella species can cause pregnancy complications such as chorioamnionitis, transplacental fetal infection, pre term labor, abortions, neonatal and maternal septicemia. However, the specific mechanisms by which Salmonella infections trigger these alterations are not clear. In the present work, using a self-limiting enterocolitis murine model, we show that the ingestion of a low dose of S. Enteritidis at late stages of pregnancy (day 15 of gestation) is sufficient to induce massive maternal infection. We found that Salmonella infection leads to 40% of pre term delivery, 33% of abortion and fetal growth restriction. Placental dysfunction during S. Enteritidis enterocolitis was confirmed through cellular infiltration and hypoxia markers (MPO activity and COX-1 and COX-2 expression, respectively). Apoptosis in placental tissue due to Salmonella infection was also evident at day 18 of gestation when investigated by morphometric procedure, DNA fragmentation and Fas/FasL expression. Also, the expression of IFN-?, TNF-?, IL-17 and IL-10 was up regulated in response to Salmonella not only in placenta, but also in amniotic fluid and maternal serum. Altogether, our results demonstrate that S. Enteritidis enterocolitis during late stages of gestation causes detrimental effect on pregnancy outcome. PMID:25365504

Noto Llana, Mariángeles; Sarnacki, Sebastián Hernán; Aya Castañeda, María Del Rosario; Pustovrh, María Carolina; Gartner, Alejandra Sonia; Buzzola, Fernanda Roxana; Cerquetti, María Cristina; Giacomodonato, Mónica Nancy

2014-01-01

296

Serologic reactions against Salmonella in samples from broiler parent stock with and without preceding colibacillosis: A case-control study  

DEFF Research Database (Denmark)

In the Danish Salmonella Control Program, eggs from broiler parent flocks are surveyed by serologic analysis every 4 wk for antibodies against Salmonella lipopolysaccharide O-antigens 1, 4, 5, 9, and 12 (Mix-enzyme-linked immunosorbent assay [ELISA]) and 6 and 7 (Infantis-ELISA). The antibody response is measured in percentage optical density (OD%) of a strong positive reaction, and the cutoff value has been determined to be 40 OD%. Two or more reactors above 40 OD% will place the parent flock under suspicion. There has been concern about possible cross-reactions between Salmonella spp. and other Enterobacteriaceae, e.g., Escherichia coli, because a high specificity of a Salmonella antibody test is desirable. Moreover, false-positive Salmonella results have economic consequences and impede planning the production. A case-control study based on cases of clinical E. coli infections (colibacillosis) from two Danish hatcheries, supplying about 62% of the Danish broiler production, is described. In order to eliminate a possible bias from age and season, the controls were matched on age of the birds and on time of submitting the samples. This study shows that flocks with preceding colibacillosis did nor have higher salmonella reactions than matched flocks without a preceding colibacillosis. This observation was confirmed in longitudinal studies.

Feld, Niels Christian

2001-01-01

297

Guide to National Control Programme for Salmonella in laying flocks  

Jan 1, 2009 ... In brief, the National Control Programme (NCP) for Salmonella in ... that, for the \\protection of human health, coherent action to reduce Salmonella serotypes \\considered ... The identification of the flock (building/flock identity).

298

21 CFR 118.4 - Salmonella Enteritidis (SE) prevention measures.  

Science.gov (United States)

...and Drugs 2 2010-04-01 2010-04-01 false Salmonella Enteritidis (SE) prevention measures. 118.4 Section...STORAGE, AND TRANSPORTATION OF SHELL EGGS § 118.4 Salmonella Enteritidis (SE) prevention measures. You...

2010-04-01

299

Tips to Reduce Your Risk of Salmonella from Eggs  

Science.gov (United States)

... CDC Features Tips to Reduce Your Risk of Salmonella from Eggs Language: English Español (Spanish) Share Compartir ... can take to reduce my risk of a Salmonella infection from eggs? Like other foods, keep eggs ...

300

21 CFR 118.4 - Salmonella Enteritidis (SE) prevention measures.  

Science.gov (United States)

...2010-04-01 2010-04-01 false Salmonella Enteritidis (SE) prevention measures...TRANSPORTATION OF SHELL EGGS § 118.4 Salmonella Enteritidis (SE) prevention measures...poultry, wild birds, cats, and other animals from entering poultry houses; and...

2010-04-01

 
 
 
 
301

78 FR 42451 - Animal Feeds Contaminated With Salmonella Microorganisms  

Science.gov (United States)

...Docket No. FDA-2013-N-0253] Animal Feeds Contaminated With Salmonella Microorganisms...revoking an advisory opinion on animal feeds contaminated with Salmonella microorganisms...byproducts intended for use in animal feed may be contaminated with...

2013-07-16

302

21 CFR 118.4 - Salmonella Enteritidis (SE) prevention measures.  

Science.gov (United States)

... false Salmonella Enteritidis (SE) prevention measures. 118.4 Section 118.4...118.4 Salmonella Enteritidis (SE) prevention measures. You must follow the SE prevention measures set forth in this section....

2010-04-01

303

40 CFR 79.68 - Salmonella typhimurium reverse mutation assay.  

Science.gov (United States)

...multiple base pairs in the DNA molecule. Salmonella...blue 1. (d) Test method —(1) Principle...Ames, B. N., Revised methods for the Salmonella mutagenicity...D. “Analysis of a method for testing azo dyes...Nitropyrenes: Isolation, identification,...

2010-07-01

304

Prevalence and characterization of motile Salmonella in commercial layer poultry farms in Bangladesh  

DEFF Research Database (Denmark)

Salmonella is a globally widespread food-borne pathogen having major impact on public health. All motile serovars of Salmonella enterica of poultry origin are zoonotic, and contaminated meat and raw eggs are an important source to human infections. Information on the prevalence of Salmonella at farm/holding level, and the zoonotic serovars circulating in layer poultry in the South and South-East Asian countries including Bangladesh, where small-scale commercial farms are predominant, is limited. To investigate the prevalence of Salmonella at layer farm level, and to identify the prevalent serovars we conducted a cross-sectional survey by randomly selecting 500 commercial layer poultry farms in Bangladesh. Faecal samples from the selected farms were collected following standard procedure, and examined for the presence of Salmonella using conventional bacteriological procedures. Thirty isolates were randomly selected, from the ninety obtained from the survey, for serotyping and characterized further by plasmid profiling and pulsed-field gel electrophoresis (PFGE). Results of the survey showed that the prevalence of motile Salmonella at layer farm level was 18% (95% confidence interval 15-21%), and Salmonella Kentucky was identified to be the only serovar circulating in the study population. Plasmid analysis of the S. Kentucky and non-serotyped isolates revealed two distinct profiles with a variation of two different sizes (2.7 and 4.8 kb). PFGE of the 30 S. Kentucky and 30 non-serotyped isolates showed that all of them were clonally related because only one genotype and three subtypes were determined based on the variation in two or three bands. This is also the first report on the presence of any specific serovar of Salmonella enterica in poultry in Bangladesh.

Barua, Himel; Biswas, Paritosh K.

2012-01-01

305

In vitro assessment of the susceptibility of planktonic and attached cells of foodborne pathogens to bacteriophage p22-mediated salmonella lysates.  

Science.gov (United States)

This study was designed to evaluate the lytic activity of bacteriophage P22 against Salmonella Typhimurium ATCC 19585 (Salmonella Typhimurium P22(-)) at various multiplicities of infections (MOIs), the susceptibility of preattached Salmonella cells against bacteriophage P22, and the effect of P22-mediated bacterial lysates (extracellular DNA) on the attachment ability of Listeria monocytogenes ATCC 7644 and enterohemorrhagic Escherichia coli ATCC 700927 to surfaces. The numbers of attached Salmonella Typhimurium P22(-) cells were effectively reduced to below the detection limit (1 log CFU/ml) at the fixed inoculum levels of 3 × 10(-) CFU/ml (MOI = 3.12) and 3 × 10(3) CFU/ml (MOI = 4.12) by bacteriophage P22. The attached Salmonella Typhimurium P22(-) cells remained more than 2 log CFU/ml, with increasing inoculum levels from 3 × 10(4) to 3 × 10(7) CFU/ml infected with 4 × 10(8) PFU/ml of P22. The number of preattached Salmonella Typhimurium P22(-) cells was noticeably reduced by 2.72 log in the presence of P22. The highest specific attachment ability values for Salmonella Typhimurium P22(-), Salmonella Typhimurium ATCC 23555 carrying P22 prophage (Salmonella Typhimurium P22(+)), L. monocytogenes, and enterohemorrhagic E. coli were 2.09, 1.06, 1.86, and 1.08, respectively, in the bacteriophage-mediated cell-free supernatants (CFS) containing high amounts of extracellular DNA. These results suggest that bacteriophages could potentially be used to effectively eliminate planktonic and preattached Salmonella Typhimurium P22(-) cells with increasing MOI. However, further research is needed to understand the role of bacteriophage-induced lysates in bacterial attachment, which can provide useful information for the therapeutic use of bacteriophage in the food system. PMID:24290682

Ahn, Juhee; Kim, Songrae; Jung, Lae-Seung; Biswas, Debabrata

2013-12-01

306

Verbeteren van Salmonella isolatie-methoden uit kippenfaeces  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Een van de taken van het Communautair Referentie Laboratorium voor Salmonella (CRL-Salmonella) is het verbeteren van detectie methoden voor Salmonella. Hiertoe werd onderzoek uitgevoerd om de isolatie van Salmonella uit kippenfaeces te verbeteren. In vergelijking met de standaard ISO methode werden de volgende effecten bestudeerd: (i) de invloed van de incubatietijd van de voorophoping in gebufferd pepton water (BPw); (ii) het gebruik van Modified Semi-Solid Rappaport Vassiliadis Medium Ba...

Cj, Heuvelman; Veld Ph, In Apos T.

2012-01-01

307

Salmonella enterica: a surprisingly well-adapted intracellular lifestyle  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The infectious intracellular lifestyle of Salmonella enterica relies on the adaptation to nutritional conditions within the Salmonella-containing vacuole (SCV) in host cells. We summarize latest results on metabolic requirements for Salmonella during infection. This includes intracellular phenotypes of mutant strains based on metabolic modeling and experimental tests, isotopolog profiling using 13C-compounds in intracellular Salmonella, and complementation of metabolic defects for attenuated ...

Dandekar, Thomas; Astrid, Fieselmann; Jasmin, Popp; Hensel, Michael

2012-01-01

308

Resistance of broiler outbred lines to infection with Salmonella enteritidis  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Salmonella infections originating from poultry are one of the major causes of food-borne disease. For the control of salmonella in poultry a multifactorial approach is more likely to be effective, and the genetic resistance of poultry breeds to salmonella infections may be a valuable contribution. Experimental Salmonella enteritidis infections were examined in three different broiler outbred lines: the FC line, which had been selected for feed conversion efficiency; the R line, which had been...

Bolder, N. M.; Janss, L. L. G.; Putirulan, F. F.; Wagenaar, J. A.

2002-01-01

309

Prevalence of Salmonella on chicken carcasses from retail markets in Vietnam.  

Science.gov (United States)

This study was conducted to estimate the prevalence of Salmonella on chicken carcasses collected from six regions in Vietnam. A total of 1,000 whole, dressed chicken carcasses were collected from five cities and seven provinces across the six regions in Vietnam. Of these, 900 samples were collected from wet markets and 100 from supermarkets. All samples were analyzed for the presence of Salmonella according to a method recommended by the U. S. Department of Agriculture, Food Safety and Inspection Service. The overall Salmonella prevalence was 45.9%. There was no significant difference (P > 0.05) in Salmonella prevalence by (i) location (Ha Noi city, 51.1%; Hai Phong city, 45.6%; Da Nang and Can Tho cities, 45.5%; Bac Ninh province and Ho Chi Minh city, 44.7%; Dong Nai province, 44.6%; Ha Tinh province, 44.4%; Phu Tho province, 43.8%; Lao Cai province, 43.5%; Kien Giang province, 41.9%; and Lam Dong province, 40.9%), (ii) market type (wet market, 46.2%; supermarket samples, 43.0%), and (iii) storage temperature at retail (ambient storage, 46.4%; chilled storage, 45.1%). Hence, Salmonella presence on poultry meat in Vietnam was not associated with a specific city or province, market type, or storage temperature at retail. Strategies to reduce Salmonella levels on raw poultry in Vietnam should be undertaken to improve the safety of poultry products and reduce the incidence of human salmonellosis from poultry consumption. PMID:23043836

Ta, Yen T; Nguyen, Trung Thanh; To, Phuong Bich; Pham, Da Xuan; Le, Hao Thi Hong; Alali, Walid Q; Walls, Isabel; Lo Fo Wong, Danilo M A; Doyle, Michael P

2012-10-01

310

Salmonella surveillance and control for finisher pigs and pork in Denmark — A case study  

DEFF Research Database (Denmark)

Salmonella can either be controlled pre-harvest, post-harvest or by a combination of both approaches. This paper describes the lessons learned in Danish Salmonella surveillance and control programme for finisher pigs and pork. Initially, main focus was on pre-harvest initiatives and correct identification of herds with respect to the risk for Salmonella that they represented. However, an analysis of risk-mitigating actions applied along the chain from stable to table showed that it would be more cost-effective to deal with Salmonella on the abattoirs than in the herds. This knowledge moved focus from pre- to post-harvest without giving up on pre-harvest surveillance. First of all, this meant increased attention on slaughter hygiene and individual interventions in the abattoirs. In brief, we learned that for a programme to be successful it must be based on standardised methods for sampling and testing to be able to evaluate and compare performance of the programme. More specifically, meat-juice samples taken from finisher pigs at the time of slaughter are an effective way of identifying high-risk herds for Salmonella. In addition, a penalty system might act as an incentive for farmers to deal with Salmonella in their herd. Additionally, common targets for all abattoirs allowing for unique control solutions should be adapted. Finally, decontamination techniques like hot water decontamination are a feasible way of dealing with high-risk pigs (Level-3 pigs). The current prevalence in Danish pork is around 1.2%, and a target is set to

Alban, L.; Baptista, F.M.

2012-01-01

311

Salmonella osteomyelitis of the femoral diaphysis in a healthy individual.  

Science.gov (United States)

In reviewing the literature, we found few cases of Salmonella osteomyelitis of the femoral diaphysis in a healthy patient. Most are typically associated with sickle cell anemia or immunosuppressed patients. We report on the successful treatment of Salmonella osteomyelitis in the mid-diaphyseal region of the femur caused by Salmonella species in a healthy individual. PMID:25303451

Kim, Byoung-Kook; Dan, Jinmyoung; Lee, Yun-Seok; Kim, Seung-Hee; Cha, Yoon-Sik

2014-10-01

312

Diffuse abdominal gallium-67 citrate uptake in salmonella infections  

International Nuclear Information System (INIS)

Two pediatric patients with salmonella infections (one with typhoid fever and the second with salmonella C2 gastroenteritis), had a diffuse abdominal uptake of Ga-67 citrate. The possible explanation for this finding is discussed. Salmonella infection should be included as a cause in the differential diagnosis of diffuse accumulation of Ga-67 citrate

313

Antibiotic Resistant Salmonella and Vibrio Associated with Farmed Litopenaeus vannamei  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Salmonella and Vibrio species were isolated and identified from Litopenaeus vannamei cultured in shrimp farms. Shrimp samples showed occurrence of 3.3% of Salmonella and 48.3% of Vibrio. The isolates were also screened for antibiotic resistance to oxolinic acid, sulphonamides, tetracycline, sulfamethoxazole/trimethoprim, norfloxacin, ampicillin, doxycycline hydrochloride, erythromycin, chloramphenicol, and nitrofurantoin. Salmonella enterica serovar Corvallis isolated from shrimp showed indiv...

Banerjee, Sanjoy; Ooi, Mei Chen; Shariff, Mohamed; Khatoon, Helena

2012-01-01

314

Inhibitory Effects of Chick Lactobacilli on Enteropathogenic Salmonella  

Directory of Open Access Journals (Sweden)

Full Text Available The major source of human salmonellosis are farm animals, which may frequently be intestinal carriers of the organism. Lactobacilli isolated from the intestinal tract with efficiency adhesion were selected and studied in assay competitions with different Salmonella serotypes. The growth and lectin production of lactobacilli remained unchanged in several mixed and single culture studies. However, in mixed cultures, the inhibition (bacteriostatic of viable bacteria of salmonella strains was observed. The adhesion ratio showed significant values for L. animalis, L. fermentum, S. Gallinarum and S. Pullorum. These results indicate the remarkable importance of a specific host interaction in the colonization process by microorganisms. L. animalis was effective in reducing the attachment of S. Gallinarum, S. Pullorum and S. Enteritidis to host-specific epithelial cells, while L. fermentum was able to reduce the attachment of S. Pullorum and S. Gallinarum. Therefore, we suggest that chicken lactobacilli included in this work may be considered potential probiotic microorganisms, and used in the preparation of a probiotic food.

D. Draksler

2006-01-01

315

Salmonella enterica as a vaccine carrier.  

Science.gov (United States)

Salmonella enterica is an invasive, facultative intracellular gastrointestinal pathogen causing human diseases such as gastroenteritis and typhoid fever. Virulence-attenuated strains of this pathogen have interesting capacities for the generation of live vaccines. Attenuated live typhoidal and nontyphoidal Salmonella strains can be used for vaccination against Salmonella infections and to target tumor tissue. Such strains may also serve as live carriers for the development of vaccination strategies against other bacterial, viral or parasitic pathogens. Various strategies have been developed to deploy regulatory circuits and protein secretion systems for efficient expression and delivery of foreign antigens by Salmonella carrier strains. One prominent example is the use of type III secretion systems to translocate recombinant antigens into antigen presenting cells. In this review, we will describe the recent developments in strategies that utilize live attenuated Salmonella as vaccine carriers for prophylactic vaccination against infectious diseases and therapeutic vaccination against tumors. Considerations for generating safe, attenuated carrier strains, designing stable expression systems and the use of adjuvants for live carrier strategies are discussed. PMID:22191450

Hegazy, Wael Abdel Halim; Hensel, Michael

2012-01-01

316

Sensitivity pattern of Salmonella serotypes in Northern India  

Directory of Open Access Journals (Sweden)

Full Text Available BACKGROUND: Enteric fever continues to be a major public health problem, especially in the developing countries of the tropics. We determined the incidence of Salmonella bloodstream infections and their antimicrobial resistance patterns from May to August in the years 1997-2001 in Haryana, a large state of India. The minimum inhibitory concentration (MIC was also determined for 60 isolates of S. typhi to various commonly used antimicrobial agents. MATERIAL AND METHODS: Blood cultures of 6,956 patients (PUO/septicemia were processed by standard procedures and the Salmonella spp. isolates were identified with specific antisera and with standard biochemical tests. Antimicrobial susceptibilities were determined by Stokes disc diffusion method. The MIC of 60 randomly isolated strains of S. typhi was determined by agar dilution method using Mueller Hinton Agar medium. RESULTS: Isolation rates of Salmonella spp. increased in 2000 and 2001. Multidrug resistance (MDR in S. typhi had increased while in S. paratyphi it had decreased markedly. Ninety per cent chloramphenicol sensitivity was seen in S. typhi by MIC method. There was a decrease in the susceptibility to ciprofloxacin of S. typhi with MIC showing an upward trend. All S. typhi tested were sensitive to third generation cephalosporins and aminoglycosides with MIC well below the breakpoint. DISCUSSION: Our study indicates that MDR in S. typhi is on the rise in our area. There is also re-emergence of chloramphenicol sensitivity. Rising MIC values of ciprofloxacin may lead to prolonged treatment, delayed recovery or pose treatment failure. Thus, sensitivity pattern of causative organism must be sought before instituting appropriate therapy to prevent further emergence of drug resistance.

Gautam Vikas

2002-01-01

317

Sensitivity pattern of Salmonella serotypes in Northern India  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english BACKGROUND: Enteric fever continues to be a major public health problem, especially in the developing countries of the tropics. We determined the incidence of Salmonella bloodstream infections and their antimicrobial resistance patterns from May to August in the years 1997-2001 in Haryana, a large s [...] tate of India. The minimum inhibitory concentration (MIC) was also determined for 60 isolates of S. typhi to various commonly used antimicrobial agents. MATERIAL AND METHODS: Blood cultures of 6,956 patients (PUO/septicemia) were processed by standard procedures and the Salmonella spp. isolates were identified with specific antisera and with standard biochemical tests. Antimicrobial susceptibilities were determined by Stokes disc diffusion method. The MIC of 60 randomly isolated strains of S. typhi was determined by agar dilution method using Mueller Hinton Agar medium. RESULTS: Isolation rates of Salmonella spp. increased in 2000 and 2001. Multidrug resistance (MDR) in S. typhi had increased while in S. paratyphi it had decreased markedly. Ninety per cent chloramphenicol sensitivity was seen in S. typhi by MIC method. There was a decrease in the susceptibility to ciprofloxacin of S. typhi with MIC showing an upward trend. All S. typhi tested were sensitive to third generation cephalosporins and aminoglycosides with MIC well below the breakpoint. DISCUSSION: Our study indicates that MDR in S. typhi is on the rise in our area. There is also re-emergence of chloramphenicol sensitivity. Rising MIC values of ciprofloxacin may lead to prolonged treatment, delayed recovery or pose treatment failure. Thus, sensitivity pattern of causative organism must be sought before instituting appropriate therapy to prevent further emergence of drug resistance.

Vikas, Gautam; Naveen Kumar, Gupta; Uma, Chaudhary; D. R., Arora.

318

Report of neonatal meningitis due to Salmonella enterica serotype Agona and review of breast milk-associated neonatal Salmonella infections.  

Science.gov (United States)

We present the first documented case of Salmonella enterica serotype Agona meningitis in a 6-day-old baby. S. enterica serotype Agona was isolated concurrently from infant cerebrospinal fluid and parental fecal samples, and Salmonella was isolated from breast milk. The role of breast milk in transmission of Salmonella enterica is discussed. PMID:19605582

Cooke, Fiona J; Ginwalla, Sara; Hampton, Michael D; Wain, John; Ross-Russell, Robert; Lever, Andrew; Farrington, Mark

2009-09-01

319

Report of Neonatal Meningitis Due to Salmonella enterica Serotype Agona and Review of Breast Milk-Associated Neonatal Salmonella Infections?  

Science.gov (United States)

We present the first documented case of Salmonella enterica serotype Agona meningitis in a 6-day-old baby. S. enterica serotype Agona was isolated concurrently from infant cerebrospinal fluid and parental fecal samples, and Salmonella was isolated from breast milk. The role of breast milk in transmission of Salmonella enterica is discussed. PMID:19605582

Cooke, Fiona J.; Ginwalla, Sara; Hampton, Michael D.; Wain, John; Ross-Russell, Robert; Lever, Andrew; Farrington, Mark

2009-01-01

320

Report of Neonatal Meningitis Due to Salmonella enterica Serotype Agona and Review of Breast Milk-Associated Neonatal Salmonella Infections?  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We present the first documented case of Salmonella enterica serotype Agona meningitis in a 6-day-old baby. S. enterica serotype Agona was isolated concurrently from infant cerebrospinal fluid and parental fecal samples, and Salmonella was isolated from breast milk. The role of breast milk in transmission of Salmonella enterica is discussed.

Cooke, Fiona J.; Ginwalla, Sara; Hampton, Michael D.; Wain, John; Ross-russell, Robert; Lever, Andrew; Farrington, Mark

2009-01-01

 
 
 
 
321

Hemagglutinating properties of Salmonella enterica serovar Enteritidis isolated from different sources / Propriedades hemaglutinantes de Salmonella enterica sorotipo Enteritidis isoladas de diferentes fontes  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese Foram estudadas 25 amostras de Salmonella enterica sorotipo Enteritidis isoladas de diferentes fontes, em testes de hemaglutinação. Amostras bacterianas cultivadas em diferentes meios de cultura causavam hemaglutinação na presença de hemácias humanas, entretanto, não foi observada reação com hemácia [...] s de outras espécies. A expressão da atividade hemaglutinante foi melhor em ágar CFA a 37ºC. A hemaglutinação foi inibida por D-manose, D-manitol, melibiose, D-rafinose, L-ramnose e sacarose. A análise ultraestrutural não revelou a presença de estruturas filamentosas na superfície bacteriana, sugerindo que a hemaglutinina de Salmonella Enteritidis seja de natureza não fimbrial. Os dados sugerem que Salmonella Enteritidis produz uma hemaglutinina não fimbrial manose-sensível, específica para hemácias humanas, que pode ser extraída na forma solúvel. Abstract in english Twenty-five strains of Salmonella enterica serovar Enteritidis isolated from different sources were examined for hemagglutinating activity. Bacteria cultured in different media induced hemagglutination of human erythrocytes, but no reaction was observed with erythrocytes from other animal species. T [...] he hemagglutinating expression activity was better for cultures on CFA agar at 37ºC than other conditions examined. The hemagglutination was inhibited by D-mannose, D-mannitol, melibiose, D-raffinose, L-rhamnose and sucrose. The absence of cell-surface appendages in electron microscope examinations suggested a nonfimbrial hemagglutinin. The data suggest that Salmonella Enteritidis produces nonfimbrial mannose-sensitive hemagglutinin, specific for human erythrocytes, which could be extracted in soluble form.

Jane M.G., Mikcha; Antonio J. Piantino, Ferreira; Claudete S. Astolfi, Ferreira; Tomomasa, Yano.

322

The type VI secretion system encoded in Salmonella pathogenicity island 19 is required for Salmonella enterica serotype Gallinarum survival within infected macrophages.  

Science.gov (United States)

Salmonella enterica serotype Gallinarum is the causative agent of fowl typhoid, a disease characterized by high morbidity and mortality that causes major economic losses in poultry production. We have reported that S. Gallinarum harbors a type VI secretion system (T6SS) encoded in Salmonella pathogenicity island 19 (SPI-19) that is required for efficient colonization of chicks. In the present study, we aimed to characterize the SPI-19 T6SS functionality and to investigate the mechanisms behind the phenotypes previously observed in vivo. Expression analyses revealed that SPI-19 T6SS core components are expressed and produced under in vitro bacterial growth conditions. However, secretion of the structural/secreted components Hcp1, Hcp2, and VgrG to the culture medium could not be determined, suggesting that additional signals are required for T6SS-dependent secretion of these proteins. In vitro bacterial competition assays failed to demonstrate a role for SPI-19 T6SS in interbacterial killing. In contrast, cell culture experiments with murine and avian macrophages (RAW264.7 and HD11, respectively) revealed production of a green fluorescent protein-tagged version of VgrG soon after Salmonella uptake. Furthermore, infection of RAW264.7 and HD11 macrophages with deletion mutants of SPI-19 or strains with genes encoding specific T6SS core components (clpV and vgrG) revealed that SPI-19 T6SS contributes to S. Gallinarum survival within macrophages at 20 h postuptake. SPI-19 T6SS function was not linked to Salmonella-induced cytotoxicity or cell death of infected macrophages, as has been described for other T6SS. Our data indicate that SPI-19 T6SS corresponds to a novel tool used by Salmonella to survive within host cells. PMID:23357385

Blondel, Carlos J; Jiménez, Juan C; Leiva, Lorenzo E; Alvarez, Sergio A; Pinto, Bernardo I; Contreras, Francisca; Pezoa, David; Santiviago, Carlos A; Contreras, Inés

2013-04-01

323

Hemagglutinating properties of Salmonella enterica serovar Enteritidis isolated from different sources / Propriedades hemaglutinantes de Salmonella enterica sorotipo Enteritidis isoladas de diferentes fontes  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese Foram estudadas 25 amostras de Salmonella enterica sorotipo Enteritidis isoladas de diferentes fontes, em testes de hemaglutinação. Amostras bacterianas cultivadas em diferentes meios de cultura causavam hemaglutinação na presença de hemácias humanas, entretanto, não foi observada reação com hemácia [...] s de outras espécies. A expressão da atividade hemaglutinante foi melhor em ágar CFA a 37ºC. A hemaglutinação foi inibida por D-manose, D-manitol, melibiose, D-rafinose, L-ramnose e sacarose. A análise ultraestrutural não revelou a presença de estruturas filamentosas na superfície bacteriana, sugerindo que a hemaglutinina de Salmonella Enteritidis seja de natureza não fimbrial. Os dados sugerem que Salmonella Enteritidis produz uma hemaglutinina não fimbrial manose-sensível, específica para hemácias humanas, que pode ser extraída na forma solúvel. Abstract in english Twenty-five strains of Salmonella enterica serovar Enteritidis isolated from different sources were examined for hemagglutinating activity. Bacteria cultured in different media induced hemagglutination of human erythrocytes, but no reaction was observed with erythrocytes from other animal species. T [...] he hemagglutinating expression activity was better for cultures on CFA agar at 37ºC than other conditions examined. The hemagglutination was inhibited by D-mannose, D-mannitol, melibiose, D-raffinose, L-rhamnose and sucrose. The absence of cell-surface appendages in electron microscope examinations suggested a nonfimbrial hemagglutinin. The data suggest that Salmonella Enteritidis produces nonfimbrial mannose-sensitive hemagglutinin, specific for human erythrocytes, which could be extracted in soluble form.

Jane M.G., Mikcha; Antonio J. Piantino, Ferreira; Claudete S. Astolfi, Ferreira; Tomomasa, Yano.

2004-06-01

324

Development of a novel cross-streaking method for isolation, confirmation, and enumeration of Salmonella from irrigation ponds.  

Science.gov (United States)

The 2013 Produce Safety Rules in Food Safety Modernization Act (FSMA) require regular testing for generic Escherichia coli in agricultural water intended for pre-harvest contact with the edible portion of fresh produce. However, the use of fecal contamination indicators frequently does not correctly reflect distribution of foodborne pathogens such as Salmonella enterica, and ensuring food safety may require direct detection and enumeration of pathogens in agricultural settings. Herein we report the evaluation of different cost-effective methods for quantification, isolation, and confirmation of Salmonella in irrigation pond water and sediment samples. A most probably number (MPN) dual enrichment culture method was used in combination with differential and selective agars, XLT4 and CHROMagar™ Salmonella plus (CSP). The necessity for PCR confirmation was evaluated, and methods were compared by cost and performance measures (i.e., sensitivity, specificity, positive predictive value, and negative predictive value). Statistical analyses showed that using XLT4 as the initial selective agar to isolate Salmonella colonies improved recovery compared to CSP agar; however, PCR confirmation was required to avoid false positive results on either agar. Therefore, a novel cross-streaking method utilizing CHROMagar™ agar for individual colony confirmation of Salmonella presence/absence on XLT4 was developed. This method classifies the colony as positive if typical Salmonella appearance is observed on both agars. Statistical analysis showed that this method was as effective as PCR for species confirmation of pure individual strains isolated from enrichment cultures (sensitivity=0.99, specificity=1.00, relative to PCR). This method offers a cost-effective alternative to PCR that would increase the capacity and sensitivity of Salmonella evaluation. PMID:24732066

Luo, Zhiyao; Gu, Ganyu; Giurcanu, Mihai C; Adams, Paige; Vellidis, George; van Bruggen, Ariena H C; Wright, Anita C

2014-06-01

325

Multilocus sequence typing supports the hypothesis that cow- and human-associated Salmonella isolates represent distinct and overlapping populations.  

Science.gov (United States)

A collection of 179 human and 156 bovine clinical Salmonella isolates obtained from across New York state over the course of 1 year was characterized using serotyping and a multilocus sequence typing (MLST) scheme based on the sequencing of three genes (fimA, manB, and mdh). The 335 isolates were differentiated into 52 serotypes and 72 sequence types (STs). Analyses of bovine isolates collected on different farms over time indicated that specific subtypes can persist over time on a given farm; in particular, a number of farms showed evidence for the persistence of a specific Salmonella enterica serotype Newport sequence type. Serotypes and STs were not randomly distributed among human and bovine isolates, and selected serotypes and STs were associated exclusively with either human or bovine sources. A number of common STs were geographically widespread. For example, ST6, which includes isolates representing serotype Typhimurium as well as the emerging serotype 4,5,12:i:-, was found among human and bovine isolates in a number of counties in New York state. Phylogenetic analyses supported the possibility that serotype 4,5,12:i:- is closely related to Salmonella serotype Typhimurium. Salmonella serotype Newport was found to represent two distinct evolutionary lineages that differ in their frequencies among human and bovine isolates. A number of Salmonella isolates carried two copies of manB (33 isolates) or showed small deletion events in fimA (nine isolates); these duplication and deletion events may provide mechanisms for the rapid diversification of Salmonella surface molecules. We conclude that the combined use of an economical three-gene MLST scheme and serotyping can provide considerable new insights into the evolution and transmission of Salmonella. PMID:17028236

Alcaine, S D; Soyer, Y; Warnick, L D; Su, W-L; Sukhnanand, S; Richards, J; Fortes, E D; McDonough, P; Root, T P; Dumas, N B; Gröhn, Y; Wiedmann, M

2006-12-01

326

Comparison of 3 culture methods and PCR assays for Salmonella gallinarum and Salmonella pullorum detection in poultry feed.  

Science.gov (United States)

To detect Salmonella gallinarum or Salmonella pullorum in artificially contaminated poultry feed, 9 culture combinations were compared, including 3 preenrichment/enrichment methods (tryptic soy broth plus ferrous sulfate/tetrathionate Hajna, tryptic soy broth plus ferrous sulfate/selenite cystine broth, and Salmosyst) in combination with 3 selective agars (xylose lysine desoxicholate agar added with tergitol 4, EF-18, and Önöz), respectively. Additionally, a single PCR technique was applied combined with 2 different preenrichment media (tryptic soy broth plus ferrous sulfate and Salmosyst). The specificity and positive predictive value were 1 for all methods. There were some differences among Salmonella strains for sensitivity and accuracy in the culture and Salmosyst-PCR methods. The sensitivity and accuracy values were less than 0.60 and 0.64, respectively, whereas the negative predictive values were between 0.12 and 0.23. Two PCR methods did not show any difference in the parameters of performance evaluated. Kappa coefficients showed good agreement between both methods. None of the culture combinations was able to detect S. gallinarum or S. pullorum when the inoculum was less than 3 × 10² cfu/25 g, except the Salmosyst broth method, which could recover S. gallinarum from 3 × 10¹ cfu/25 g onward. Overall, there were differences in the detection limits among the strains and methods used. In general, the 3 selective plating media did not show any significant difference in the parameters of performance studied for each strain. On the other hand, the agreements were slight to fair when culture methods were compared among them and with both PCR methods. The differences in the detection levels that were obtained using these methods and the difficulty in detecting S. gallinarum or S. pullorum in feed represent a potential problem when a poultry feed sample is considered to be negative. It is highly recommended to use at least 2 methods to increase the chances of detecting S. gallinarum or S. pullorum in poultry feed. PMID:23687146

Soria, M Cecilia; Soria, Mario A; Bueno, Dante J; Terzolo, Horacio R

2013-06-01

327

Inactivation of Salmonellae in Frozen Catfish by Gamma Irradiation  

International Nuclear Information System (INIS)

The effect of gamma irradiation on salmonellae viability in frozen catfish was investigated using fresh cut of catfish artificially contaminated with stationary phase cells of salmonellae, frozen at-18 ?C and irradiated with does ranging from 0.0 to 2.4 kGy. The D10 values for ten serovars of salmonellae ranged from 0.47 to 0.77 kGy. Salmonella Enteritidis was the most resistant serovars found in frozen catfish. Dosage at 2.5 kGy would be sufficient to kill 103.2 Salmonella Enteritidis that may occasionally present in frozen catfish

328

Septic arthritis of the ankle due to Salmonella enteritidis.  

LENUS (Irish Health Repository)

Salmonella septic arthritis in healthy, immunocompetent patients is extremely rare. We present the case of a 70-year-old man who presented with a one-day history of painful swelling of his ankle from which was aspirated pus which subsequently grew Salmonella enteritidis. There was no history of trauma or symptoms consistent with Salmonella enterocolitis. Our patient recovered fully after two weeks on intravenous ceftriaxone and six weeks on oral ciprofloxacin. Salmonella is a notifiable disease in the European Union and the United States of America, and is associated with outbreaks as a result of food contamination. The nature of Salmonella arthritis and its appropriate management are outlined.

Dineen, Patrick F

2011-06-01

329

Salmonellae in foods and animal feeding stuffs  

International Nuclear Information System (INIS)

After the problem of Salmonella infections in foods and feeding stuffs is emphasized, an account is given of the current ways of manufacturing bone meal, meat meal, blood meal, fish meal, fish flour, egg products and coconut. The effectiveness in eliminating salmonellae and the chance and possible sources of recontamination are described for each production method. Besides heat treatment, fumigation by ethylene oxide and irradiation with gamma rays are considered. The bacteriological tests required to establish the effectiveness of treatment are also discussed, as well as the effect of the treatment on the nutritive value of the product. (author). 50 refs, 4 tabs

330

Salmonella in Brazilian and imported pet reptiles  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The presence of salmonellae in fecal samples or cloacal swabs of 97 pet reptiles (15 snakes, 24 lizards and 58 chelonians) was investigated. Thirty seven animals had national origin and 60 were imported. Salmonella spp was detected in 39.1% of the reptiles, being 62.5% in lizards, 53.3% in snakes and 25.8% in chelonians. Strains belonged to subspecies I (44.7%), II (10.5%), IIIa (5.2%), IIIb (21.0%) and IV (18.5%) of the enterica species, with predominance (55.3%) of subspecies usually found ...

Sá Isabel Valéria Abalem de; Solari Claude André

2001-01-01

331

Rapid radiometric method for detection of Salmonella in foods  

Energy Technology Data Exchange (ETDEWEB)

A radiometric method for the detection of Salmonella in foods has been developed which is based on Salmonella poly H agglutinating serum preventing Salmonella from producing 14CO2 from (14C) dulcitol. The method will detect the presence or absence of Salmonella in a product within 30 h compared to 4 to 5 days by routine culture methods. The method has been evaluated against a routine culture method using 58 samples of food. The overall agreement was 91%. Five samples negative for Salmonella by the routine method were positive by the radiometric method. These may have been false positives. However, the routine method may have failed to detect Salmonella due to the presence of large numbers of lactose-fermenting bacteria which hindered isolation of Salmonella colonies on the selective agar plates.

Stewart, B.J.; Eyles, M.J.; Murrell, W.G.

1980-08-01

332

Distribution of Salmonella serovars and antimicrobial susceptibility of Salmonella Enteritidis from poultry in Zimbabwe.  

Science.gov (United States)

A study was carried out to determine the distribution and antimicrobial susceptibility profiles of Salmonella serovars from chickens from large-scale commercial (LSC), small-scale commercial (SSC), and rural free-range (RFR) farms of Zimbabwe. Pooled cloacal swabs were collected for culture and isolation of Salmonella spp. A chi-square test was used to assess distribution differences of salmonellas among the farming sectors. Approximately 10% (283/2833) of the swabs were positive for Salmonella enterica, with only subspecies enterica (98.6%) and arizonae (1.4%) being detected. The prevalence of S. enterica varied significantly (Pfurazolidone, neomycin and trimethoprim-sulfamethoxazole. There were 12.1% multi-drug-resistant S. Enteritidis isolates, and the resistance to ampicillin/kanamycin was predominant. The identification of multi-drug-resistant S. Enteritidis is of public health concern. Thus, stringent control of S. Enteritidis will reduce the public health risk of human salmonellosis. PMID:22515540

Makaya, P V; Matope, G; Pfukenyi, D M

2012-01-01

333

Mionecrosis masiva por salmonella enteritidis en paciente inmunodeprimido Massive salmonella enteritidis myonecrosis in inmunodeficient pacient  

Directory of Open Access Journals (Sweden)

Full Text Available Salmonella enteritidis es un microorganismo que raramente afecta al componente muscular. Recogemos el caso de un paciente varón con Síndrome de Inmunodeficiencia Adquirida (SIDA, que tras un episodio de gastroenteritis aguda debida a Salmonella enteritidis presenta, varios meses después, una necrosis muscular masiva en los miembros inferiores, que le llevó al shock séptico y posteriormente a la muerte.Salmonella enteritidis is a microorganism which rarely affects muscles. We report a case of a male patient with AIDS, who suffered an episode of acute gastroenteritis due to Salmonella enteritidis, months ago. After this incident, the patient presented massive muscular necrosis in the lower limbs which led to septic shock and subsequent death.

P. Baeta

2007-06-01

334

Empiema causado por Salmonella typhimurium / Pleural empyema caused by Salmonella typhimurium: A case report  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: Spanish Abstract in spanish El género Salmonella se caracteriza por causar infecciones en el tracto gastrointestinal, debido a la ingesta de alimentos o agua contaminada. También puede causar, con menor frecuencia, infecciones localizadas en diferentes órganos; esto se asocia con inmunodepresión. En este caso se describe un pa [...] ciente con infección pleuropulmonar por Salmonella typhimurium, que no reportó antecedentes de diarrea previa. Evolucionó favorablemente con tratamiento adecuado. Abstract in english Salmonella species are commonly associated with acute gastroenteritis due to ingestion of contaminated food or water. Extraintestinal infections are less frequent, and most of them occur in immunocompromised patients. We report a case of pleural empyema caused by Salmonella typhimurium, without prev [...] ious diarrhea or fever. The patient evolved favorably after receiving adequate treatment.

MARÍA DEL MAR, TACCHINI A; ANA, CARAFFINI F; MARÍA SOLEDAD, MONTAMAT C; NATALIA, SPITALE A; YANINA, BOSIO D; ANGEL, MINGUEZ G.

335

Empiema causado por Salmonella typhimurium Pleural empyema caused by Salmonella typhimurium: A case report  

Directory of Open Access Journals (Sweden)

Full Text Available El género Salmonella se caracteriza por causar infecciones en el tracto gastrointestinal, debido a la ingesta de alimentos o agua contaminada. También puede causar, con menor frecuencia, infecciones localizadas en diferentes órganos; esto se asocia con inmunodepresión. En este caso se describe un paciente con infección pleuropulmonar por Salmonella typhimurium, que no reportó antecedentes de diarrea previa. Evolucionó favorablemente con tratamiento adecuado.Salmonella species are commonly associated with acute gastroenteritis due to ingestion of contaminated food or water. Extraintestinal infections are less frequent, and most of them occur in immunocompromised patients. We report a case of pleural empyema caused by Salmonella typhimurium, without previous diarrhea or fever. The patient evolved favorably after receiving adequate treatment.

MARÍA DEL MAR TACCHINI A

2010-06-01

336

Salmonella Typhi shdA: pseudogene or allelic variant?  

Science.gov (United States)

ShdA from Salmonella Typhimurium (ShdASTm) is a large outer membrane protein that specifically recognizes and binds to fibronectin. ShdASTm is involved in the colonization of the cecum and the Peyer's patches of terminal ileum in mice. On the other hand, shdA gene from Salmonella Typhi (shdASTy) has been considered a pseudogene (i.e. a nonfunctional sequence of genomic DNA) due to the presence of deletions and mutations that gave rise to premature stop codons. In this work we show that, despite the deletions and mutations, shdASTy is fully functional. S. Typhi ?shdA mutants presented an impaired adherence and invasion of HEp-2 pre-treated with TGF-?1, an inducer of fibronectin production. Moreover, shdA from S. Typhi and S. Typhimurium seem to be equivalent since shdASTm restored the adherence and invasion of S. Typhi ?shdA mutant to wild type levels. In addition, anti-FLAG mAbs interfered with the adherence and invasion of the S. Typhi shdA-3xFLAG strain. Finally, shdASTy encodes a detectable protein when heterologously expressed in Escherichia coli DH5?. The data presented here show that shdASTy is not a pseudogene, but a different functional allele compared with shdASTm. PMID:24859062

Urrutia, I M; Fuentes, J A; Valenzuela, L M; Ortega, A P; Hidalgo, A A; Mora, G C

2014-08-01

337

Effects of lipoprotein biogenesis mutations on flagellar assembly in Salmonella.  

Science.gov (United States)

Flagellar assembly requires the expression of a large number of flagellum-specific genes. However, mutations in a number of other genes in Salmonella and Escherichia coli have been shown to have pleiotropic effects that affect flagellar assembly. FlgH (the L-ring subunit of the flagellar basal body) is a lipoprotein whose modification is important for L-ring assembly. We therefore tested whether the lack of motility of Salmonella mutants defective in lipoprotein biogenesis is a result of inability to modify FlgH. Our results show that temperature-sensitive apolipoprotein N-acyltransferase [lnt(Ts)] mutants are nonflagellate at 42 degrees C. However, the flagellar assembly defect occurs at a much earlier step in the pathway than L-ring assembly. These mutants failed to assemble even an MS ring, presumably because of the observed decrease in transcription of fliF. In contrast, temperature-sensitive diacylglycerol transferase [lgt(Ts)] mutants were motile at 42 degrees C, provided the strains carried an lpp (Braun lipoprotein) mutation to permit growth. We have isolated second-site mutants from an lgt(Ts) lpp(+) strain that grow but are nonflagellate at 42 degrees C. Thus, lipoprotein biogenesis is a factor that is important for flagellar assembly. PMID:11790747

Dailey, Frank E; Macnab, Robert M

2002-02-01

338

Evaluation of the respiratory route as a viable portal of entry for Salmonella in poultry via intratracheal challenge of Salmonella Enteritidis and Salmonella Typhimurium.  

Science.gov (United States)

Experimental and epidemiological evidence suggests that primary infection of Salmonella is by the oral-fecal route for poultry. However, the airborne transmission of Salmonella and similar enteric zoonotic pathogens has been historically neglected. Increasing evidence of Salmonella bioaerosol generation in production facilities and studies suggesting the vulnerabilities of the avian respiratory architecture together have indicated the possibility of the respiratory system being a potential portal of entry for Salmonella in poultry. Presently, we evaluated this hypothesis through intratracheal (IT) administration of Salmonella Enteritidis and Salmonella Typhimurium, as separate challenges, in a total of 4 independent trials, followed by enumeration of cfu recovery in ceca-cecal tonsils and recovery incidence in liver and spleen. In all trials, both Salmonella Enteritidis and Salmonella Typhimurium, challenged IT colonized cecae to a similar or greater extent than oral administration at identical challenge levels. In most trials, chickens cultured for cfu enumeration from IT-challenged chicks at same dose as orally challenged, resulted in an increase of 1.5 log higher Salmonella Enteritidis from ceca-cecal tonsils and a much lower dose IT of Salmonella Enteritidis could colonize ceca to the same extent than a higher oral challenge. This trend of increased cecal colonization due to IT challenge was observed with all trails involving week-old birds (experiment 2 and 3), which are widely considered to be more difficult to infect via the oral route. Liver-spleen incidence data showed 33% of liver and spleen samples to be positive for Salmonella Enteritidis administered IT (10(6) cfu/chick), compared with 0% when administered orally (experiment 2, trial 1). Collectively, these data suggest that the respiratory tract may be a largely overlooked portal of entry for Salmonella infections in chickens. PMID:24570455

Kallapura, G; Morgan, M J; Pumford, N R; Bielke, L R; Wolfenden, A D; Faulkner, O B; Latorre, J D; Menconi, A; Hernandez-Velasco, X; Kuttappan, V A; Hargis, B M; Tellez, G

2014-02-01

339

Paradigm Diagnostics Salmonella Indicator Broth (PDX-SIB) for detection of Salmonella on selected environmental surfaces.  

Science.gov (United States)

The Paradigm Diagnostics Salmonella Indicator Broth (PDX-SIB) is intended as a single-step selective enrichment indicator broth to be used as a simple screening test for the presence of Salmonella spp. in environmental samples. This method permits the end user to avoid multistep sample processing to identify presumptively positive samples, as exemplified by standard U.S. reference methods. PDX-SIB permits the outgrowth of Salmonella while inhibiting the growth of competitive Gram-negative and -positive microflora. Growth of Salmonella-positive cultures results in a visual color change of the medium from purple to yellow when the sample is grown at 37 +/- 1 degree C. Performance of PDX-SIB has been evaluated in five different categories: inclusivity-exclusivity, methods comparison, ruggedness, lot-to-lot variability, and shelf stability. The inclusivity panel included 100 different Salmonella serovars, 98 of which were SIB-positive during the 30 to 48 h incubation period. The exclusivity panel included 33 different non-Salmonella microorganisms, 31 of which were SIB-negative during the incubation period. Methods comparison studies included four different surfaces: S. Newport on plastic, S. Anatum on sealed concrete, S. Abaetetuba on ceramic tile, and S. Typhimurium in the presence of 1 log excess of Citrobacter freundii. Results of the methods comparison studies demonstrated no statistical difference between the SIB method and the U.S. Food and Drug Administration-Bacteriological Analytical Manual reference method, as measured by the Mantel-Haenszel Chi-square test. Ruggedness studies demonstrated little variation in test results when SIB incubation temperatures were varied over a 34-40 degrees C range. Lot-to-lot consistency results suggest no detectable differences in manufactured goods using two reference Salmonella serovars and one non-Salmonella microorganism. PMID:23767367

Olstein, Alan; Griffith, Leena; Feirtag, Joellen; Pearson, Nicole

2013-01-01

340

Variant Salmonella Genomic Island 1 Antibiotic Resistance Gene Cluster in Salmonella enterica Serovar Albany  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Salmonella genomic island 1 (SGI1) contains an antibiotic resistance gene cluster and has been previously identified in multidrug-resistant Salmonella enterica serovars Typhimurium DT104, Agona, and Paratyphi B. We identified a variant SGI1 antibiotic-resistance gene cluster in a multidrug-resistant strain of S. enterica serovar Albany isolated from food fish from Thailand and imported to France. In this strain, the streptomycin resistance aadA2 gene cassette in one of the SGI1 integrons was ...

Doublet, Benoi?t; Lailler, Renaud; Meunier, Danie?le; Brisabois, Anne; Boyd, David; Mulvey, Michael R.; Chaslus-dancla, Elisabeth; Cloeckaert, Axel

2003-01-01

 
 
 
 
341

Variant Salmonella Genomic Island 1-L Antibiotic Resistance Gene Cluster in Salmonella enterica Serovar Newport?  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We report a new Salmonella genomic island 1 variant antibiotic resistance gene cluster called SGI1-L in a Salmonella enterica serovar Newport isolate containing a dfrA15 gene cassette conferring resistance to trimethoprim. The isolate carried another class 1 integron containing the aacC5 and aadA7 gene cassettes conferring resistance to gentamicin and streptomycin/spectinomycin, respectively.

Cloeckaert, Axel; Praud, Karine; Doublet, Benoi?t; Demartin, Marie; Weill, Franc?ois-xavier

2006-01-01

342

Cytotoxic T cell adjuvant effects of three Salmonella enterica flagellins Efeitos adjuvantes para células T citotóxicas de três flagelinas de Salmonella enterica  

Directory of Open Access Journals (Sweden)

Full Text Available Bacterial flagellins are important virulence-associated factors and strong inducers of inflammatory responses in mammalian hosts. Flagellins have also been investigated as potential vaccine adjuvants, either for induction of humoral or cellular immune responses, to different target antigens. In this study we investigated the adjuvant properties of three Salmonella enterica flagellins types (FliCd, FliCi and FljB to an ovalbumin-derived CD8+ T cell-restricted epitope (OVA257264. Although mice immunized with the three tested flagellins elicited antigen-specific activated CD8+ T cells, only animals immunized with FliCi and FliCd flagellins admixed with ovalbumin mounted specific in vivo cytotoxic responses to peptide-pulsed target cells. The present results indicate that Salmonella flagellins are endowed with type-specific adjuvant effects toward murine CD8+ T cells, a feature that may impact their use as adjuvants for prophylatic or therapeutic vaccines.As flagelinas bacterianas são importantes fatores associados à virulência e potentes indutores de resposta inflamatória em mamíferos. Estas moléculas são também investigadas como potencial adjuvante para uso em vacinas na indução de resposta imune humoral e celular para diferentes antígenos alvo. No presente estudo investigamos as propriedades adjuvantes de três tipos de flagelinas de Salmonella enterica (FliCd, FliCi e FljB para um epítopo derivado da ovalbumina específico para células T CD8+. As três flagelinas testadas induziram respostas de células T CD8+ específicas em camundongos imunizados, porém, somente animais imunizados com as flagelinas FliCi e FliCd co-administradas com ovalbumina montaram resposta citotóxica específica in vivo para células-alvo pulsadas com peptídeo OVA. Os resultados apresentados indicam que flagelinas de Salmonella são dotadas de efeitos adjuvantes tipo-específico frente a células T CD8+ in vivo, uma característica que pode gerar impactos no uso dessas proteínas como adjuvantes em vacinas profiláticas ou terapêuticas.

Catarina J.M. Braga

2008-03-01

343

Prediction of Salmonella carcass contamination by a comparative quantitative analysis of E. coli and Salmonella during pig slaughter  

DEFF Research Database (Denmark)

Faecal contamination of carcasses in the slaughterhouse is generally considered to be the source of Salmonella on pork. In this study the hygiene indicator Escherichia coli is used to quantify faecal contamination of carcasses and it is hypothesized that it can be used to predict the quantitative carcass contamination with Salmonella, when the distribution of Salmonella concentrations in faeces is known. Paired pig sample data (faecal samples and carcass swabs) were obtained from five slaughterhouses and analysed for prevalence and concentrations of E. coli and Salmonella. A simple model was developed to describe the faecal contamination of carcasses using the E. coli data. The E. coli results suggested different hygiene performances in different slaughterhouses, and showed that a model assuming that carcasses are predominantly contaminated by their own faeces was not appropriate. Observed Salmonella prevalences were low (on average 1.9% on carcasses) and between slaughterhouses the prevalences ranked differently than the hygiene performance based on the E. coli data suggested. Also, the Salmonella concentrations predicted using E. coli as a faecal indicator were lower than the observed Salmonella concentrations. It is concluded that the faecal carriage of Salmonella together with the faecal contamination of carcasses, as predicted from E. coli data in the animal faeces and hygiene performance of the slaughterhouse, is not sufficient to explain carcass contamination with Salmonella. Our extensive data set showed that other factors than the observed faecal carriage of Salmonella by the individual animals brought to slaughter, play a more important role in the Salmonella carcass contamination of pork.

Nauta, Maarten; Barfod, Kristen

2013-01-01

344

Immunogenic properties of the Salmonella atypical fimbriae in Balb/c mice.  

DEFF Research Database (Denmark)

Components of the Salmonella atypical fimbriae (Saf) were investigated for potential inclusion in a Salmonella vaccine. Recombinant histidine-tagged SafB chaperone complexed with SafD adhesin was expressed in Escherichia coli and purified. Starch microparticles were used, as an adjuvant and recombinant cholera toxin B subunit (rCTB) was included as a mucosal antigen-uptake enhancer. BALB/c mice were immunized orally or subcutaneously with SafB/D- and rCTB-conjugated microparticles and nasally or subcutaneously with SafB/D mixedwith rCTB. The systemic and mucosal immune responses were studied, and an oral challenge with Salmonella enteritidis was performed. All immunized groups except that receiving oral immunization responded with high IgM-IgG titers to SafB/D. Analysis of the subclass ratio (IgG1/IgG2a + IgG2b) indicated a mixed Th1 and Th2 response, with Th1 predominating. The mucosal response, measured as specific IgA/total IgA (from fecal samples), was significantly greater than that in the untreated control group only in the group receiving intranasal immunization (P < 0.05). Spleens were removed 6 days after oral challenge and Salmonella colony-forming units (CFU) were counted. The group immunized subcutaneously with SafB/D- and rCTB-conjugated microparticles had significantly lower CFU counts than the untreated control group (P < 0.05).

Strindelius, Lena; Folkesson, Anders

2004-01-01

345

Evaluation of a serological Salmonella Mix-ELISA for poultry used in a national surveillance programme  

DEFF Research Database (Denmark)

A Mix-ELISA using lipopolysaccharide antigens from Salmonella enterica serotype Enteritidis and Typhimurium? was evaluated using samples collected over time in the Danish salmonella surveillance programme for poultry. Serological samples (n = 42813) taken from broiler-breeder flocks after a year of bacteriological monitoring with negative results were used for calculating the flock and individual test specificities, which were 0.997 and 0.999, respectively. Layer flocks from the table egg sector were used for calculation of positive predictive values. In the survey, flocks were examined for salmonella by Mix-ELISA and by faecal culture, and in case of a positive result in either of these a repeated, serological testing was performed, and 60 animals were organ-cultured. If one of these samplings was positive, the flock was declared salmonella infected. In a period of 3 months, 35 flocks were found to be positive in the routine samples. Of these, 32 were serologically positive, 2 both serologically and faecallypositive and 1 flock only faecally positive. For flocks serologically positive in the surveillance programme, a positive-predictive value of 0.62 for organ culture positivity was found, and while considering serological follow-up samples, the value was 0.95.

Feld, Niels Christian; Ekeroth, Lars

2000-01-01

346

Serological Survey Of Salmonella gallinarum Antibody In Chickens Around Jos, Plateau State, Nigeria  

Directory of Open Access Journals (Sweden)

Full Text Available A serological survey of the prevalence of antibodies to Salmonella gallinarum among chickens under two different management systems around Jos, Plateau State, Nigeria was carried out using the standard plate agglutination test. The objective of this study was to determine serologically the prevalence of antibodies against Salmonella gallinarum among apparently healthy chickens around Jos. A total of 700 serum samples made up of 450 exotic and 250 local breed of chickens were used for this study with 37.9% seropositvity. In the free range system (19.3% of the flocks samp