Sample records for actinobacillus pleuropneumoniae serotypes

  1. Serotyping of Actinobacillus pleuropneumoniae serotype 5 strains using a monoclonal-based polystyrene agglutination test

    Dubreuil, J.D.; Letellier, A.; Stenbæk, Eva; Gottschalk, M.


    A polystyrene agglutination test has been developed for serotyping Actinobacillus pleuropneumoniae serotype 5a and 5b strains. Protein A-coated polystyrene microparticles were sensitized with a murine monoclonal antibody recognizing an epitope on serotype 5 LPS-O chain as shown by SDS-PAGE and...

  2. PCR specific for Actinobacillus pleuropneumoniae serotype 3

    Zhou, L.; Jones, S.C.P.; Angen, Øystein;


    , but the method has liminations, for example, cross-reactions between serotypes 3, 6, and 8. This study describes the development of a serotype 3-specific PCR, based on the capsule locus, which can be used in a multiplex format with the organism's specific gene apxIV. The PCR test was evaluated on 266...

  3. Serological characterization of Actinobacillus pleuropneumoniae biotype 1 strains antigenically related to both serotypes 2 and 7

    Nielsen, R.; Andresen, Lars Ole; Plambeck, Tamara


    Nine Danish Actinobacillus pleuropneumoniae biotype 1 isolates were shown by latex agglutination and indirect haemagglutination to possess capsular polysaccharide epitopes identical to those of serotype 2 strain 1536 (reference strain of serotype 2) and strain 4226 (Danish serotype 2 strain). Imm...

  4. The genetic organization of the capsular polysaccharide biosynthesis region of Actinobacillus pleuropneumoniae serotype 14

    Ito, Hiroya


    The genetic organization of the gene involved in the capsular polysaccharide (CPS) biosynthesis of Actinobacillus pleuropneumoniae serotype 14 has been determined. The DNA region for the CPS biosynthesis of serotype 14 (cps14) comprised 9 open reading frames, designated as cps14AB1B2B3CDEFG genes, encoding Cps14A to Cps14G protein, respectively. Cps14A was similar to CpsA of A. pleuropneumoniae serotypes 1, 4 and 12; the Cps14B1 and Cps14B2 were similar to CpsB of A. pleuropneumoniae serotype...

  5. Experimental vaccination of pigs with an Actinobacillus pleuropneumoniae serotype 5b capsular polysaccharide tetanus toxoid conjugate

    Andresen, Lars Ole; Jacobsen, M.J.; Nielsen, J.P.


    The protective efficacy of an Actinobacillus pleuropneumoniae serotype 5b capsular polysaccharide-tetanus toroid conjugate (Ap5bCP-TT) against homologous challenge of pigs was investigated. Four pigs were non-vaccinated controls (group A), 4 pigs were injected with adjuvant without antigen (group...

  6. Experimental vaccination of pigs with an Actinobacillus pleuropneumoniae serotype 5b capsular polysaccharide tetanus toxoid conjugate

    Andresen, Lars Ole; Jacobsen, M.J.; Nielsen, J.P.


    The protective efficacy of an Actinobacillus pleuropneumoniae serotype 5b capsular polysaccharide-tetanus toroid conjugate (Ap5bCP-TT) against homologous challenge of pigs was investigated. Four pigs were non-vaccinated controls (group A), 4 pigs were injected with adjuvant without antigen (group B...

  7. Isolation of Actinobacillus pleuropneumoniae serotype 2 by immunomagnetic separation

    Angen, Øystein; Heegaard, Peter M. H.; Lavritsen, D.T.;


    ) using immunomagnetic beads (Dynabeads(R)) was developed for isolation of A. pleuropneumoniae serotype 2 from pure cultures and from heterogeneous suspensions. Different coating and washing procedures were evaluated in pure and mixed cultures using polyclonal (PAb) and monoclonal antibodies. The highest...... reisolation yield was achieved when the beads were coated with 1.5 mug PAb IgG/10(7) beads. After washing the beads for four times 9-24% of the bacteria could be reisolated depending on the amount of IgG attached to the beads and the number of beads used. The recovery was increased to 19-61% when only two...... in this heterogeneous suspension. No significant difference was observed when comparing the recovery of A. pleuropneumoniae from pure culture, from mixed cultures and from artificially inoculated tonsils. From 12 pigs inoculated with an aerosol of A. pleuropneumoniae serotype 2 the bacterium could not be detected from...

  8. The genetic organization of the capsular polysaccharide biosynthesis region of Actinobacillus pleuropneumoniae serotype 15

    Ito, Hiroya; Sueyoshi, Masuo


    Nucleotide sequence determination and analysis of the cps gene involved in the capsular polysaccharide biosynthesis of Actinobacillus pleuropneumoniae serotype 15 revealed the presence of three open reading frames, designated as cps15ABC genes. At the protein level, Cps15A and Cps15B showed considerably high homology to CpsA (67.0 to 68.7%) and CpsB (31.7 to 36.8%), respectively, of A. pleuropneumoniae serotypes 1, 4 and 12, revealing the common genetic organization of the cps among serotypes...

  9. Detection of an Actinobacillus pleuropneumoniae serotype 2 lipopolysaccharide (LPS) variant

    Stenbaek, E.I.; HovindHaugen, K.


    linear repeating pentasaccharide units with an O-acetyl group linked to a glucose unit. A monoclonal antibody (MAb 102-G02) directed against A. pleuropneumoniae serotype 2 was characterized in enzyme linked immunosorbent assay (ELISA) and in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS...

  10. Genome biology of Actinobacillus pleuropneumoniae JL03, an isolate of serotype 3 prevalent in China.

    Zhuofei Xu

    Full Text Available Actinobacillus pleuropneumoniae is the etiologic agent of porcine contagious pleuropneumonia, a cause of considerable world wide economic losses in the swine industry. We sequenced the complete genome of A. pleuropneumoniae, JL03, an isolate of serotype 3 prevalent in China. Its genome is a single chromosome of 2,242,062 base pairs containing 2,097 predicted protein-coding sequences, six ribosomal rRNA operons, and 63 tRNA genes. Preliminary analysis of the genomic sequence and the functions of the encoded proteins not only confirmed the present physiological and pathological knowledge but also offered new insights into the metabolic and virulence characteristics of this important pathogen. We identified a full spectrum of genes related to its characteristic chemoheterotrophic catabolism of fermentation and respiration with an incomplete TCA system for anabolism. In addition to confirming the lack of ApxI toxin, identification of a nonsense mutation in apxIVA and a 5'-proximal truncation of the flp operon deleting both its promoter and the flp1flp2tadV genes have provided convincing scenarios for the low virulence property of JL03. Comparative genomic analysis using the available sequences of other serotypes, probable strain (serotype-specific genomic islands related to capsular polysaccharides and lipopolysaccharide O-antigen biosyntheses were identified in JL03, which provides a foundation for future research into the mechanisms of serotypic diversity of A. pleuropneumoniae.

  11. Comparative profiling of the transcriptional response to iron restriction in six serotypes of Actinobacillus pleuropneumoniae with different virulence potential

    Schou, Kirstine Klitgaard; Friis, Carsten; Angen, Øystein;


    receptor of Neisseria meningitidis, a possible virulence factor which contributes to bacterial survival in rats. Conclusions By comparative analysis of gene expression among 6 different serotypes of A. pleuropneumoniae we identified a common set of presumably essential core genes, involved in iron......Background Comparative analysis of gene expression among serotypes within a species can provide valuable information on important differences between related genomes. For the pig lung pathogen Actinobacillus pleuropneumoniae, 15 serotypes with a considerable variation in virulence potential and...... virulence genes. We used a pan-genomic microarray to study the transcriptional response to iron restriction in vitro in six serotypes of A. pleuropneumoniae (1, 2, 3, 5b, 6, and 7), representing at least two levels of virulence. Results In total, 45 genes were significantly (p <0.0001) up-regulated and 67...

  12. Characterization of the omlA gene from different serotypes of Actinobacillus pleuropneumoniae: a new insight into an old approach

    Ciro César Rossi


    Full Text Available The OmlA protein is a virulence factor of Actinobacillus pleuropneumoniae, an important pathogen in pigs. The polymorphisms present in the omlA gene sequence of 15 reference serotypes of A. pleuropneumoniae and non-serotypable isolates were assessed to determine the possible evolutionary relationship among them and to validate the importance of this gene as a molecular marker for the characterization of this bacterium. Divergence among the 15 serotypes of A. pleuropneumoniae probably resulted initially from two major evolutionary events that led to subsequent differentiation into nine groups. This differentiation makes it possible to characterize most of the serotypes by using bionformatics, thereby avoiding problems with immunological cross-reactivity. A conserved α-helix common to all the serotypes was most likely involved in connecting the protein to the outer membrane and acting as a signal peptide. A previously unknown gene duplication was also identified and could contribute to the genetic variability that makes it difficult to serotype some isolates. Our data support the importance of the omlA gene in the biology of A. pleuropneumoniae and provide a new area of research into the OmlA protein.

  13. Binding of Actinobacillus pleuropneumoniae to Phosphatidylethanolamine

    Jeannotte, Marie-Eve; Abul-Milh, Maan; Dubreuil, J. Daniel; Jacques, Mario


    The gram-negative bacterium Actinobacillus pleuropneumoniae is the causative agent of porcine fibrinohemorrhagic necrotizing pleuropneumonia, a disease that causes important economic losses to the swine industry worldwide. In general, the initial step of bacterial colonization is attachment to host cells. The purpose of the present study was to evaluate the binding of A. pleuropneumoniae serotype 1 to phospholipids, which are the major constituents of biological membranes. Phospholipids serve...

  14. Comparative profiling of the transcriptional response to iron restriction in six serotypes of Actinobacillus pleuropneumoniae with different virulence potential

    Angen Øystein


    Full Text Available Abstract Background Comparative analysis of gene expression among serotypes within a species can provide valuable information on important differences between related genomes. For the pig lung pathogen Actinobacillus pleuropneumoniae, 15 serotypes with a considerable variation in virulence potential and immunogenicity have been identified. This serotypic diversity can only partly be explained by amount of capsule and differences in the RTX toxin genes in their genomes. Iron acquisition in vivo is an important bacterial function and in pathogenic bacteria, iron-limitation is often a signal for the induction of virulence genes. We used a pan-genomic microarray to study the transcriptional response to iron restriction in vitro in six serotypes of A. pleuropneumoniae (1, 2, 3, 5b, 6, and 7, representing at least two levels of virulence. Results In total, 45 genes were significantly (p A. pleuropneumoniae was the up-regulation of a putative cirA-like siderophore in all six serotypes. Three genes, recently described in A. pleuropneumoniae as possibly coding for haemoglobin-haptoglobin binding proteins, displayed significant serotype related up-regulation to iron limitation. For all three genes, the expression appeared at its lowest in serotype 3, which is generally considered one of the least virulent serotypes of A. pleuropneumoniae. The three genes share homology with the hmbR haemoglobin receptor of Neisseria meningitidis, a possible virulence factor which contributes to bacterial survival in rats. Conclusions By comparative analysis of gene expression among 6 different serotypes of A. pleuropneumoniae we identified a common set of presumably essential core genes, involved in iron regulation. The results support and expand previous observations concerning the identification of new potential iron acquisition systems in A. pleuropneumoniae, showing that this bacterium has evolved several strategies for scavenging the limited iron resources of the

  15. Evaluation of an enzyme-linked immunosorbent assay for serological surveillance of infection with Actinobacillus pleuropneumoniae serotype 5 in pig herds

    Klausen, Joan; Andresen, Lars Ole; Barfod, Kristen; Sørensen, Vibeke


    An indirect enzyme-linked immunoassay for serological surveillance of infection of pigs with Actinobacillus pleuropneumoniae (Ap) serotype 5 was developed. The antigen used was prepared from Ap serotype 5b strain L20. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis...

  16. Effect of tulathromycin on the carrier status of Actinobacillus pleuropneumoniae serotype 2 in the tonsils of pigs

    Angen, Øystein; Andreasen, M.; Nielsen, E.O.;


    The effect of a single or double dose of tulathromycin was evaluated in pigs carrying Actinobacillus pleuropneumoniae serotype 2 in their tonsils. Twenty-nine pigs from a reinfected specific pathogen-free-herd were selected from animals testing positive in an A pleuropneumoniae serotype 2-specific...... were tested by PCR on tonsil scrapings on days 0, 4, 11 and 33, and on day 33 all the animals were euthanased. There were no significant differences between the numbers of PCR-positive animals in the three groups on any of the sampling dates....... PCR test on tonsil scrapings and they were divided into three groups. The pigs in group I were treated subcutaneously with 2.5 mg/kg tulathromycin on day 0, the pigs in group 2 were treated with 2.5 mg/kg tulathromycin on days 0 and 4, and the pigs in group 3 were left untreated as controls. The pigs...

  17. Evaluation of a multiplex PCR test for simultaneous identification and serotyping of Actinobacillus pleuropneumoniae serotypes 2, 5, and 6

    Jessing, Stine Graakjær; Angen, Øystein; Inzana, Tomas J.


    6 were combined with the already existing species-specific primers used in a PCR test based on the omlA gene. The PCR test was evaluated with serotype reference strains of A. pleuropneumoniae as well as 182 Danish field isolates previously serotyped by latex agglutination or immunodiffusion. For all...... cross-reacted by the latex agglutination test were of serotype 2, 5, or 6. Determination of the serotype by PCR represents a convenient and specific method for the serotyping of A. pleuropneumoniae in diagnostic laboratories....

  18. An atypical biotype I Actinobacillus pleuropneumoniae serotype 13 is present in North America

    Perry, Malcolm B.; Angen, Øystein; MacLean, Leann L.;


    analysis of the capsular polysaccharide (CPS) and lipopolysaccharide (LPS) of a representative strain revealed that the CPS is almost identical to that of the reference strain of serotype 13, having a slightly higher degree of glycose O-acetylation. However, it produces an O-PS within the LPS antigenically...... and structurally identical with that of the reference strain of A. pleuropneumoniae serotype 10. The O-PS was characterized as a homopolymer of 1,2 linked β-d-galactofuranosyl residues, a structure unrelated to that of the O-PS produced by the reference strain of serotype 13. Strains from Canada and...

  19. Serological characterization of Actinobacillus pleuropneumoniae biotype 1 strains antigenically related to both serotypes 2 and 7

    Nielsen, R.; Andresen, Lars Ole; Plambeck, Tamara


    ). Immunodiffusion confirmed the antigenic relationship with serotype 2 and further demonstrated an antigenic relationship with strain WF83 (reference strain of serotype 7). SDS-PAGE with LPS from strains 1536, 4226, WF83 and strain 7317 (representative of the 9 isolates examined) showed that strains WF83 and 7317...

  20. Comparison of high and low virulence serotypes of Actinobacillus pleuropneumoniae by quantitative real-time PCR

    Schou, Kirstine Klitgaard; Angen, Øystein; Boye, Mette

    high virulence while serotype 6 strains are normally found to be less pathogenic. To gain an understanding of the differential virulence of serotype 2 and 6, the expression of a panel of Ap genes during infection of porcine epithelial lung cells (SJPL) were examined by quantitative real-time PCR (qPCR...... be important for early establishment of the bacteria in the host were examined by qPCR. The genes examined were apfA, coding for a subunit of Type IV pili, kdsB coding for a gene involved in lippopolysacceride biosynthesis, and pgaB which is involved in biofilm formation, all three believed to be...... normalization of the qPCR data. Preliminary results showed that in both serotype 2 and serotype 6, the toxin producing gene apxIV was the most highly expressed of the investigated genes. The major difference observed between the two serotypes was that apfA, involved in type IV vili production, was significantly...

  1. Padronização de três ELISAs polivalentes com lipopolissacarídeos de cadeia longa dos sorotipos 1 e 5, 2, 3 e 7 ou 10 e 12 de Actinobacillus pleuropneumoniae Standardization of three polyvalent ELISA based on long chain lipopolysaccharides of serotypes 1 and 5, 2, 3 and 7, or 10 and 12 of Actinobacillus pleuropneumoniae

    S.S. Kuchiishi


    Full Text Available Três ELISAs polivalentes baseados em lipopolissacarídeos de cadeia longa (LPS-CL foram estabelecidos para detectar anticorpos para todos os sorotipos prevalentes de Actinobacillus pleuropneumoniae. Foram testadas amostras provenientes do banco de soros de suínos experimentalmente inoculados com todos os sorotipos de A. pleuropneumoniae. Os ELISAs foram sensíveis à detecção de anticorpos contra todos os LPS-CL. Foram observadas reações cruzadas no ELISA polivalente produzido com os sorotipos 1 e 5, com anti-soros específicos para os sorotipos 9 e 11, pois os sorotipos 1, 9 e 11 apresentaram antígenos somáticos comuns. No polivalente com os sorotipos 2, 3 e 7, observaram-se reações com anti-soros dos sorotipos 4, 6 e 8, devido à presença de antígenos somáticos entre os sorotipos 3, 6 e 8 e entre os sorotipos 4 e 7. Amostras de soros de animais infectados com Mycoplasma hyopneumoniae, Mycoplasma flocculare e Haemophilus parasuis, agentes que acometem o sistema respiratório dos suínos, não apresentaram reações cruzadas com os antígenos baseados em LPS-CL.Three polyvalent ELISA based on long chain lipopolysaccharides (LC-LPS were established to detect all prevalent serotypes of Actinobacillus pleuropneumoniae. Samples from a serum bank of experimentally inoculated animals with all serotypes of A. pleuropneumoniae were tested. Antibodies specific to LC-LPS of each serotype were detected. Cross-reactions were observed in the polyvalent ELISA produced with serotypes 1 and 5, with specific antisera to serotypes 9 and 11 due to common somatic antigens presence in serotypes 1, 9, and 11. In the polyvalent with serotypes 2, 3 and 7 reactions were observed with antisera of serotypes 4, 6, and 8, due to the presence of somatic antigens in serotypes 3, 6, and 8 and serotypes 4 and 7. Experimentally infected animals with respiratory agents of swine Mycoplasma hyopneumoniae, Mycoplasma flocculare, and Haemophilus parasuis did not present

  2. Actinobacillus pleuropneumoniae transmission and clinical outbreaks

    Tobias, T.J.


    Actinobacillus pleuropneumoniae is a bacterium causing respiratory disease and mortality in pigs. Outbreaks of clinical disease occur regularly in pig farms. More knowledge of the epidemiology of the bacteria in pig populations should contribute to the design of more effective measures for preventio

  3. Evaluation of 5 ' nuclease assay for detection of Actinobacillus pleuropneumoniae

    Angen, Øystein; Jensen, J.; Lavritsen, D. T.


    , nonspecific reactions appeared when testing dilutions of DNA templates or pure cultures of A. pleuropneumoniae, as well as when testing tonsil scrapings from specific-pathogen-free herds. The diagnostic sensitivity, as evaluated with 586 tonsil scrapings from animals infected with A. pleuropneumoniae, is the...... equal to 30 must be chosen in order to obtain reliable results. The investigation emphasizes that a thorough evaluation of the criteria used to define a positive test result is necessary.......Sequence detection by the 5' nuclease TaqMan assay uses online detection of internal fluorogenic probes in closed PCR tubes. Primers and probe were chosen from a part of the omlA gene common to all serotypes of Actinobacillus pleuropneumoniae, which gave an amplicon of 92 bp, The test was evaluated...

  4. Mapping of functional regions on the transferrin-binding protein (TfbA) of Actinobacillus pleuropneumoniae.

    Strutzberg, K; von Olleschik, L; Franz, B.; Pyne, C; Schmidt, M. A.; Gerlach, G F


    Actinobacillus pleuropneumoniae can use porcine transferrin as the sole source of iron. Two proteins with molecular masses of approximately 60 kDa (TfbA) and 110 kDa have been shown to specifically bind porcine transferrin; from the TfbA protein, three isoforms from A. pleuropneumoniae serotypes 1, 5, and 7 have been identified and characterized by nucleotide sequence analysis. Here we defined the transferrin-binding region(s) of the TfbA protein of A. pleuropneumoniae serotype 7 by TnphoA mu...

  5. Survival of Actinobacillus pleuropneumoniae outside the pig.

    Assavacheep, P; Rycroft, A N


    Transmission of Actinobacillus pleuropneumoniae is primarily thought to be via direct transfer of mucus from pig to pig. For transfer between farms, the organism may need to persist in the wet or dried state to be carried on an inanimate surface. The survival of A. pleuropneumoniae was investigated under controlled laboratory conditions. In aqueous suspension, survival was improved by the presence of NaCl and mucin; it was prolonged at lower temperature. In dry state, it survived best on a hydrophobic surface either under desiccated conditions or saturated humidity. Detectable viability was maintained for 3-4 days. When frozen, A. pleuropneumoniae survived for more than 17 weeks at -20 °C, but the viability declined to 0.01% during that time. Survival at -70 °C was effective for long term storage. Results obtained from this investigation would be applicable for sampling method, transport techniques, epidemiological study, and biosecurity implementation. PMID:22892250

  6. Evaluation and application of ribotyping for epidemiological studies of Actinobacillus pleuropneumoniae in Denmark

    Fussing, V.; Barfod, Kristen; Nielsen, R.; Møller, K.; Nielsen, J.P.; Wegener, Henrik Caspar; Bisgaard, M.


    The aim of the present study was to evaluate ribotyping as an epidemiological tool for Actinobacillus pleuropneumoniae and apply the method in studies of A. pleuropneumoniae infections in Danish pig herds. The evaluation of ribotyping was based on the 13 international reference strains and 106 ep...... strains were closely related, though only showing 33% similarity to HindIII ribotypes of remaining serotypes. (C) 1998 Elsevier Science B.V. All rights reserved....

  7. Characterization of two genes encoding distinct transferrin-binding proteins in different Actinobacillus pleuropneumoniae isolates.

    Gerlach, G F; Klashinsky, S; Anderson, C.; Potter, A A; Willson, P.J.


    The gene encoding the Actinobacillus pleuropneumoniae serotype 1 transferrin-binding protein (tfbA) was cloned, and the carboxy-terminal 70% of the protein was expressed as an aggregate protein in Escherichia coli. The nucleotide sequences of the tfbA genes from A. pleuropneumoniae serotypes 7 (G.-F. Gerlach, C. Anderson, A. A. Potter, S. Klashinsky, and P. J. Willson, Infect. Immun. 60:892-898, 1992) and 1 were determined, and a comparison revealed that they had 65% sequence identity. The de...

  8. Evaluation of 5 ' nuclease assay for detection of Actinobacillus pleuropneumoniae

    Angen, Øystein; Jensen, J.; Lavritsen, D. T.


    Sequence detection by the 5' nuclease TaqMan assay uses online detection of internal fluorogenic probes in closed PCR tubes. Primers and probe were chosen from a part of the omlA gene common to all serotypes of Actinobacillus pleuropneumoniae, which gave an amplicon of 92 bp, The test was evaluated...... with 73 lung isolates and 120 tonsil isolates of A. pleuropneumoniae as well as with a collection of reference strains. By using a C-t value (cycle number in which the fluorescence exceeds the threshold defined by the software) of 30 as the cutoff limit, the 5' nuclease assay represents a test with 100......% sensitivity and 100% specificity, A high degree of reproducibility of the test was demonstrated. If samples with C-t values of less than or equal to 30 are considered positive, the detection limit of the assay was 1 CPU/reaction tube, corresponding to a 10-fold higher number of DNA templates. After cycle 30...

  9. Estudios hematológicos y patológicos comparativos de cerdos inoculados con un aislado de campo y el serotipo 5 ATCC de Actinobacillus pleuropneumoniae Comparative hematological and pathological study of inoculated pigs with a field isolate and an ATCC serotype 5 of Actinobacillus pleuropneumoniae

    D Muñoz


    Full Text Available Se realizó una inoculación experimental de A. pleuropneumoniae utilizando un aislado de campo y una cepa de referencia ATCC serotipo 5, para lo cual se utilizaron tres grupos de animales (n = 15 para cada grupo. El grupo 1 (G1 fue inoculado con medio estéril, el grupo (G2 con serotipo 5 ATCC y el grupo 3 (G3 fue inoculado con un aislado de campo (418/07. Los resultados mostraron diferencias significativas (P ≤ 0,05 en el recuento de leucocitos totales entre el grupo G1 v/s G2 y G1 v/s G3 y los grados de las lesiones pulmonares totales evidenciaron diferencias estadísticamente significativas (P ≤ 0,05 entre los tres grupos de estudio. Las lesiones histopatológicas pulmonares mostraron diferencias estadísticas relevantes sólo entre G1 y G3 (P ≤ 0,05. En este trabajo se verifican diferencias importantes del comportamiento entre el aislado de campo y el serotipo 5 ATCC, sobre los cambios hematológicos y las lesiones macroscópicas e histopatológicas ocasionadas por ellos, lo cual podría indicar una mayor virulencia y patogenicidad del aislado nacional. Se espera en un futuro próximo serotipificar este aislado nacional de App.An experimental inoculation of Actinobacillus pleuropneumoniae (App was carried out with a field isolate and an ATCC serotype 5. Three groups of 15 pigs each were used. Group 1 (G1 was the control group inoculated with sterile media, Group 2 was inoculated with the serotype 5 ATCC, and Group 3 (G3 was inoculated with a field isolate (418/07. The results showed statistically significant differences (P ≤ 0.05 in the total leukocytes count between G1 v/s G2 and G1 v/s G3. The total macroscopic lung lesions scores were statistically different among the 3 groups (P ≤ 0.05. However, statistical difference was found only between G1 and G3 in the histopathological lung lesions (P ≤ 0.05. This work shows a clear difference in the hematological changes and the macroscopic and histopathological lesions between the

  10. A cohort study on Actinobacillus pleuropneumoniae colonisation in suckling piglets

    Tobias, T.J.; Klinkenberg, D.; Bouma, A.; Broek, van den J.; Daemen, A.J.J.M.; Wagenaar, J.A.; Stegeman, J.A.


    Actinobacillus pleuropneumoniae causes respiratory disease in pigs and despite the use of preventive measures such as vaccination and antimicrobials clinical outbreaks still occur. At weaning often many piglets are not colonised. If differences in prevalence between litters are large and if factors

  11. Improved diagnostic PCR assay for Actinobacillus pleuropneumoniae based on the nucleotide sequence of an outer membrane lipoprotein

    Gram, Trine; Ahrens, Peter


    species related to A. pleuropneumoniae or isolated from pigs were assayed. They were all found negative in the PCR, as were tonsil cultures from 50 pigs of an A. pleuropneumoniae-negative herd. The sensitivity assessed by agarose gel analysis of the PCR product was 10(2) CFU/PCR test tube. The specificity......The gene (omlA) coding for an outer membrane protein of Actinobacillus pleuropneumoniae serotypes 1 and 5 has been described earlier and has formed the basis for development of a specific PCR assay, The corresponding regions of all 12 A. pleuropneumoniae reference strains of biovar 1 were sequenced...... and sensitivity of this PCR compared to those of culture suggest the use of this PCR for routine identification of A. pleuropneumoniae....

  12. Transmission of Actinobacillus pleuropneumoniae in pigs under field-like conditions: emphasis on tonsillar colonisation and passively acquired colostral antibodies

    Vigre, Håkan; Angen, Øystein; Barfod, K.; Lavritsen, D. T.; Sørensen, V.

    proportion of pigs with detectable levels of colostral antibodies to the different serotypes of A. pleuropneumoniae was declining. Since these two events take place in the same age period, we expect a possible biological association between the level of the passive immunity and the degree of tonsillar......The objectives of this study were to elucidate at which age tonsillar colonisation by Actinobacillus pleuropneumoniae occurs in pigs and relate this occurrence to the presence of colostral antibodies to A. pleuropneumoniae. The infection patterns were studied in an isolated cohort of pigs, which...... birth, showing that A. pleuropneumoniae can be transmitted from sow to offspring during a 3-week nursing period. The cumulative proportion of pigs carrying A. pleuropneumoniae in their tonsils increased significantly between the age of 4-12 weeks. This age period corresponded to the age at which the...

  13. Isolation and identification of Actinobacillus pleuropneumoniae in pig's lungs at farms and their sensitivity to antibiotics

    Žutić Milenko; Ašanin Ružica; Milić N.; Ivetić V.; Vidić Branka; Žutić Jadranka; Ašanin Jelena


    The presence of Actinobacillus pleuropneumoniae has been established in all suspected cases of pleuropneumonia at several farms that have been included in the research. Equal incidence of pleuropneumonia has been found both among piglets for breeding and for fattening. The health monitoring of herds is extremely important, firstly because of the need for the adequate strategy to be chosen for controlling the Actinobacillus-caused pleuropneumonia and, at the same time, in order to prevent enor...

  14. Catecholamines promote Actinobacillus pleuropneumoniae growth by regulating iron metabolism.

    Lu Li

    Full Text Available Catecholamines are host stress hormones that can induce the growth of many bacteria by facilitating iron utilization and/or regulate the expression of virulence genes through specific hormone receptors. Whether these two responsive pathways are interconnected is unknown. In our previous study, it was found that catecholamines can regulate the expression of a great number of genes of Actinobacillus pleuropneumoniae, an important swine respiratory pathogen. However, bacterial growth was not affected by catecholamines in rich medium. In this study, it was discovered that catecholamines affected A. pleuropneumoniae growth in chemically defined medium (CDM. We found that serum inhibited A. pleuropneumoniae growth in CDM, while epinephrine, norepinephrine and dopamine promoted A. pleuropneumoniae growth in the CDM containing serum. The known bacterial hormone receptor QseC didn't play roles in this process. Ion-supplementation and transcriptome analysis indicated that serum addition resulted in iron-restricted conditions which were alleviated by the addition of catecholamines. Transferrin, one of the components in serum, inhibited the growth of A. pleuropneumoniae in CDM, an effect reversed by addition of catecholamines in a TonB2-dependent manner. Our data demonstrate that catecholamines promote A. pleuropneumoniae growth by regulating iron-acquisition and metabolism, which is independent of the adrenergic receptor QseC.

  15. Proteomic and immunoproteomic characterization of a DIVA subunit vaccine against Actinobacillus pleuropneumoniae

    Maas Alexander


    Full Text Available Abstract Background Protection of pigs by vaccination against Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is hampered by the presence of 15 different serotypes. A DIVA subunit vaccine comprised of detergent-released proteins from A. pleuropneumoniae serotypes 1, 2 and 5 has been developed and shown to protect pigs from clinical symptoms upon homologous and heterologous challenge. This vaccine has not been characterized in-depth so far. Thus we performed i mass spectrometry in order to identify the exact protein content of the vaccine and ii cross-serotype 2-D immunoblotting in order to discover cross-reactive antigens. By these approaches we expected to gain results enabling us to argue about the reasons for the efficacy of the analyzed vaccine. Results We identified 75 different proteins in the vaccine. Using the PSORTb algorithm these proteins were classified according to their cellular localization. Highly enriched proteins are outer membrane-associated lipoproteins like OmlA and TbpB, integral outer membrane proteins like FrpB, TbpA, OmpA1, OmpA2, HgbA and OmpP2, and secreted Apx toxins. The subunit vaccine also contained large amounts of the ApxIVA toxin so far thought to be expressed only during infection. Applying two-dimensional difference gel electrophoresis (2-D DIGE we showed different isoforms and variations in expression levels of several proteins among the strains used for vaccine production. For detection of cross-reactive antigens we used detergent released proteins of serotype 7. Sera of pigs vaccinated with the detergent-released proteins of serotypes 1, 2, and 5 detected seven different proteins of serotype 7, and convalescent sera of pigs surviving experimental infection with serotype 7 reacted with 13 different proteins of the detergent-released proteins of A. pleuropneumoniae serotypes 1, 2, and 5. Conclusions A detergent extraction-based subunit vaccine of A. pleuropneumoniae was

  16. Interference of peptides and specific antibodies with the function of the Actinobacillus pleuropneumoniae transferrin-binding protein.

    Strutzberg, K; Franz, B.; Gerlach, G F


    Multiple-antigenic peptides (MAPs) containing transferrin-binding domains of the Actinobacillus pleuropneumoniae serotype 7-derived transferrin-binding protein (TfbA) (K. Strutzberg, L. von Olleschik, B. Franz, C. Pyne, M. A. Schmidt, and G.-F. Gerlach, Infect. Immun. 63:3846-3850, 1995) were constructed. It was found that the MAPs inhibited transferrin binding of the recombinant TfbA protein, whereas antibodies directed against transferrin-binding domains failed to do so.

  17. Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to the Apx toxins of Actinobacillus pleuropneumoniae

    Nielsen, Ragnhild; van den Bosch, Johannes F.; Plambeck, Tamara; Sørensen, Vibeke; Nielsen, Jens Peter

    The reference strains of the 12 serotypes of Actinobacillus pleuropneumoniae express one or two of three different RTX exotoxins designated Apr I, Apr II and Apr III. The toxins are important virulence factors. In the present study, ELISAs with purified Apr I, Apr II and Apr III, respectively, as...

  18. Transcriptional profiling of Actinobacillus pleuropneumoniae during the acute phase of a natural infection in pigs

    Harel Josée


    Full Text Available Abstract Background Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia, a respiratory disease which causes great economic losses worldwide. Many virulence factors are involved in the pathogenesis, namely capsular polysaccharides, RTX toxins, LPS and many iron acquisition systems. In order to identify genes that are expressed in vivo during a natural infection, we undertook transcript profiling experiments with an A. pleuropneumoniae DNA microarray, after recovery of bacterial mRNAs from serotype 5b-infected porcine lungs. AppChip2 contains 2033 PCR amplicons based on the genomic sequence of App serotype 5b strain L20, representing more than 95% of ORFs greater than 160 bp in length. Results Transcriptional profiling of A. pleuropneumoniae recovered from the lung of a pig suffering from a natural infection or following growth of the bacterial isolate in BHI medium was performed. An RNA extraction protocol combining beadbeating and hot-acid-phenol was developed in order to maximize bacterial mRNA yields and quality following total RNA extraction from lung lesions. Nearly all A. pleuropneumoniae transcripts could be detected on our microarrays, and 150 genes were deemed differentially expressed in vivo during the acute phase of the infection. Our results indicate that, for example, gene apxIVA from an operon coding for RTX toxin ApxIV is highly up-regulated in vivo, and that two genes from the operon coding for type IV fimbriae (APL_0878 and APL_0879 were also up-regulated. These transcriptional profiling data, combined with previous comparative genomic hybridizations performed by our group, revealed that 66 out of the 72 up-regulated genes are conserved amongst all serotypes and that 3 of them code for products that are predicted outer membrane proteins (genes irp and APL_0959, predicted to code for a TonB-dependent receptor and a filamentous hemagglutinin/adhesin respectively or lipoproteins (gene APL_0920. Only 4

  19. Concurrent host-pathogen gene expression in the lungs of pigs challenged with Actinobacillus pleuropneumoniae

    Brogaard, Louise; Schou, Kirstine Klitgaard; Heegaard, Peter M. H.;


    Background: Actinobacillus pleuropneumoniae causes pleuropneumonia in pigs, a disease which is associated with high morbidity and mortality, as well as impaired animal welfare. To obtain in-depth understanding of this infection, the interplay between virulence factors of the pathogen and defense ...

  20. Purification, serology and pathogenic role of the 110 kilodalton rtx hemolysins of Actinobacillus pleuropneumoniae

    Ma, Jianneng


    Actinobacillus pleuropneumoniae is the etiological agent of contagious swine pleuropneumonia, an economically important disease of the swine industry worldwide. Improved control of this disease requires enhanced understanding of the factors contributing to pathogenesis. The objectives of this study were to investigate the immune response and virulence properties of the 110-kilodalton (110-KDa) hemolysins [hemolysin I (HlyI) and hemolysin II (HlyII)] of A. pleuropneumoniae...

  1. No overall relationship between average daily weight gain and the serological response to Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae in eight chronically infected Danish swine herds

    Andreasen, Margit; Mousing, Jan; Thomsen, Lars Krogsgård

    approximately 4 weeks of age), and sera were analyzed for antibodies to Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae serotypes 2, 5-7 and 12. Mixed analysis of covariance analyzed the relationship between the average daily weight gain and a categorical variable defining seroconversion as none...... most pigs included in the study were subclinically infected, or because a temporary negative influence of the infections is hidden due to an increased growth in the period following infection. In conclusion. at least in these eight herds, seroresponses to M. hyopneumoniae and A. pleuropneumoniae could...

  2. Characterization of an Actinobacillus pleuropneumoniae seeder pig challenge-exposure model.

    Lechtenberg, K F; Shryock, T R; Moore, G


    Five strains of Actinobacillus pleuropneumoniae serotype 1 were used to intranasally infect 5 groups of pigs. Using each bacterial strain, infected pigs (termed seeder pigs) were commingled for 48 hours with 5 groups of noninfected test pigs, then were removed. Seeder and test pigs were maintained in isolation and were observed for 14 days. Seeder pigs had mortality that was threefold greater than that of test pigs (24% vs 8%). Rectal temperature in excess of 40.3 C was achieved for 84% of test pigs and 88% of seeder pigs. Neither of these 2 variables was statistically different between the 2 groups of pigs. Clinical impression scores > or = 2 (on a 0 to 3 scale) were three-fold (64% vs 20%) greater for seeder than for test pigs (P amount of lesions. The number of A pleuropneumoniae isolations was not statistically different between test and seeder pig populations. Recovery of Pasteurella multocida or other bacteria was greater from the seeder pigs (P recoverable isolates was greater from test pigs than from seeder pigs (P < 0.05). Assessment of lung lesions at necropsy by either visual estimation or on a weight basis were in agreement.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7887514

  3. Effects of Actinobacillus pleuropneumoniae cytotoxins on generation of oxygen radicals by porcine neutrophils

    Simson Tarigan


    Full Text Available Cytotoxins produced by Actinobacillus pleuropneumoniae (App suggested to be the most important pathogenic and virulent factors for this organism. However, the mechanisms on how the cytotoxins contribute to the disease process remain unclear. The purpose of this study is to investigate the effect of the cytotoxins on the oxidative-burst metabolism of porcine neutrophils. In this study, neutrophils were firstly loaded with an oxidative probe dichlorofluorescin diacetate (DCFHDA then expose to cytotoxins. Cells producing oxygen radicals emitted fluorescence and its intensity was measured with a FACScan flow cytometer. All cytotoxins derived from either App serotypes producing ApxI and ApxII, App serotypes producing ApxII only, or App serotypes producing ApxII and ApxIII were capable of stimulating neutrophils for oxygen-radical generation. However, compared with phorbol myristate acetate (PMA, App cytotoxins were much weaker as stimulants for oxygen radicals. In addition, Apx preparation stimulated an oxidative-burst metabolism of neutrophils at a low, narrow range of Apx doses. At higher doses, the toxins inhibit the oxidative burst metabolism. The effects of cytotoxins produced by App during infection on recruited neutrophils into the lungs are assumed to be comparable to those observed in this in vitro study. Neutrophils, and other host cells, adjacent to the bacteria become lysis due to high toxin concentration, whereas those at some distance to the bacteria produce oxygen radicals which in turn cause tissue damage or necrosis.

  4. Cloning and Expression of Actinobacillus pleuropneumoniae Gene Coding for TbpA and Development of an Indirect TbpA-ELISA

    LIANG Wang-wang; HE Qi-gai; CHEN Huan-chun; XU Di-ping; WU Rui; ZHANG Rong-rong


    This study presents the cloning and expression of gene encoding transferrin-binding protein A from Actinobacillus pleuropneumoniae in Escherichia coli expression system and the development of an indirect TbpA-ELISA. The gene coding TbpA was amplified from the A. pleuropneumoniae serotype 2 genome using polymerase chain reaction and cloned to pET-28b expression vector under the control of strong, inducible T7 promoter. The recombinant plasmid was expressed in E. coli BL21 (DE3). The expressed fusion protein was analyzed using SDS-PAGE and Western blotting. The diagnostic potential of recombinant TbpA (rTbpA) was evaluated through an antibody-detection indirect ELISA based on the purified rTbpA. The TbpA antibodies were detectable in mice on day 7 after vaccination with purified rTbpA protein or infection with A. pleuropneumoniae serotype 10 with the TbpA-based ELISA. In addition, the TbpA-ELISA was able to detect 12 serotyping rabbit antisera postinoculation (PI) with A. pleuropneumoniae 12 serotypes experimentally. The comparable result was obtained by detecting the 117 clinical serum samples using, respectively, the TbpA-ELISA and indirect hemagglutination test (IHA) based on multiplex antigen. The result indicates that the TbpA-ELISA was the more sensitive method compared with the Mix-IHA method because of its consistent presence in A. pleuropneumoniae serotypes. In conclusion, the conserved TbpA of A. pleuropneumoniae can be used for the development of a cross-serotype diagnostic method for the detection of antibodies against A. pleuropneumoniae.

  5. Effect of different antimicrobial treatments on serum acute phase responses and leukocyte counts in pigs after a primary and a secondary challenge infection with Actinobacillus pleuropneumoniae

    Sjölund, M; Fossum, C; Martin de la Fuente, AJM;


    The susceptibility to an initial challenge and a re-challenge inoculation with Actinobacillus pleuropneumoniae was analysed in pigs that were treated with antimicrobials of different efficacies following the first exposure to A pleuropneumoniae. In brief, 30 nine-week-old specific pathogen......-free pigs were allocated to five groups of six. After acclimatisation, four groups were inoculated with A pleuropneumoniae serotype 2. At the onset of clinical signs, three of the groups of pigs were treated with enrofloxacin, tetracycline or penicillin. A fourth group served as the inoculated control and...

  6. Diversidad genética de cepas de Actinobacillus pleuropneumoniae (App aisladas desde planteles de producción intensiva de cerdos en Chile Genetic diversity of Actinobacillus pleuropneumoniae (App strains in intensive swine farms in Chile

    V Neira-Ramírez


    Full Text Available Actinobacillus pleuropneumoniae (App es el agente etiológico de la pleuroneumonía contagiosa porcina, una de las enfermedades de etiología bacteriana de mayor relevancia en producción porcina. En el mundo se han descrito 15 serotipos de App, en Chile solo los serotipos 1 y 5. La serotipificación requiere mucho tiempo, trabajo y dinero, actualmente se encuentran herramientas moleculares para realizar una "serotipificación" mediante la genotipificación de toxinas Apx. Así, se evaluaron 60 aislados de App provenientes de nueve empresas porcinas de producción intensiva distribuidas en distintas regiones de Chile, obtenidas desde pulmones de cerdos con lesiones compatibles con pleuroneumonía contagiosa porcina. Las bacterias fueron aisladas mediante los métodos tradicionales y confirmados por API, recolectados durante los años 2007, 2008 y 2009. Los resultados identificaron los genotipos correspondientes sólo a los serotipos 4, 6 y 7, los cuales se describen por primera vez en Chile, siendo el más frecuente el serotipo 7. En las diferentes zonas estudiadas, no existió un serotipo predominante, excepto en las regiones de O'Higgins y del Biobío en las cuales fue más frecuentemente aislado el serotipo 7. El presente estudio es el primer acercamiento con el fin de conocer la distribución de serotipos de App en Chile. Con el fin de conocer la real diversidad genética y serotipos de App en los diversos planteles en Chile es necesario realizar estudios que contemplen un mayor número de aislados.Actinobacillus pleuropneumoniae (App is the etiologic agent of porcine contagious pleuropneumonia, an important bacterial disease in intensive pig production. In the world were described 15 App serotypes, in Chile serotypes 1 and 5 have been reported. The serotyping technique is slow, expensive and difficult; currently, a molecular tool named PCR is available to "serotyping" by Apx toxins genotyping, which is quick, non-expensive and easy. 60 App

  7. Multiplex PCR that can distinguish between immunologically cross-reactive serovar 3, 6, and 8 Actinobacillus pleuropneumoniae strains

    Zhou, L.; Jones, S.C.P.; Angen, Øystein; Bosse, J.T.; Nash, J. H. E.; Frey, J.; Zhou, R.; Chen, H.C.; Kroll, J.S.; Rycroft, A.N.; Langford, P.R.


    We describe a highly sensitive and specific multiplex PCR, based on capsular loci and the species specific apxIV gene, that unequivocally differentiates serovar 3, 6, and 8 Actinobacillus pleuropneumoniae strains that are cross-reactive in conventional immunological tests.......We describe a highly sensitive and specific multiplex PCR, based on capsular loci and the species specific apxIV gene, that unequivocally differentiates serovar 3, 6, and 8 Actinobacillus pleuropneumoniae strains that are cross-reactive in conventional immunological tests....

  8. Influence of Serum and Glucose Additives on Survival of Actinobacillus pleuropneumoniae Aerosolized from the Freeze-Dried State

    Hensel, Andreas


    Serum and/or glucose added to Actinobacillus pleuropneumoniae suspensions before freeze-drying significantly increased survival rates of bacteria in aerosols. Aerosols with predictable numbers of viable bacteria can be made as required in an aerosol infection model. Sucrose supplementation of impinger fluids increased recovery of viable A. pleuropneumoniae.

  9. Cloning and molecular characterization of Cu,Zn superoxide dismutase from Actinobacillus pleuropneumoniae.

    Langford, P R; Loynds, B M; Kroll, J S


    Copper-zinc superoxide dismutases (Cu,Zn SODs), until recently considered very unusual in bacteria, are now being found in a wide range of gram-negative bacterial species. Here we report the cloning and characterization of sodC, encoding Cu,Zn SOD in Actinobacillus pleuropneumoniae, a major pathogen of pigs and the causative organism of porcine pleuropneumonia. sodC was shown to lie on a monocistronic operon, at the chromosomal locus between the genes asd (encoding aspartate semialdehyde dehy...

  10. Profiling microRNAs in lung tissue from pigs infected with Actinobacillus pleuropneumoniae

    Podolska, Agnieszka; Anthon, Christian; Bak, Mads; Tommerup, Niels; Skovgaard, Kerstin; Heegaard, Peter M. H.; Gorodkin, Jan; Cirera, Susanna; Fredholm, Merete


    still very limited. Results: In this study, the RNA extracted from visually unaffected and necrotic tissue from pigs infected with Actinobacillus pleuropneumoniae was subjected to small RNA deep sequencing. We identified 169 conserved and 11 candidate novel microRNAs in the pig. Of these, 17 were......Background: MicroRNAs (miRNAs) are a class of non-protein-coding genes that play a crucial regulatory role in mammalian development and disease. Whereas a large number of miRNAs have been annotated at the structural level during the latest years, functional annotation is sparse. Actinobacillus...... pleuropneumoniae (APP) causes serious lung infections in pigs. Severe damage to the lungs, in many cases deadly, is caused by toxins released by the bacterium and to some degree by host mediated tissue damage. However, understanding of the role of microRNAs in the course of this infectious disease in porcine is...

  11. Evaluation of a PCR for detection of Actinobacillus pleuropneumoniae in mixed bacterial cultures from tonsils

    Gram, T.; Ahrens, Peter; Nielsen, J.P.


    strains of A. lignieresii. The lower detection limit of the PCR test was 10(3) A. pleuropneumoniae CFU/PCR test tube and was not affected by addition of 10(6) E. coli CFU/PCR test tube. Mixed bacterial cultures from tonsils of 101 pigs from 9 different herds were tested by culture and by PCR using four...... different bacteriological media. While 65% reacted positive; in the PCR only 23% were positive by culture, thereby suggesting a superior sensitivity of the PCR test to that of culture. The use of selective media, large inoculum and incubation for 48 h gave the highest number of positive PCR reactions from......A PCR for the detection of Actinobacillus pleuropneumoniae was evaluated. All of 102 field isolates of A. pleuropneumoniae reacted in the PCR by amplification of a 985 bp product. No PCR amplification product was observed when examining strains of A. ureae, A. capsulatus, A. hominis, A. equuli, A...

  12. Comparison of conventional and long-acting oxytetracyclines in prevention of induced Actinobacillus (Haemophilus) pleuropneumoniae infection of growing swine.

    Kiorpes, A L; Bäckström, L R; Collins, M T; Kruse, G O


    These experiments tested the hypothesis that long-acting oxytetracycline (oxytetracycline-LA) was more effective than regular oxytetracycline in preventing porcine pleuropneumonia when administered either 24 or 48 h prior to experimental challenge with virulent strains of Actinobacillus pleuropneumoniae. Two experiments (1 and 2) were conducted using growing pigs (average weight 12-15 kg). Antibiotic treatments were administered once intramuscularly at 20 mg/kg body weight; controls received ...

  13. A multiplexed immunoassay for detection of antibodies to Actinobacillus pleuropneumoniae (App) in pigs

    Berger, Sanne Schou; Boas, Ulrik; Andresen, Lars Ole;


    The bacterium Actinobacillus pleuropneumoniae (App) is the causative agent of porcine pleuropneumoniae, a contagious and severe respiratory disease in pigs. Based on capsular antigens, 15 App serovars have been described, and the prevalence and morbidity of these serovars vary with geographic...... regions (1). In Denmark, the most important serovars are considered to be App 1, 2, 5, 6, 7, 10 and 12. As part of the Danish surveillance program for App, the Danish Veterinary Institute uses ELISAs and complement fixation tests (CFT) to test for porcine anti-App antibodies (2-7). In an effort to improve...... our diagnostic tools, we are currently developing a novel indirect fluorescent microsphere immunoassay that can facilitate simultaneous detection of antibodies towards multiple App serovars within a single serum sample volume. The multiplex immunoassay is based on Luminex technology (8) and has...

  14. A computational strategy for the search of regulatory small RNAs in Actinobacillus pleuropneumoniae

    Rossi, Ciro C.; Bossé, Janine T.; Li, Yanwen; Witney, Adam A.; Gould, Kate A.; Langford, Paul R.; Bazzolli, Denise M.S.


    Bacterial regulatory small RNAs (sRNAs) play important roles in gene regulation and are frequently connected to the expression of virulence factors in diverse bacteria. Only a few sRNAs have been described for Pasteurellaceae pathogens and no in-depth analysis of sRNAs has been described for Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, responsible for considerable losses in the swine industry. To search for sRNAs in A. pleuropneumoniae, we developed a strategy for the computational analysis of the bacterial genome by using four algorithms with different approaches, followed by experimental validation. The coding strand and expression of 17 out of 23 RNA candidates were confirmed by Northern blotting, RT-PCR, and RNA sequencing. Among them, two are likely riboswitches, three are housekeeping regulatory RNAs, two are the widely studied GcvB and 6S sRNAs, and 10 are putative novel trans-acting sRNAs, never before described for any bacteria. The latter group has several potential mRNA targets, many of which are involved with virulence, stress resistance, or metabolism, and connect the sRNAs in a complex gene regulatory network. The sRNAs identified are well conserved among the Pasteurellaceae that are evolutionarily closer to A. pleuropneumoniae and/or share the same host. Our results show that the combination of newly developed computational programs can be successfully utilized for the discovery of novel sRNAs and indicate an intricate system of gene regulation through sRNAs in A. pleuropneumoniae and in other Pasteurellaceae, thus providing clues for novel aspects of virulence that will be explored in further studies. PMID:27402897

  15. A Transcriptome Map of Actinobacillus pleuropneumoniae at Single-Nucleotide Resolution Using Deep RNA-Seq

    Su, Zhipeng; Zhu, Jiawen; Xu, Zhuofei; Xiao, Ran; Zhou, Rui; Li, Lu; Chen, Huanchun


    Actinobacillus pleuropneumoniae is the pathogen of porcine contagious pleuropneumoniae, a highly contagious respiratory disease of swine. Although the genome of A. pleuropneumoniae was sequenced several years ago, limited information is available on the genome-wide transcriptional analysis to accurately annotate the gene structures and regulatory elements. High-throughput RNA sequencing (RNA-seq) has been applied to study the transcriptional landscape of bacteria, which can efficiently and accurately identify gene expression regions and unknown transcriptional units, especially small non-coding RNAs (sRNAs), UTRs and regulatory regions. The aim of this study is to comprehensively analyze the transcriptome of A. pleuropneumoniae by RNA-seq in order to improve the existing genome annotation and promote our understanding of A. pleuropneumoniae gene structures and RNA-based regulation. In this study, we utilized RNA-seq to construct a single nucleotide resolution transcriptome map of A. pleuropneumoniae. More than 3.8 million high-quality reads (average length ~90 bp) from a cDNA library were generated and aligned to the reference genome. We identified 32 open reading frames encoding novel proteins that were mis-annotated in the previous genome annotations. The start sites for 35 genes based on the current genome annotation were corrected. Furthermore, 51 sRNAs in the A. pleuropneumoniae genome were discovered, of which 40 sRNAs were never reported in previous studies. The transcriptome map also enabled visualization of 5'- and 3'-UTR regions, in which contained 11 sRNAs. In addition, 351 operons covering 1230 genes throughout the whole genome were identified. The RNA-Seq based transcriptome map validated annotated genes and corrected annotations of open reading frames in the genome, and led to the identification of many functional elements (e.g. regions encoding novel proteins, non-coding sRNAs and operon structures). The transcriptional units described in this study

  16. Identification of a hemolysin from Actinobacillus pleuropneumoniae and characterization of its channel properties in planar phospholipid bilayers.

    Lalonde, G; McDonald, T V; Gardner, P; O'Hanley, P D


    A proteinaceous hemolysin secreted by strain 4074 of serotype 1 of Actinobacillus pleuropneumoniae was purified by diafiltration and ion exchange chromatographic techniques. The hemolytic activity is associated with a 107-kDa band as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and confirmed by Western blotting and immunoprecipitation. This hemolysin produces pores in membranes as demonstrated by osmotic protection studies using red blood cells and carbohydrate compounds of various molecular weights. These assays suggest a pore diameter in the order of 2 nm. Phospholipid bilayers composed of 1:1 w/w phosphotidylserine:phosphotidylethanolamine exposed to this toxin display discrete current flow events typical of transmembrane channels and consistent with the interpretation that this toxin acts by forming pores in phospholipid membranes. The linear relationship of current amplitude to holding potential when examined over the -60 to +60 mV range indicates that this pore has a constant mean single channel conductance level of 350-400 pS. PMID:2474533

  17. The transferrin receptor of Actinobacillus pleuropneumoniae: Quantitation of expression and structural characterization using a peptide-specific monoclonal antibody

    Bøg, Yang S.; Andresen, Lars Ole; Bastholm, L.;


    antigens were detected with the Mab in iron-starved Actinobacillus lignieresii, Actinobacillus porcinus, Actinobacillus minor Haemophilus influenzae. and Haemophilus parasuis. Using an enzyme-linked immunosorbent assay (ELISA) based on the Mab 1.48, Tbp2 could be detected in both recombinant E. coli......-restricted culture. It was found that Tbp2 was not expressed in iron replete medium by any serotype except serotypes 5a, 5b and 6 where a weak expression was seen. There was a weak expression of related antigens in Actinobacillus indolicus and Actinobacillus suis under iron-depleted conditions while no similar...... expressing Tbp2 and in wild type A. pp grown under iron restricted conditions. The subcellular location of Tbp2 in A. pp was studied by immunoelectron microscopy using the Mab 1.48. Interestingly, all antibody binding was found inside the A. pp cells, while Tbp2 expressed in recombinant E. coli was found...

  18. Characterization of bifunctional L-glutathione synthetases from Actinobacillus pleuropneumoniae and Actinobacillus succinogenes for efficient glutathione biosynthesis.

    Yang, Jianhua; Li, Wei; Wang, Dezheng; Wu, Hui; Li, Zhimin; Ye, Qin


    Glutathione (GSH), an important bioactive substance, is widely applied in pharmaceutical and food industries. In this work, two bifunctional L-glutathione synthetases (GshF) from Actinobacillus pleuropneumoniae (GshFAp) and Actinobacillus succinogenes (GshFAs) were successfully expressed in Escherichia coli BL-21(DE3). Similar to the GshF from Streptococcus thermophilus (GshFSt), GshFAp and GshFAs can be applied for high titer GSH production because they are less sensitive to end-product inhibition (Ki values 33 and 43 mM, respectively). The active catalytic forms of GshFAs and GshFAp are dimers, consistent with those of GshFPm (GshF from Pasteurella multocida) and GshFSa (GshF from Streptococcus agalactiae), but are different from GshFSt (GshF from S. thermophilus) which is an active monomer. The analysis of the protein sequences and three dimensional structures of GshFs suggested that the binding sites of GshFs for substrates, L-cysteine, L-glutamate, γ-glutamylcysteine, adenosine-triphosphate, and glycine are highly conserved with only very few differences. With sufficient supply of the precursors, the recombinant strains BL-21(DE3)/pET28a-gshFas and BL-21(DE3)/pET28a-gshFap were able to produce 36.6 and 34.1 mM GSH, with the molar yield of 0.92 and 0.85 mol/mol, respectively, based on the added L-cysteine. The results showed that GshFAp and GshFAs are potentially good candidates for industrial GSH production. PMID:26996628

  19. Intra-unit correlations in seroconversion to Actinobacillus pleuropneumoniae and Mycoplasma hyopneumoniae at different levels in Danish multi-site pig production facilities

    Vigre, Håkan; Dohoo, I.R.; Stryhn, H.;


    2) and Mycoplasma hyopneumoniae (Mh). Based on the estimated variances, three newly described computational methods (model linearisation, simulation and linear modelling) and the standard method (latent-variable approach) were used to estimate the correlations (intra-class correlation components......, ICCs) between pigs in the same production unit regarding seroconversion. Substantially different values of ICCs were obtained from the four methods. However, ICCs obtained by the simulation and the model linearisation were quite consistent. Data used for estimation were collected from 1161 pigs from......In this paper, multilevel logistic models which take into account the multilevel structure of multi-site pig production were used to estimate the variances between pigs produced in Danish multi-site pig production facilities regarding seroconversion to Actinobacillus pleuropneumoniae serotype 2 (Ap...

  20. Profiling microRNAs in lung tissue from pigs infected with Actinobacillus pleuropneumoniae

    Podolska Agnieszka


    Full Text Available Abstract Background MicroRNAs (miRNAs are a class of non-protein-coding genes that play a crucial regulatory role in mammalian development and disease. Whereas a large number of miRNAs have been annotated at the structural level during the latest years, functional annotation is sparse. Actinobacillus pleuropneumoniae (APP causes serious lung infections in pigs. Severe damage to the lungs, in many cases deadly, is caused by toxins released by the bacterium and to some degree by host mediated tissue damage. However, understanding of the role of microRNAs in the course of this infectious disease in porcine is still very limited. Results In this study, the RNA extracted from visually unaffected and necrotic tissue from pigs infected with Actinobacillus pleuropneumoniae was subjected to small RNA deep sequencing. We identified 169 conserved and 11 candidate novel microRNAs in the pig. Of these, 17 were significantly up-regulated in the necrotic sample and 12 were down-regulated. The expression analysis of a number of candidates revealed microRNAs of potential importance in the innate immune response. MiR-155, a known key player in inflammation, was found expressed in both samples. Moreover, miR-664-5p, miR-451 and miR-15a appear as very promising candidates for microRNAs involved in response to pathogen infection. Conclusions This is the first study revealing significant differences in composition and expression profiles of miRNAs in lungs infected with a bacterial pathogen. Our results extend annotation of microRNA in pig and provide insight into the role of a number of microRNAs in regulation of bacteria induced immune and inflammatory response in porcine lung.

  1. Differential expression of the cytotoxic and hemolytic activities of the ApxIIA toxin from Actinobacillus pleuropneumoniae.

    Tu, A H; Hausler, C; Young, R.; Struck, D K


    The ApxIIA protein secreted from Actinobacillus pleuropneumoniae is both hemolytic and cytotoxic. However, when the cloned apxII operon is expressed in Escherichia coli, two forms of the ApxIIA protein can be recovered. Toxin which remains intracellular has hemolytic and cytotoxic activities, while toxin that is secreted is cytotoxic with little or no hemolytic activity. This indicates that the cytotoxicity of ApxIIA is independent of its hemolytic activity.

  2. Transcriptional Profiling of Hilar Nodes from Pigs after Experimental Infection with Actinobacillus Pleuropneumoniae

    Shumin Yu


    Full Text Available The gram-negative bacterium Actinobacillus pleuropneumoniae (APP is an inhabitant of the porcine upper respiratory tract and the causative agent of porcine pleuropneumonia (PP. In recent years, knowledge about the proinflammatory cytokine and chemokine gene expression that occurs in lung and lymph node of the APP-infected swine has been advanced. However, systematic gene expression profiles on hilar nodes from pigs after infection with Actinobacillus pleuropneumoniae have not yet been reported. The transcriptional responses were studied in hilar nodes (HN from swine experimentally infected with APP and the control groupusing Agilent Porcine Genechip, including 43,603 probe sets. 9,517 transcripts were identified as differentially expressed (DE at the p ≤ 0.01 level by comparing the log2 (normalized signal of the two groups named treatment group (TG and controls (CG. Eight hundred and fifteen of these DE transcripts were annotated as pig genes in the GenBank database (DB. Two hundred and seventy-two biological process categories (BP, 75 cellular components and 171 molecular functions were substantially altered in the TG compared to CG. Many BP were involved in host immune responses (i.e., signaling, signal transmission, signal transduction, response to stimulus, oxidation reduction, response to stress, immune system process, signaling pathway, immune response, cell surface receptor linked signaling pathway. Seven DE gene pathways (VEGF signaling pathway, Long-term potentiation, Ribosome, Asthma, Allograft rejection, Type I diabetes mellitus and Cardiac muscle contraction and statistically significant associations with host responses were affected. Many cytokines (including NRAS, PI3K, MAPK14, CaM, HSP27, protein phosphatase 3, catalytic subunit and alpha isoform, mediating the proliferation and migration of endothelial cells and promoting survival and vascular permeability, were activated in TG, whilst many immunomodulatory cytokines were

  3. Reação em Cadeia da Polimerase (PCR baseada no gene cpx para detecção de Actinobacillus pleuropneumoniae em suínos natural e experimentalmente infectados Polymerase Chain Reaction (PCR based on the cpx gene for detection of Actinobacillus pleuropneumoniae in natural and experimentally infected pigs

    Karina Koerich de Souza


    Full Text Available A pleuropneumonia suína é uma das mais importantes doenças respiratórias dos suínos, estando presente em todos os países produtores. Para o controle e o monitoramento da pleuropneumonia, é necessário o desenvolvimento de métodos rápidos e acurados de diagnóstico. Com o objetivo de validar a técnica da PCR, baseada no gene cpx de Actinobacillus pleuropneumoniae, em suínos sabidamente positivos, primeiramente foi realizada inoculação experimental com amostras de A. pleuropneumoniae sorotipo 5B e coletadas amostras por meio de suabe de tonsila, biópsia de tonsila e sangue para realização da técnica de PCR, isolamento bacteriológico e teste de ELISA, respectivamente. Posteriormente, estas técnicas foram aplicadas em suínos naturalmente infectados, em três rebanhos com diferentes situações sanitárias quanto à apresentação clínica da doença. De cada rebanho, foram analisados cinco grupos de suínos com idades diferentes, sendo coletado de cada animal biópsia de tonsila para isolamento bacteriológico e PCR e sangue para determinação do perfil sorológico. Os resultados obtidos na inoculação experimental confirmaram que, mesmo com o estabelecimento da infecção comprovada pelo isolamento bacteriológico, após o período de 45 dias, não foi possível detectar o agente pela técnica de PCR. Em animais naturalmente infectados, a técnica de PCR apresentou maior sensibilidade quando comparado com o isolamento. A associação entre PCR e ELISA demonstrou ser uma boa alternativa para definir a situação sanitária do rebanho quanto à infecção por A. pleuropneumoniae.Swine pleuropneumonia is one of the most important pig respiratory diseases and has been found in all producer countries. For control and monitoring of pleuropneumonia, it is necessary the development of fast and specific methods of diagnosis. To validate PCR based on the cpx gene of Actinobacillus pleuropneumoniae in positive pigs, an experimental

  4. Conservation and antigenic cross-reactivity of the transferrin-binding proteins of Haemophilus influenzae, Actinobacillus pleuropneumoniae and Neisseria meningitidis.

    Holland, J; Parsons, T R; Hasan, A A; Cook, S M; Stevenson, P; Griffiths, E; Williams, P


    Haemophilus influenzae acquires iron from the iron-transporting glycoprotein transferrin via a receptor-mediated process. This involves two outer-membrane transferrin-binding proteins (Tbps) termed Tbp1 and Tbp2 which show considerable preference for the human form of transferrin. Since the Tbps are attracting considerable attention as potential vaccine components, we used transferrin affinity chromatography to examine their conservation amongst 28 H. influenzae type b strains belonging to different outer-membrane-protein subtypes as well as six non-typable strains. Whole cells of all type b and non-typable strains examined bound human transferrin; whilst most strains possessed a Tbp1 of approximately 105 kDa, the molecular mass of Tbp2 varied from 79 to 94 kDa. Antisera raised against affinity-purified native H. influenzae Tbp1/Tbp2 receptor complex cross-reacted on Western blots with the respective Tbps of all the Haemophilus strains examined. When used to probe Neisseria meningitidis Tbps, sera from each of four mice immunized with the Haemophilus Tbp1/2 complex recognized the 68 kDa Tbp2 of N. meningitidis strain B16B6 but not the 78 kDa Tbp2 of N. meningitidis strain 70942. Serum from one mouse also reacted weakly with Tbp1 of strain B16B6. Apart from a weak reaction with the Tbp2 of a serotype 5 strain, this mouse antiserum failed to recognize the Tbps of the porcine pathogen A. pleuropneumoniae. However, a monospecific polyclonal antiserum raised against the denatured Tbp2 of Neisseria meningitidis B16B6 recognized the Tbps of all Haemophilus and Actinobacillus strains examined. Since H. influenzae forms part of the natural flora of the upper respiratory tract, human sera were screened for the presence of antibodies to the Tbps. Sera from healthy adults contained antibodies which recognized both Tbp1 and Tbp2 from H. influenzae but not N. meningitidis. Convalescent sera from meningococcal meningitis patients contained antibodies which, on Western blots

  5. A novel Respiratory Health Score (RHS supports a role of acute lung damage and pig breed in the course of an Actinobacillus pleuropneumoniae infection

    Gerlach Gerald F


    Full Text Available Abstract Background Bacterial lung infections are a major cause of economic losses in the pig industry; they are responsible for approximately 50% of the antibiotics used in pigs and, therefore, also present an increasing concern to consumer protection agencies. In response to this changing market we investigated the feasibility of an old approach aimed at the breeding selection of more resistant pigs. As a first step in this direction we applied a new respiratory health score system to study the susceptibility of four different pig breeding lines (German Landrace, Piétrain, Hampshire, Large White towards the respiratory tract pathogen Actinobacillus (A. pleuropneumoniae. Results A controlled experimental aerosol infection with an A. pleuropneumoniae serotype 7 isolate was performed using 106 weaning pigs of defined breeding lines from the breeds German Landrace, Piétrain, Hamphire, and Large White. Pigs were clinically assessed on days 4 and 20 post infection following a novel scoring system, the Respiratory Health Score (RHS, which combines clinical, sonographic and radiographic examination results. The ranking on day 4 was significantly correlated with the ranking based on the pathomorphological Lung Lesion Score (LLS; Spearman Rank Correlation Coefficient of 0.86 [p Conclusion These results demonstrate that the RHS obtained from live pigs shows a highly significant correlation to the lung lesion score considered as a "gold standard". The correlation of the ranking at days 4 and 20 post infection implies that the course of disease is highly dependent on the acute lung damage. The different severity of signs among the tested pig breeding lines clearly suggests a genetic difference in the susceptibility of pigs to A. pleuropneumoniae infection.

  6. Microarray-based comparative genomic profiling of reference strains and selected Canadian field isolates of Actinobacillus pleuropneumoniae

    MacInnes Janet I


    Full Text Available Abstract Background Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is a highly contagious respiratory pathogen that causes severe losses to the swine industry worldwide. Current commercially-available vaccines are of limited value because they do not induce cross-serovar immunity and do not prevent development of the carrier state. Microarray-based comparative genomic hybridizations (M-CGH were used to estimate whole genomic diversity of representative Actinobacillus pleuropneumoniae strains. Our goal was to identify conserved genes, especially those predicted to encode outer membrane proteins and lipoproteins because of their potential for the development of more effective vaccines. Results Using hierarchical clustering, our M-CGH results showed that the majority of the genes in the genome of the serovar 5 A. pleuropneumoniae L20 strain were conserved in the reference strains of all 15 serovars and in representative field isolates. Fifty-eight conserved genes predicted to encode for outer membrane proteins or lipoproteins were identified. As well, there were several clusters of diverged or absent genes including those associated with capsule biosynthesis, toxin production as well as genes typically associated with mobile elements. Conclusion Although A. pleuropneumoniae strains are essentially clonal, M-CGH analysis of the reference strains of the fifteen serovars and representative field isolates revealed several classes of genes that were divergent or absent. Not surprisingly, these included genes associated with capsule biosynthesis as the capsule is associated with sero-specificity. Several of the conserved genes were identified as candidates for vaccine development, and we conclude that M-CGH is a valuable tool for reverse vaccinology.

  7. Experimental Actinobacillus pleuropneumoniae challenge in swine: Comparison of computed tomographic and radiographic findings during disease

    Brauer Carsten


    Full Text Available Abstract Background In pigs, diseases of the respiratory tract like pleuropneumonia due to Actinobacillus pleuropneumoniae (App infection have led to high economic losses for decades. Further research on disease pathogenesis, pathogen-host-interactions and new prophylactic and therapeutic approaches are needed. In most studies, a large number of experimental animals are required to assess lung alterations at different stages of the disease. In order to reduce the required number of animals but nevertheless gather information on the nature and extent of lung alterations in living pigs, a computed tomographic scoring system for quantifying gross pathological findings was developed. In this study, five healthy pigs served as control animals while 24 pigs were infected with App, the causative agent of pleuropneumonia in pigs, in an established model for respiratory tract disease. Results Computed tomographic (CT findings during the course of App challenge were verified by radiological imaging, clinical, serological, gross pathology and histological examinations. Findings from clinical examinations and both CT and radiological imaging, were recorded on day 7 and day 21 after challenge. Clinical signs after experimental App challenge were indicative of acute to chronic disease. Lung CT findings of infected pigs comprised ground-glass opacities and consolidation. On day 7 and 21 the clinical scores significantly correlated with the scores of both imaging techniques. At day 21, significant correlations were found between clinical scores, CT scores and lung lesion scores. In 19 out of 22 challenged pigs the determined disease grades (not affected, slightly affected, moderately affected, severely affected from CT and gross pathological examination were in accordance. Disease classification by radiography and gross pathology agreed in 11 out of 24 pigs. Conclusions High-resolution, high-contrast CT examination with no overlapping of organs is superior to

  8. Actinobacillus pleuropneumoniae osteomyelitis in pigs demonstrated by fluorescent in situ hybridization

    Jensen, Tim Kåre; Boye, Mette; Hagedorn-Olsen, T.;


    of A. pleuropneumoniae in formalin-fixed tissue was performed to verify the association of A. pleuropneumoniae with the bone and joint lesions. By in situ hybridization A. pleuropneumoniae was demonstrated as multiple microcolonies or single cells dispersed in focal fibrinonecrotizing pleuropneumonia...

  9. New insights on the treatment of respiratory diseases caused by actinobacillus pleuropneumoniae and Haemophilus parasuis in pigs with marbofloxacin

    Vilalta Sans, Carles


    La marbofloxacina (MB) és una fluoroquinolona de tercera generació àmpliament usada en diferents espècies per tractar sobretot infeccions respiratòries. Aquest antibiòtic posseeix un ampli espectre d’activitat que inclou dos dels principals patògens associats al complexe respiratori porcí (CRP), Actinobacillus pleuropneumoniae (APP) i Haemophilus parasuis (HP). APP és l’agent etiològic de la pleuropneumònia porcina i pot romandre a les tonsil·les dels porcs sense mostrar cap mena de símptoma ...

  10. New insights on the treatment of respiratory diseases caused by actinobacillus pleuropneumoniae and Haemophilus parasuis in pigs with marbofloxacin

    Vilalta Sans, Carles; Cristòfol Adell, Carles


    La marbofloxacina (MB) és una fluoroquinolona de tercera generació àmpliament usada en diferents espècies per tractar sobretot infeccions respiratòries. Aquest antibiòtic posseeix un ampli espectre d'activitat que inclou dos dels principals patògens associats al complexe respiratori porcí (CRP), Actinobacillus pleuropneumoniae (APP) i Haemophilus parasuis (HP). APP és l'agent etiològic de la pleuropneumònia porcina i pot romandre a les tonsil·les dels porcs sense mostrar cap mena de símptoma ...

  11. Real-time quantitative reverse transcription-PCR analysis of expression stability of Actinobacillus pleuropneumoniae housekeeping genes during in vitro growth under iron-depleted conditions

    Nielsen, K. K.; Boye, Mette


    The aims of the present investigation were to develop and test a sensitive and reproducible method for the study of gene expression in the porcine lung pathogen Actinobacillus pleuropneumoniae by real-time quantitative reverse transcription (RT)-PCR and to evaluate a number of suitable internal c...

  12. Pharmacokinetics of tulathromycin in edible tissues of healthy and experimentally infected pigs with Actinobacillus pleuropneumoniae.

    Bladek, Tomasz; Posyniak, Andrzej; Jablonski, Artur; Gajda, Anna


    The aim of this study was the comparison of the tissue pharmacokinetics of tulathromycin in healthy pigs and pigs experimentally infected with Actinobacillus pleuropneumoniae (App). Tulathromycin was given to 24 healthy and 24 infected pigs by intramuscular injection at a single dosage of 2.5 mg kg(-1) body weight (b.w.). Pigs were euthanised at each group and then samples of liver, kidney, muscle, injection site and skin with fat were taken at scheduled time points. Drug concentrations were determined by LC-MS/MS. In this study, higher values of the area under the concentration-time curves (AUC) were calculated in all tissue samples taken from infected than healthy pigs. In pigs with App the AUCs of liver, kidney, muscle, skin with fat and injection site were 1111, 1973, 235, 181 and 2931 mg kg(-1) h, while in pigs without inflammation they were 509, 1295, 151, 111 and 1587 mg kg(-1) h, respectively. Maximum drug tissue concentrations (Cmax) in infected animals were 2370, 6650, 2016, 666 and 83,870 µg kg(-1), while in healthy pigs they were 1483, 6677, 1733, 509 and 55,006 µg kg(-1), respectively. The eliminations half-times (T1/2) were respectively longer in all tissue samples taken from infected animals (from 157.3 to 187.3 h) than in healthy ones (from 138.6 to 161.2 h). The tulathromycin tissue concentrations were significantly higher (p < 0.05) in all tissue samples of the infected pigs compared with the healthy animals at 360 h (from 0.0014 to 0.0280) and at 792 h (from 0.0007 to 0.0242) after drug administration. The results suggest that the tissue pharmacokinetic properties and residue depletion of tulathromycin can be influenced by the disease state of animals. PMID:26247868

  13. Construction and immunogenicity of a ∆apxIC/ompP2 mutant of Actinobacillus pleuropneumoniae and Haemophilus parasuis

    Qiong Liu


    Full Text Available The apxIC genes of the Actinobacillus pleuropneumoniae serovar 5 (SC-1, encoding the ApxIactivating proteins, was deleted by a method involving sucrose counter-selection. In this study, a mutant strain of A. pleuropneumoniae (SC-1 was constructed and named DapxIC/ ompP2. The mutant strain contained foreign DNA in the deletion site of ompP2 gene of Haemophilus parasuis. It showed no haemolytic activity and lower virulence of cytotoxicity in mice compared with the parent strain, and its safety and immunogenicity were also evaluated in mice. The LD50 data shown that the mutant strain was attenuated 30-fold, compared with the parent strain (LD50 of the mutant strain and parent strain in mice were determined to be 1.0 × 107 CFU and 3.5 × 105 CFU respectively. The mutant strain that was attenuated could secrete inactivated ApxIA RTX toxins with complete antigenicity and could be used as a candidate live vaccine strain against infections of A. pleuropneumoniae and H. parasuis.

  14. Estudio del comportamiento serológico de Actinobacillus pleuropneumoniae (App en planteles porcinos comerciales de la zona central de Chile Serological behaviour study of Actinobacillus pleuropneumoniae (App in commercial swine herds from the central region of Chile

    D Muñoz


    Full Text Available En Chile se ha realizado sólo un estudio en Actinobacillus pleuropneumoniae (App. Este trabajo pretende determinar la duración de la inmunidad materna, la edad de seroconversión y la prevalencia aparente y verdadera en 7 planteles de cerdos comerciales. Se obtuvieron 60 muestras por plantel, divididas en 10 muestras de suero, de animales de 4, 6, 10, 14,18 y 21 semanas de edad, y analizadas a través de un kit ELISA® comercial. De las 420 muestras se detectaron 134 positivas, de las cuales 112 correspondían a cerdos menores de 10 semanas y sólo 22 provenían de animales mayores de 10 semanas, que seroconvirtieron probablemente debido a una infección de campo. La caída de la inmunidad materna fue alrededor de la 10ª semana de edad. En cuanto a la seroconversión, se observó que a partir de la 18* semana comenzaron a aparecer los animales con anticuerpos circulantes propios. Dos de los siete planteles no seroconvirtieron. Además, dos presentaron una seroconversión igual o superior al 50% a las 18 semanas. La seroprevalencia aparente de App fue de 10,48%, mientras que prevalencia verdadera, mediante dos métodos estadísticos, fue de 9,6% (IC: 7,6% y 11,7% y 10,67% respectivamente. En este trabajo se encontró que la prevalencia es similar a la observada en EE.UU., debido presumiblemente al sistema de producción y a los serotipos que están presentes en ambos países. Por otro lado, si bien la mayoría de los planteles seroconvierten luego de la caída de la inmunidad materna, se observaron diferentes patrones serológicos entre ellos.In Chile, there was only one existing study on App. This study was designed to determine the maternal immunity duration, the age of seroconversion and the apparent and true prevalence in animals from 7 swine commercial herds. 60 samples were taken per herd and divided into 10 serum samples from animals of 4, 6,10,14,18and21 weeks of age, which were analyzed by ELISA®. Out of the 420 samples, 134 were

  15. Molecular characterisation of the early response in pigs to experimental infection with Actinobacillus pleuropneumoniae using cDNA microarrays

    Bendixen Christian


    Full Text Available Abstract Background The bacterium Actinobacillus pleuropneumoniae is responsible for porcine pleuropneumonia, a widespread, highly contagious and often fatal respiratory disease of pigs. The general porcine innate immune response after A. pleuropneumoniae infection is still not clarified. The objective of this study was hence to characterise the transcriptional response, measured by using cDNA microarrays, in pigs 24 hours after experimental inoculation with A. pleuropneumoniae. Methods Microarray analyses were conducted to reveal genes being differentially expressed in inflamed versus non-inflamed lung tissue sampled from inoculated animals as well as in liver and tracheobronchial lymph node tissue sampled from three inoculated animals versus two non-inoculated animals. The lung samples were studied using a porcine cDNA microarray with 5375 unique PCR products while liver tissue and tracheobronchial lymph node tissue were hybridised to an expanded version of the porcine microarray with 26879 unique PCR products. Results A total of 357 genes differed significantly in expression between infected and non-infected lung tissue, 713 genes differed in expression in liver tissue from infected versus non-infected animals and 130 genes differed in expression in tracheobronchial lymph node tissue from infected versus non-infected animals. Among these genes, several have previously been described to be part of a general host response to infections encoding immune response related proteins. In inflamed lung tissue, genes encoding immune activating proteins and other pro-inflammatory mediators of the innate immune response were found to be up-regulated. Genes encoding different acute phase reactants were found to be differentially expressed in the liver. Conclusion The obtained results are largely in accordance with previous studies of the mammalian immune response. Furthermore, a number of differentially expressed genes have not previously been associated

  16. Impact of Actinobacillus pleuropneumoniae biofilm mode of growth on the lipid A structures and stimulation of immune cells.

    Hathroubi, Skander; Beaudry, Francis; Provost, Chantale; Martelet, Léa; Segura, Mariela; Gagnon, Carl A; Jacques, Mario


    Actinobacillus pleuropneumoniae (APP), the etiologic agent of porcine pleuropneumonia, forms biofilms on biotic and abiotic surfaces. APP biofilms confers resistance to antibiotics. To our knowledge, no studies have examined the role of APP biofilm in immune evasion and infection persistence. This study was undertaken to (i) investigate biofilm-associated LPS modifications occurring during the switch to biofilm mode of growth; and (ii) characterize pro-inflammatory cytokines expression in porcine pulmonary alveolar macrophages (PAMs) and proliferation in porcine PBMCs challenged with planktonic or biofilm APP cells. Extracted lipid A samples from biofilm and planktonic cultures were analyzed by HPLC high-resolution, accurate mass spectrometry. Biofilm cells displayed significant changes in lipid A profiles when compared with their planktonic counterparts. Furthermore, in vitro experiments were conducted to examine the inflammatory response of PAMs exposed to UV-inactivated APP grown in biofilm or in suspension. Relative mRNA expression of pro-inflammatory genes IL1, IL6, IL8 and MCP1 decreased in PAMs when exposed to biofilm cells compared to planktonic cells. Additionally, the biofilm state reduced PBMCs proliferation. Taken together, APP biofilm cells show a weaker ability to stimulate innate immune cells, which could be due, in part, to lipid A structure modifications. PMID:27226465

  17. Selective media for the isolation of Actinobacillus pleuropneumoniae from the pig

    Vidić Branka M.


    Full Text Available Selective media were developed and evaluated for isolation of A. pleuropneumoniae from pig tonsils. Samples were obtained from four pig herds with a clinical history of pleuropneumonia. For isolation of A. pleuropneumoniae 93 pig tonsils were collected at slaughter. Each sample was streaked on to four different selective media (modified PPLO agar (mPPLO Brain-Heart agar (BH, Columbia agar (CA, Miller-Hinton chocolate agar (MHCA containing different combinations of antibiotics, NAD and nystatin. The selectivity of nutritive media is conditioned by the content of antibiotics, as well as by the type of medium used. Mean isolation rate of A. pleuropneumoniae in the investigated herds, was 17.2%. The best results were obtained using PPLO2 agar, 20.4%. The other media supplemented with the mentioned antibiotics gave satisfactory results.

  18. Identification and Detection of Actinobacillus pleuropneumoniae in Infected and Subclinically Infected Pigs by Multiplex PCR Based on the Genes ApxIVA and OmlA

    XIAO Guo-sheng; CAO San-jie; DUAN Li-li; WEN Xin-tian; MA Xiao-ping; CHEN Hua-mei


    PCRs based on different genes of Actinobacillus pleuropneumoniae have been developed for detecting and identifying A. pleuropneumoniae. Some of them could amplify positive fragments from the phylogenetically closely related species bacteria. To improve veracity and specificity of PCR, a species-specific multiplex PCR assay was developed to identify and detect A. pleuropneumoniae, based on the 3'-terminus of the species-specific apxIVA gene and the already existing species-specific primers in the omlA gene. Both 346-bp and 950-bp fragments could be simultaneously amplified from all A. pleuropneumoniae reference strains and isolates, and the species specificity of the assay was evaluated with a collection of ten strains representing eight different species bacteria including species normally found in the respiratory tracts of swine. All of these strains turned out negative in the multiplex PCR. All sequences of products of multiplex PCR randomly sampled were also correct. The sensitivity of the multiplex PCR was determined to be 10 pg ofA. pleuropneumoniae DNA. The multiplex PCR and bacterial isolation were compared to determine their sensitivities by using experimentally infected pigs and clinical disease pigs. The multiplex PCR was more sensitive than bacterial isolation. The multiplex PCR was also evaluated on mixed bacterial cultures from clinical healthy pigs. 26/100 (26%) of the subclinically infected pigs were detected from clinical healthy pigs. The results indicate that the multiplex PCR assay is a sensitive, highly specific,and effective diagnostic tool for identification and detection of A. pleuropneumoniae.

  19. Economic impacts of reduced pork production associated with the diagnosis of Actinobacillus pleuropneumoniae on grower/finisher swine operations in the United States.

    Losinger, Willard C


    An examination of the economic impacts of the diagnosis of Actinobacillus pleuropneumoniae on grower/finisher swine operations indicated that reduced pork production, associated with the diagnosis of A. pleuropneumoniae on the operation, diminished consumer surplus by $53+/-52 million, and resulted in a total loss of $32+/-30 million to the US economy in 1995. Most of the economic surplus lost by consumers was transferred to producers, whose economic surplus increased by $21+/-25 million (which was not significantly different from zero). Uncertainty analysis showed that an estimate of the decline in production associated with the diagnosis of A. pleuropneumoniae accounted for most of the uncertainty of the change in consumer surplus and of the total loss to the economy. The estimate of the price elasticity of demand for pork also contributed towards a lot of the uncertainty in the estimated change in producer surplus. PMID:15820115

  20. Evaluation and application of ribotyping for epidemiological studies of Actinobacillus pleuropneumoniae in Denmark

    Fussing, V.; Barfod, Kristen; Nielsen, R.;


    discriminatory power was between 0.85-0.89. The relatively low discriminatory power was caused by four predominant types, containing 61% of the isolates. The typing system was applied in studies of routes of infection of specific pathogen-free (SPF) pig herds and included 112 strains of A. pleuropneumoniae...

  1. Activity of antibodies against Salmonella dublin, Toxoplasma gondii, or Actinobacillus pleuropneumoniae in sera after treatment with electron beam irradiation or binary ethylenimine

    Kyvsgaard, N.C.; Lind, Peter; Preuss, T.; Kamstrup, Søren; Lei, J.C.; Bogh, H.O.; Nansen, P.


    used as an estimate for the relative posttreatment activity. For a Toxoplasma gondii indirect enzyme-linked immunosorbent assay (ELISA) and agglutination assay as well as for a Salmonella dublin indirect ELISA, the posttreatment activity was more than 89% of the pretreatment activity when the samples...... were irradiated in the frozen state on dry ice but only 35% of their activity when they were irradiated in the liquid state at 0 degrees C. The patterns seen in an S. dublin blocking ELISA and an Actinobacillus pleuropneumoniae complement fixation assay differed in that samples with a low level of...

  2. The porcine acute phase response to infection with Actinobacillus pleuropneumoniae. Haptoglobin, C-reactive protein, major acute phase protein and serum amyloid a protein are sensitive indicators of infection

    Heegaard, Peter M. H.; Klausen, Joan; Nielsen, J.P.;


    In an experimental infection model mimicking acute Actinobacillus pleuropneumoniae (Ap) infection in swine (Sus scrofa) by aerosol inoculation, the development of a number of typical clinical signs was accompanied by a prototypic acute phase reaction encompassing fever and an acute phase protein ...

  3. An Actinobacillus pleuropneumoniae PCR typing system based on the apx and omlA genes - evaluation of isolates from lungs and tonsils of pigs

    Gram, T.; Ahrens, Peter; Andreasen, Morten;


    . The PCR typing system was tested on 102 field strains of A. pleuropneumoniae isolated from lungs of diseased pigs. The serotyping results of the investigated field strains were in agreement with the apr and omlA gene patterns found in the reference strains of the bacteria, with the exception of the...... omlA gene of five strains of serotype 8. To examine the apx and omlA gene pattern of tonsil isolates, the PCR typing system was tested on a total of 280 A. pleuropneumoniae field strains isolated from tonsils of pigs. Agreement between serotyping and DNA typing was found in 96% of the isolates using...... the apx gene patterns and in 89% of the isolates using the omlA gene. The same serotype specific apx/omlA gene pattern was thus found in the majority of the tonsil isolates and in isolates from diseased lungs. Most of the differences in the omlA gene were found in 18 tonsil isolates of serotype 12...

  4. Expresión recombinante en E. Coli de antígenos de Actinobacillus Pleuropneumoniae para vacunación y diagnóstico

    Medrano Muñoz, Andrés


    Actinobacillus pleuropneumoniae es una bacteria gramnegativa que provoca la pleuroneumonía porcina. En este trabajo se ha procedido a la producción y purificación, mediante técnicas de biología molecular, de antígenos proteicos de esta bacteria y a su uso en la formulación de una vacuna por subunidades y de un ELISA para diagnóstico. Los cuatro antígenos escogidos fueron dos proteínas de membrana externa (Tbp1 y Tbp2) y dos exotoxinas (ApxI y ApxIII). La primera necesidad consistía en localiz...

  5. Comparative activities of selected fluoroquinolones against dynamic populations of Actinobacillus pleuropneumoniae in an in vitro model of time-kill continuous culture experiment.

    Damte, Dereje; Lee, Seung-Jin; Yohannes, Sileshi B; Hossain, Md Akil; Suh, Joo-Won; Park, Seung-Chun


    The aim of the current study was to demonstrate and compare the impact of different pharmacokinetics of marbofloxacin, enrofloxacin and difloxacin on their antimicrobial effects, their killing and re-growth kinetics, and the population dynamics of Actinobacillus pleuropneumoniae clinical isolates in an in vitro dynamic model. Selected clinical isolates of A. pleuropneumoniae and three fluoroquinolones at a range of simulated AUC(24)/MIC ratios of multiple doses were investigated. At the same simulated AUC(24)/MIC ratios of the three fluoroquinolones, the killing re-growth profile and I(E) values (intensity of the antimicrobial effect) revealed strain- and fluoroquinolone-specific effects. For example, a 31% lower I(E) of difloxacin was observed in AppK5 (biofilm-former) than in AppK2 (biofilm-non-former) at the same AUC(24)/MIC ratio of 120 h. In addition, losses in A. pleuropneumoniae susceptibility of both strains by the three fluoroquinolones were observed. AUC(24)/MPC ratios of 20.89 and 39.81 for marbofloxacin, 17.32 and 19.49 for enrofloxacin and 31.62 and 60.25 for difloxacin were estimated to be protective against the selection of AppK2 and AppK5 strain mutants, respectively. Integration of these in vitro data with published pharmacokinetics revealed the inadequacy of the conventional clinical doses of the three drugs to attain the above protective values for minimum biofilm eradication concentration (MBEC) and concentration to prevent growth of 90% of the mutant subpopulation (MPC(90)). In conclusion, the results suggest optimising doses could suffice for resistant mutants control, while for biofilm-forming strains combination with biofilm-disrupting agents to reduce the MBEC to achieve AUC/MBEC ratios within the possible dosing regimens is desired. PMID:24139884

  6. Antioxidant effect of minocycline in gingival epithelium induced by Actinobacillus actinomycetemcomitans serotype B toxin

    Ernie Maduratna Setiawati


    Full Text Available Background: Actinobacillus actinomycetemcomitans (Aa serotype B has been associated with aggressive periodontitis. Gingival epithelial cell is exquisitely sensitive to the toxin and may lead to the epithel protective barrier disruption. Experimental models show that minocycline is not related to it’s antimicrobial effect and protection against neuron cell apoptosis of a number experimental models of brain injury and Parkinson’s disease. Purpose: This study, examined antioxidant effect of minocycline to inhibit apoptosis of gingival epithelium induced crude toxin bacteria Aa serotype B in mice. Methods: Thirty adult mice strain Swiss Webster (balb C were divided randomly into three groups: control group (group A, toxin group (group B and toxin and minocycline group (group C. The mice were taken at 24 hours after application, and then the tissue sections of gingival epithelium were stained with tunnel assay and immunohistochemistry. Result: Treatment with these toxin induced apoptosis of gingival epithelium and was associated with DNA fragmentation and reduced gluthatione (GSH. Minocycline 100 nM significantly increased GSH and reduced apoptosis (p < 0.05. Minocycline provides antioxidant effect against citotoxicity of bacteria Aa serotipe B. Conclusion: Nanomolar concentration of minocycline potential as new therapeutic agent to prevent progressivity of aggressiveness of periodontitis.


    Jakobsen, Marianne; Nielsen, Jens

    In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A. pleuropn......In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A...


    Jakobsen, Marianne; Nielsen, Jens


    In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A. pleuropn......In order to isolate ActinobacillIus pleuropneumoniae from mixed bacterial flora a selective and indicative medium was developed. The optimal concentrations of antibiotics were determined for selective chocolate agar (S-TSA) and selective blood agar (S-MBA) using a set of 25 strains of A...

  9. Transcriptional Portrait of Actinobacillus pleuropneumoniae during Acute Disease - Potential Strategies for Survival and Persistence in the Host

    Schou, Kirstine Klitgaard; Rundsten, Carsten Friis; Jensen, Tim Kåre; Angen, Øystein; Boye, Mette


    results suggested that A. pleuropneumoniae is using various strategies to increase its fitness, such as applying Na+ pumps as an alternative way of gaining energy. Furthermore, the transcriptional data provided potential clues as to how A. pleuropneumoniae is able to circumvent host immune factors and...... survive within the hostile environment of host macrophages. This persistence within macrophages may be related to urease activity, mobilization of various stress responses and active evasion of the host defenses by cell surface sialylation. Conclusions/Significance The data presented here highlight the...

  10. Enriched Housing Reduces Disease Susceptibility to Co-Infection with Porcine Reproductive and Respiratory Virus (PRRSV) and Actinobacillus pleuropneumoniae (A. pleuropneumoniae) in Young Pigs.

    van Dixhoorn, Ingrid D E; Reimert, Inonge; Middelkoop, Jenny; Bolhuis, J Elizabeth; Wisselink, Henk J; Groot Koerkamp, Peter W G; Kemp, Bas; Stockhofe-Zurwieden, Norbert


    Until today, anti-microbial drugs have been the therapy of choice to combat bacterial diseases. Resistance against antibiotics is of growing concern in man and animals. Stress, caused by demanding environmental conditions, can reduce immune protection in the host, influencing the onset and outcome of infectious diseases. Therefore psychoneuro-immunological intervention may prove to be a successful approach to diminish the impact of diseases and antibiotics use. This study was designed to investigate the effect of social and environmental enrichment on the impact of disease, referred to as "disease susceptibility", in pigs using a co-infection model of PRRSV and A. pleuropneumoniae. Twenty-eight pigs were raised in four pens under barren conditions and twenty-eight other pigs were raised in four pens under enriched conditions. In the enriched pens a combination of established social and environmental enrichment factors were introduced. Two pens of the barren (BH) and two pens of the enriched housed (EH) pigs were infected with PRRSV followed by A. pleuropneumoniae, the other two pens in each housing treatment served as control groups. We tested if differences in disease susceptibility in terms of pathological and clinical outcome were related to the different housing regimes and if this was reflected in differences in behavioural and immunological states of the animals. Enriched housed pigs showed a faster clearance of viral PRRSV RNA in blood serum (p = 0.014) and histologically 2.8 fold less interstitial pneumonia signs in the lungs (p = 0.014). More barren housed than enriched housed pigs developed lesions in the lungs (OR = 19.2, p = 0.048) and the lesions in the barren housed pigs showed a higher total pathologic tissue damage score (pdisease susceptibility to co-infection of PRRSV and A. pleuropneumoniae in pigs. Enrichment positively influences behavioural state, immunological response and clinical outcome in pigs. PMID:27606818

  11. The Actinobacillus pleuropneumoniae HMW1C-like glycosyltransferase mediates N-linked glycosylation of the Haemophilus influenzae HMW1 adhesin.

    Kyoung-Jae Choi

    Full Text Available The Haemophilus influenzae HMW1 adhesin is an important virulence exoprotein that is secreted via the two-partner secretion pathway and is glycosylated at multiple asparagine residues in consensus N-linked sequons. Unlike the heavily branched glycans found in eukaryotic N-linked glycoproteins, the modifying glycan structures in HMW1 are mono-hexoses or di-hexoses. Recent work demonstrated that the H. influenzae HMW1C protein is the glycosyltransferase responsible for transferring glucose and galactose to the acceptor sites of HMW1. An Actinobacillus pleuropneumoniae protein designated ApHMW1C shares high-level homology with HMW1C and has been assigned to the GT41 family, which otherwise contains only O-glycosyltransferases. In this study, we demonstrated that ApHMW1C has N-glycosyltransferase activity and is able to transfer glucose and galactose to known asparagine sites in HMW1. In addition, we found that ApHMW1C is able to complement a deficiency of HMW1C and mediate HMW1 glycosylation and adhesive activity in whole bacteria. Initial structure-function studies suggested that ApHMW1C consists of two domains, including a 15-kDa N-terminal domain and a 55-kDa C-terminal domain harboring glycosyltransferase activity. These findings suggest a new subfamily of HMW1C-like glycosyltransferases distinct from other GT41 family O-glycosyltransferases.

  12. Differences in Purinergic Amplification of Osmotic Cell Lysis by the Pore-Forming RTX Toxins Bordetella pertussis CyaA and Actinobacillus pleuropneumoniae ApxIA: the Role of Pore Size

    Mašín, Jiří; Fišer, Radovan; Linhartová, Irena; Osička, Radim; Bumba, Ladislav; Hewlett, E. L.; Benz, R.; Šebo, Peter


    Roč. 81, č. 12 (2013), s. 4571-4582. ISSN 0019-9567 R&D Projects: GA ČR GAP302/12/0460; GA ČR(CZ) GAP302/11/0580; GA ČR(CZ) GAP207/11/0717; GA AV ČR IAA500200914; GA ČR GA13-14547S Institutional support: RVO:61388971 Keywords : Bordetella pertussis * Actinobacillus pleuropneumoniae * E-coli Subject RIV: EE - Microbiology, Virology Impact factor: 4.156, year: 2013

  13. Delineation of the genus Actinobacillus by comparison of partial infB sequences

    Nørskov-Lauritsen, Niels; Christensen, H; Okkels, H.; Kilian, Mogens; Bruun, B


    position of Actinobacillus capsulatus was unresolved; this species is more remotely related to A. lignieresii. The two species A. lignieresii and A. pleuropneumoniae could not be clearly separated by infB sequence analysis. The phylogeny of the genus Actinobacillus based on infB analysis was essentially......, Actinobacillus pleuropneumoniae, Actinobacillus equuli, Actinobacillus suis, Actinobacillus ureae, Actinobacillus arthritidis, Actinobacillus hominis and two unnamed genomospecies showed more than 85 % similarity to the type strain of the type species of the genus, Actinobacillus lignieresii. The taxonomic...... congruent with relationships inferred from 16S rRNA sequence comparisons and DNA hybridization studies. Discrepancies were encountered with single strains or taxa at the periphery of the genus. Greater intraspecies variation was observed with infB sequences than with 16S rRNA gene sequences, with notable...

  14. Hepatic gene expression changes in pigs experimentally infected with the lung pathogen Actinobacillus pleuropneumoniae as analysed with an innate immunity focused microarray

    Skovgaard, Kerstin; Mortensen, Shila; Boye, Mette;


    differentially expressed. A large group of these genes encoded proteins involved in the acute phase response, including serum amyloid A, C-reactive protein, fibrinogen, haptoglobin and tumor necrosis factor-a the expression of which were all found to be up-regulated and glutathione S-transferase, transthyretin...... initiating and orchestrating the innate immune response to A. pleuropneumoniae infection. Keywords: acute phase protein, hepatic transcriptional response, innate defence, gene expression, pig......, mannan-binding lectin A, surfactant protein D, and surfactant protein A1 were down-regulated in the liver of infected animals. Down-regulation of a1-acid glycoprotein during infection has not been described previously in any species. These results confirm that the liver plays an important role in...

  15. Incidence of Reinfections with Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae in Pig Farms Located in Respiratory-Disease-Free Regions of Switzerland – Identification and Quantification of Risk Factors

    Scheidegger R


    Full Text Available The objective of the study was to identify risk factors for reintroduction of Actinobacillus pleuopneumoniae and Mycoplasma hyopneumoniae (enzootic pneumonia onto pig farms in areas in Switzerland that were involved in an eradication programme from 1996 to 1999 and to assess the role of dealers in relation to these reinfections. The study was based on the comparison of pig farms that were reinfected in the year 2000 (cases and pig farms that remained uninfected in the same area (controls. Additionally, data were collected from Swiss pig dealers and transport companies. Out of a total of 3983 farms, 107 farms were reinfected in the year 2000. The incidences were 0.1% for Actinobacillus pleuopneumoniae and 2.6% for Mycoplasma hyopneumoniae (enzootic pneumonia. Compared to reinfection rates prior to the eradication programme, this is a considerable reduction. Statistically significant risk factors for the reinfection were 'finishing farm', 'large mixed breeding-finishing farm', 'reinfected neighbour' and 'parking site for pig transport vehicles close to the farm'. Pig farmers that purchased pigs from only one supplier per batch had a lower risk of reintroducing infection (protective factor. As long as infected and uninfected regions co-exist in Switzerland, direct and indirect contact between farms, pig herds and slaughter sites via transport vehicles are a major pathway of disease spread. Risk management measures linked to these contacts are therefore of key importance. The survey of dealers indicated various areas for improvement such as strategic planning of pick-up routes or cleaning and disinfecting of trucks.

  16. Serotype Identification and Medicine Sensitivity Test of Actinobcillus pleuropneumoniae%猪传染性胸膜肺炎放线杆菌的血清型鉴定与药敏试验

    蔡丙严; 周建强; 陈长春; 魏冬霞; 张步彩


    TZA1,TZA2 and TZA3 strains were obtained by bacteria separate and culture from a suspected infectious pleurop-neumoniae ill pigs in Taizhou, Jiangsu province. After morphology check, cAMP test, satellite growth phenomenon, biochemical test and serotype identification, the isolated strains were serotyped, TZA1 and TZA2 as serotype 1, TZA3 as serotype 3. And the drug sensitivity test showed that three isolated strains were high-sensitive to some antibacterial agents such as en-rofloxacin, azithromycin and ceftizoxime.%从江苏泰州地区疑似猪传染性胸膜肺炎发病猪采集病料,经细菌分离培养得到了3株分离株TZA1、TZA2和TZA3株.经形态学检查、CAMP试验、生化试验和血清型鉴定,确定TZA1株和TZA2株为血清1型,TZA3株为血清3型.药敏试验显示,3株分离菌株对恩诺沙星、阿齐霉素、头孢唑肟高度敏感.

  17. Comparison of Common Antibiotic Therapies for Haemophilus Pleuropneumonia in Pigs

    Willson, Philip J.; Osborne, A. Dudley


    Three experiments were done to evaluate some antibiotic therapies that are used commonly to treat pigs infected with Haemophilus pleuropneumoniae. Haemophilus-free piglets, 12 weeks of age, were challenged in a chamber with an aerosol of H. pleuropneumoniae serotype 1 and were medicated with antibiotics at various times before or after challenge. Antibiotic formulations which are commonly used to treat pneumonia in swine were used. They were chloramphenicol, penicillin, and a long-acting form...

  18. Comparison of six typing methods for Actinobacillus actinomycetemcomitans.

    van Steenbergen, T J; Bosch-Tijhof, C J; van Winkelhoff, A J; Gmür, R; de Graaff, J


    Actinobacillus actinomycetemcomitans is an important pathogen in the etiology of severe periodontitis. For epidemiological studies on the prevalence of certain pathogenic clones and transmission of this bacterium, adequate typing methods are necessary. The purpose of this study was to compare six different typing methods for A. actinomycetemcomitans. Five reference strains and 27 fresh clinical isolates from periodontitis patients were used. Serotyping showed 12 serotype a strains, 13 type b ...

  19. Expression of coding (mRNA) and non-coding (microRNA) RNA in lung tissue and blood isolated from pigs suffering from bacterial pleuropneumonia

    Skovgaard, Kerstin; Schou, Kirstine Klitgaard; Wendt, Karin Tarp;


    infected with Actinobacillus pleuropneumoniae (AP). Expression differences of mRNA and microRNA were quantified at different time points (6h, 12h, 24h, 48h PI) using reverse transcription quantitative real-time PCR (Rotor-Gene and Fluidigm). Expression profiles of miRNA in blood of seven animals were...

  20. Construction of the Gene Deleted Attenuated Mutant Strain apxIIC -/kanr + of Actinobacillus pleuropneumoniae%胸膜肺炎放线杆菌apxlIC7kanr+基因缺失减毒株的构建

    李建; 曹三杰; 文心田; 黄小波; 都启晶; 张明; 余慧


    The recombinant transfer vector pBSKA was electroporated into parent strain Actinobacillus pleuropneumoniae serovar 7 (APP -7 ) strain WF83. Product of the electroporation was plated onto TSB agar containing kanamyeine (Kan). After 2 days the recombinant strains were selected. Resistance of kanamycine experiment confirmed that mutant strain can counteract kanamycine. Dependence experiment of NAD confirmed that mutant strain needed NAD in growth. Identification of PCR confirmed that complete apxIIC gene was substitute for kanamycine resistance gene and there was no presence of pBSKA. Hemolytic experiment confirmed that mutant strain had no ability of haemolysis. Cytotoxicity test confirmed that mutant strain had no cytotoxieity. Safety experiment of injected mice confirmed that eytotoxicity and haemolysis of mutant was attenuated significantly so that mutant was safe to mice. Experiment of genetic stability confirmed that kanamycine resistance of mutant was stable 30 successive generations in vitro and 10 generations in vivo. All of the above - mentioned tests indicated that the gene deleted attenuated strain was constructed successfully, which provided certain basis for further genetic live vaccine research with mutant strain.%重组转移载体pBSKA通过电转化导入亲本菌胸膜肺炎放线杆菌血清7型(APP-7)WF83株,电转化后的产物涂布于TSB/Kan平板,2d获得突变株。卡那霉素抗性实验证实突变株有卡那霉素抗性;NAD依赖性实验证实突变株依赖NAD生长;PCR鉴定证实了卡那霉素抗性基因置换了apxlIC基因,并证实突变株中无pBSKA质粒的存在;溶血活性实验证实突变株完全失去了溶血活性;细胞毒性实验证实突变株的细胞毒性完全丧失;对小鼠的安全性实验证实突变株的毒力显著减弱,突变株对小鼠是安全的;遗传稳定性实验证实,突变株在体外连续传30代和在体内传10代均不会发生卡

  1. Estimation of sensitivity, specificity and predictive values of two serologic tests for the detection of antibodies against Actinobacillus pleuropneumoniae serotype 2 in the absence of a reference test (gold standard)

    Enøe, Claes; Andersen, Søren; Sørensen, Vibeke; Willeberg, Preben

    independence was assumed to models allowing for conditional dependence, given the true disease status. No strong evidence of conditional dependence in either test sensitivity or specificity was found. Assuming independence, maximum-likelihood estimates and 95% confidence intervals of the sensitivity and...

  2. Nasal immunization with mannan-decorated mucoadhesive HPMCP microspheres containing ApxIIA toxin induces protective immunity against challenge infection with Actinobacillus pleuropneumoiae in mice.

    Li, Hui-Shan; Shin, Min-Kyoung; Singh, Bijay; Maharjan, Sushila; Park, Tae-Eun; Kang, Sang-Kee; Yoo, Han-Sang; Hong, Zhong-Shan; Cho, Chong-Su; Choi, Yun-Jaie


    The development of subunit mucosal vaccines requires an appropriate delivery system or an immune modulator such as an adjuvant to improve antigen immunogenicity. The nasal route for vaccine delivery by microparticles has attracted considerable interest, although challenges such as the rapid mucociliary clearance in the respiratory mucosa and the low immunogenicity of subunit vaccine still remain. Here, we aimed to develop mannan-decorated mucoadhesive thiolated hydroxypropylmethyl cellulose phthalate (HPMCP) microspheres (Man-THM) that contain ApxIIA subunit vaccine - an exotoxin fragment as a candidate for a subunit nasal vaccine against Actinobacillus pleuropneumoniae. For adjuvant activity, mucoadhesive thiolated HPMCP microspheres decorated with mannan could be targeted to the PRRs (pathogen recognition receptors) and mannose receptors (MR) of antigen presenting cells (APCs) in the respiratory immune system. The potential adjuvant ability of Man-THM for intranasal immunization was confirmed by in vitro and in vivo experiments. In a mechanistic study using APCs in vitro, it was found that Man-THM enhanced receptor-mediated endocytosis by stimulating the MR of APCs. In vivo, the nasal vaccination of ApxIIA-loaded Man-THM in mice resulted in higher levels of mucosal sIgA and serum IgG than mice in the ApxIIA and ApxIIA-loaded THM groups due to the specific recognition of the mannan in the Man-THM by the MRs of the APCs. Moreover, ApxIIA-containing Man-THM protected immunized mice when challenged with strains of A. pleuropneumoniae serotype 5. These results suggest that mucoadhesive Man-THM may be a promising candidate for a nasal vaccine delivery system to elicit systemic and mucosal immunity that can protect from pathogenic bacteria infection. PMID:27189136

  3. tfoX (sxy)-dependent transformation of Aggregatibacter (Actinobacillus) actinomycetemcomitans

    Bhattacharjee, Mrinal K.; Fine, Daniel H.; Figurski, David H.


    tfoX (sxy) is a regulatory gene needed to turn on competence genes. Aggregatibacter (Actinobacillus) actinomycetemcomitans has a tfoX gene that is important for transformation. We cloned this gene on an IncQ plasmid downstream of the inducible tac promoter. When this plasmid was resident in cells of A. actinomycetemcomitans and tfoX was induced, the cells became competent for transformation. Several strains of A. actinomycetemcomitans, including different serotypes, as well as rough (adherent...

  4. Construction of An Activation of Toxin 1 Gene C (△apxIC) Mutant of Actinobacillus pleuropneumoniae Gomphosised Outer Membrane Protein P2 Gene(ompP2) of Haemophilus parasuis%猪传染性胸膜肺炎放线杆菌缺失毒素1激活基因C(ApxIC)同时嵌合副猪嗜血杆菌外膜蛋白P2基因(ompP2)复合突变株的构建

    刘琼; 龚雨恒; 王国镔; 文心田; 黄小波; 曹三杰


    猪胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,APP)以及副猪嗜血杆菌(Haemophilus parasuis,Hps)是引起猪发病的2种病菌.本研究以APP血清5型分离株(SW1)为基础材料,通过构建重组转移载体pBOSK△IC-1和pBOSK△IC/ompP2,构成卡那霉素抗性基因(Kanr)和枯草芽胞杆菌(Bacillus subtilis)的果聚蔗糖酶基因(sacB)的正负双向筛选表达盒,并筛选获得SW1株缺失毒素Ⅰ的激活基因C(Apx(I)C),同时嵌合Hps外膜蛋白P2基因(ompP2)的复合突变株SW1 △ApxIC/ompP2.该突变株经鉴定,与SW1亲本株相比,其缺失了大小为475 bp的apxIC基因,同时嵌合有大小为1 107bp的Hps ompP2基因,其基本生长特性与亲本株(SW1)没有显著区别;同时在体外,连续传代10代之后缺失的apxIC基因不会发生回复突变,嵌入的ompP2基因仍能够稳定遗传.本研究成功构建了猪传染性胸膜肺炎放线杆菌缺失apxIC基因并嵌合有Hps ompP2基因复合突变株,为今后研究APP和Hps新型二联疫苗打下基础.%Actinobacillus pkuropnewnoniae (APP) and Haemophilus parasuis (Hps) are the bacteria of causing porcine disease. In this study, We constructed two recombinant vectores pBOSKA/C-1 and pBOSKAIC/ompP2 using APP5 (SW1) strain as a template, and the two recombinant vectores were used for plus-minus screening system with the basis of Kanr and sacB. The mutant strain, named SW1 A apxICIompP2, was obtained, which was deleted toxin I gene C (apxIC) in SW1 and gotnphosised outer membrane protein P2 Hps gene {ompPZ). The mutant missed a size of 475 bp apxIC gene but was inserted a size of 1107 bp Hps ompP2 gene compared with the parental strain SW1. No reverse mutation of apxlC gene was observed during after 10 generations, while Hps ompP2 gene still had a good genetic stability in vitro, the mutant strain had no significant difference of he basic growth characteristics with the parental strain (SW1). The results confirmed that an \\apxlC mutant of A

  5. Actinobacillus pleruropneumoniae transcriptome analysis during early infection - coping with a hostile environment

    Schou, Kirstine Klitgaard; Rundsten, Carsten Friis; Jensen, Tim Kåre;


    Aim: To obtain an increased understanding of how the porcine lung pathogen Actinobacillus pleuropneumoniae (Ap) establish infection in the host. Understanding the means by which a pathogen establishes and maintains infection in the host organism is the first step towards controlling disease...... the next 42 hours. Functional analysis identified a number of putative virulence genes to be initially up-regulated. Conclusions: This is the first study monitoring the development of Ap response in the porcine host during early infection. The ability of pathogenic bacteria to adjust gene expression...... in response to environmental stimuli is critical for bacterial survival within the host. The genes identified as differentially expressed in this study may represent a core set of genes which are mobilized to cope with the host immune response and adapt to the hostile environment. The potential virulence...


    DiRienzo, J M; Slots, J


    Actinobacillus acrinomycetemcomirans isolates from periodontal pockets were examined for restriction fragment-length polymorphism using a characterized 4.7-kb DNA probe. A total of 6 patterns of RFLP was found in 133 isolates originating from 12 subjects. No relatedness was found between RFLP types and serotypes. Different periodontal sites within the same subject and different individuals within the same family sometimes showed only one type of A. actinomycetemcomitans RFLP. When members amo...

  7. Pleural effusion associated with acute and chronic pleuropneumonia and pleuritis secondary to thoracic wounds in horses: 43 cases (1982-1992).

    Collins, M B; Hodgson, D R; Hutchins, D R


    Case records of 43 horses with pleural effusion associated with acute pleuropneumonia, chronic pleuropneumonia, or pleuritis secondary to a penetrating thoracic wound were reviewed to determine the predisposing factors, diagnosis, and treatment of this condition. Acute pleuropneumonia was diagnosed in 36 horses, the majority of which were Thoroughbreds (89%). Of 22 (61%) horses that were in race training at the onset of illness, 11 (31%) had been recently transported a long distance and 4 (11%) had evidence of exercise-induced pulmonary hemorrhage. Physical examination findings and hematologic data were nonspecific. The most consistent abnormality was hyperfibrino-genemia. Affected horses were treated with antibiotics, thoracic drainage, nonsteroidal anti-inflammatory drugs, and supportive care. Twenty-two (61%) horses were discharged from the hospital, with the mean duration of hospitalization for those discharged being 23 days. Nine (25%) horses were euthanatized and 5 (14%) died. Bacterial culturing of thoracic fluid resulted in growth in 30 of the 36 (83%) horses. The finding of anaerobic bacteria in thoracic fluid was not associated with a lower survival rate (62%) than the overall survival rate (61%). Four horses with chronic pleuropneumonia had a history of lethargy and inappetence for > 2 weeks. Actinobacillus equuli was isolated, either alone or in combination with other bacteria, from thoracic fluid of these 4 horses. Each horse was treated with broad spectrum antibiotics and made a rapid recovery. Three horses with acute pleuritis secondary to penetrating thoracic wounds also had nonspecific clinical signs, apart from the wound and a large volume of pleural effusion. Bacteriologic isolates from these horses differed slightly from those of horses with acute pleuropneumonia. PMID:7744650

  8. Clinical significance and taxonomy of Actinobacillus hominis

    Friis-Møller, Alice; Christensen, J J; Fussing, V;


    Clinical findings in 36 immunosuppressed patients with lower respiratory tract infection or bacteremia with Actinobacillus hominis are described. Animal contact was only recorded for three patients; nine patients died despite appropriate antimicrobial treatment. Although infections with this micr...

  9. Putative biomarkers for evaluating antibiotic treatment: an experimental model of porcine Actinobacillus pleuropneumoniae infection

    Lauritzen, B.; Lykkesfeldt, J.; Skaanild, M.T.;


    animals received a single dose of either danofloxacin (2.5 mg/kg) or tiamulin (10 mg/kg). To test the discriminative properties of the biomarkers, the dosage regimens were designed with an expected difference in therapeutic efficacy in favour of danofloxacin. Accordingly, the danofloxacin-treated pigs...... recovered clinically within 24h after treatment, whereas tiamulin-treated animals remained clinically ill until the end of the study, 48 h after treatment. A similar Picture was seen for the biomarkers of infection. During the infection period, plasma C-reactive protein (CRP), interleukin-6 and haptoglobin...... increased, whereas plasma zinc, ascorbic acid and alpha-tocopherol decreased. In the danoffoxacin-treated animals, CRP, interleukin-6, zinc, ascorbic acid and alpha-tocopherol reverted significantly towards normalisation within 24h of treatment. In contrast, signs of normalisation were absent (CRP, zinc and...

  10. Complete Genome Sequence of MIDG2331, a Genetically Tractable Serovar 8 Clinical Isolate of Actinobacillus pleuropneumoniae

    Bossé, JT; Chaudhuri, RR; Li, Y; Leanse, LG; Fernandez Crespo, R; Coupland, P; Holden, MT; Bazzolli, DM; Maskell, DJ; Tucker, AW; Wren, BW; Rycroft, AN; Langford, PR


    Wellcome Trust provided funding to Paul Coupland and Matthew Holden under grant number 098051. Biotechnology and Biological Sci- ences Research Council (BBSRC) provided funding to Janine T. Bosse, Roy R. Chaudhuri, Yanwen Li, Leon G. Leanse, Roberto Fernandez Cre- spo, Paul Coupland, Matthew Holden, Denise Mara Soares Bazzolli, Dun- can J. Maskell, Dan Tucker, Brendan W. Wren, Andrew N. Rycroft, Paul R. Langford, and BraDP1t Consortium under grant numbers BB/ G020744/1, BB/G019177/1, BB/G0192...

  11. Actinobacillus actinomycetemcomitans Y4 capsular-polysaccharide-like polysaccharide promotes osteoclast-like cell formation by interleukin-1 alpha production in mouse marrow cultures.

    Nishihara, T.; Ueda, N; Amano, K; Ishihara, Y; Hayakawa, H.; Kuroyanagi, T; Ohsaki, Y; Nagata, K.; Noguchi, T


    The mechanism of osteoclast-like cell formation induced by periodontopathic bacterium Actinobacillus actinomycetemcomitans Y4 (serotype b) capsular-polysaccharide-like polysaccharide (capsular-like polysaccharide) was examined in a mouse bone marrow culture system. When mouse bone marrow cells were cultured with A. actinomycetemcomitans Y4 capsular-like polysaccharide for 9 days, many multinucleated cells were formed. The multinucleated cells showed several characteristics of osteoclasts, inc...

  12. Clinical significance and taxonomy of Actinobacillus hominis

    Friis-Møller, Alice; Christensen, J J; Fussing, V;


    microorganism seem to be rare, the fact that 37 of 46 strains characterized in this study have been found in Copenhagen indicates that under-reporting may occur. A. hominis is phenotypically relatively homogeneous but can be difficult to differentiate from other Actinobacillus species unless extensive...

  13. Selective medium for isolation of Actinobacillus actinomycetemcomitans.

    Slots, J


    A selective medium, TSBV (tryptic soy-serum-bacitracin-vancomycin) agar, was developed for the isolation of Actinobacillus actinomycetemcomitans, TSBV agar contained (per liter) 40 g of tryptic soy agar, 1 g of yeast extract, 100 ml of horse serum. 75 mg of bacitracin, and 5 mg of vancomycin. The TSBV medium suppressed most oral species and permitted significantly higher recovery of A. actinomycetemcomitans than nonselective blood agar medium. The distinct colonial morphology and positive cat...

  14. Differences in iron acquisition from human haemoglobin among strains of Actinobacillus actinomycetemcomitans

    Hayashida, H.; Poulsen, Knud; Kilian, Mogens


    other strains of the species. None of the strains examined could utilize human transferrin as a source of iron. This was in accordance with the presence of a non-functional tbpA gene, which normally encodes the A subunit of the transferrin-binding-protein complex. Southern blot analysis indicated that......To get a better insight into the physiology of the high-toxic JP2 clone of Actinobacillus actinomycetemcomitans serotype b, which is strongly associated with juvenile periodontitis in adolescents of African descent, the modes of iron acquisition in this clone were examined and compared to those of...... functional duplications of tbpA were not present in the genome. Thus, A. actinomycetemcomitans seems to be in a process of evolution, in which iron acquisition from host transferrin is not essential as in many other members of the pasteurellaceae. All strains could utilize haem as a source of iron. All 11 A...

  15. Fluoroquinolones in the treatment of Actinobacillus actinomycetemcomitans associated periodontitis

    Kleinfelder, JW; Mueller, RF; Lange, DE


    Background: Periodontitis patients harboring Actinobacillus actinmycetemcomitans (Aa) are prime candidates for systemic antibiotic therapy. Besides tetracycline and the combination of metronidazole and amoxicillin the fluoroquinolones are also believed to have antibacterial activity against Aa. The

  16. Pleuroneumonía Equina (Equine Pleuropneumonia

    Aguilera-Tejero, Escolástico


    Full Text Available ResumenLa pleuroneumonía es un problema frecuente en el caballo. Esta enfermedad consiste en colonización bacteriana del parénquima pulmonar, desarrollo de una neumonía o abscesos pulmonares y la consiguiente extensión del proceso hacia la pleura visceral y el espacio pleural provocando pleuritis. Generalmente, su desarrollose asocia con cualquier condición que favorezca la aspiración de secreciones faríngeas o impida su eliminación (transporte, enfermedades víricas, ejercicio extenuante, anestesia general, etc. Los signos clínicos pueden variar según se trate de un problema agudo o crónico, predominando en el primer caso: fiebre, letargia,descarga nasal, tos, intolerancia al ejercicio, disnea y leurodinia. En los casos crónicos suele aparecer fiebre intermitente, pérdida de peso y edema subesternal El diagnóstico se basa fundamentalmente en la ecografía de la región torácica y el análisis microbiológico y citológico de las secreciones traqueales y pleurales. Su tratamiento se centra en antibioterapia sistémica para inhibir el crecimientobacteriano, drenaje del exceso de líquido pleural (en los casos que dificulte la capacidad respiratoria del animal o sea claramente séptico, administración de terapia antiinflamatoria y analgésica y tratamiento de soporte a base de fluidoterapia, oxigenoterapia y broncodilatadores. El pronóstico de la pleuroneumonía es favorable en los casos que se identifican precozmente y reciben tratamientoagresivo, empeorando mucho en casos crónicos o con complicaciones como la laminitis, colitis asociada a antibióticos y trombosis yugular. Las principales secuelas de este proceso incluyen la formación de abscesos pulmonares, fístulas broncopleurales,neumotórax, y pericarditis restrictiva.SummaryPleuropneumonia is a frequent and severe disease in the horse. It is produced by the bacterial colonization of pulmonary parenchyma, development of pneumonia or pulmonary abscesses and subsequent

  17. Cytoplasmic N-Glycosyltransferase of Actinobacillus pleuropneumoniae Is an Inverting Enzyme and Recognizes the NX(S/T) Consensus Sequence*

    Schwarz, Flavio; Fan, Yao-Yun; Schubert, Mario; Aebi, Markus


    N-Linked glycosylation is a frequent protein modification that occurs in all three domains of life. This process involves the transfer of a preassembled oligosaccharide from a lipid donor to asparagine side chains of polypeptides and is catalyzed by the membrane-bound oligosaccharyltransferase (OST). We characterized an alternative bacterial pathway wherein a cytoplasmic N-glycosyltransferase uses nucleotide-activated monosaccharides as donors to modify asparagine residues of peptides and pro...

  18. Molecular characterisation of the early response in pigs to experimental infection with Actinobacillus pleuropneumoniae using cDNA microarrays

    Hedegaard, Jakob; Skovgaard, Kerstin; Mortensen, Shila;


    inflamed lung tissue, genes encoding immune activating proteins and other pro-inflammatory mediators of the innate immune response were found to be up-regulated. Genes encoding different acute phase reactants were found to be differentially expressed in the liver. Conclusion: The obtained results are......-inflamed lung tissue sampled from inoculated animals as well as in liver and tracheobronchial lymph node tissue sampled from three inoculated animals versus two non-inoculated animals. The lung samples were studied using a porcine cDNA microarray with 5375 unique PCR products while liver tissue and...... tracheobronchial lymph node tissue were hybridised to an expanded version of the porcine microarray with 26879 unique PCR products. Results: A total of 357 genes differed significantly in expression between infected and non-infected lung tissue, 713 genes differed in expression in liver tissue from infected versus...

  19. Epidemiological survey of bovine pleuropneumonia in Mali

    Now that rinderpest has been eradicated, contagious bovine pleuropneumonia (CBPP) remains the most important infectious disease of cattle in many tropical African countries including Mali. It is considered as a priority disease by the World Organization for Animal Health, the African Union/Inter-African Bureau for Animal Resources, the Pan-African Programme for the Control of Epizootics (PACE) and the FAO-Emergency Prevention System for Transboundary Animal Diseases (EMPRES). All these institutions and programmes have recognised that the lack of sufficient epidemiological data on this disease is an obstacle to implementing efficient plans for its control. Therefore, research must be conducted in order to generate the accurate data needed to prepare a technically appropriate and well coordinated control programme. The present epidemiological study was conducted in Mali with the aim of evaluating the seroprevalence of the disease in cattle and its geographical distribution. Serum samples collected from 7 628 cattle in different parts of the country were tested for the presence of specific antibody against CBPP by a c-ELISA. In parallel to this serological study, data on CBPP outbreaks and abattoir lung seizures over a 10-year period (1997-2006) were collected. Preliminary results indicate a national seroprevalence rate of 16.28% (1 242/7 628) and a herd seroprevalence rate of 85.18% (161/189). In general, both rates correlated and were relatively higher in the central and southern regions (17.77-28.33%; 89.29-100% respectively) than in the northern and western regions (4.63- 11.89%; 60-83.33% respectively). The total number of outbreaks reported from the field was 121, while the number of suspected lung lesions detected in abattoirs was 12 470. The distribution of these figures varied between the country's regions but did not correlate with the observed seroprevalence rates. (author)

  20. Infectious and rearing-system related risk factors for chronic pleuritis in slaughter pigs

    Enøe, Claes; Mousing, Jan; Schirmer, Anna Luise; Willeberg, Preben


    of the pig, the weight of the carcass, and the herd of origin were also recorded. Individual blood samples were examined for seropositivity for Actinobacillus pleuropneumoniae (AP) serotypes 2, 6, 7, 12, Haemophilus parasuis, Mycoplasma hyopneumoniae (MYC) and swine influenza (SI). Herd...

  1. Development of Contagious Caprine Pleuropneumonia Inactivated Vaccine( M1601 Strain)

    Zhao; Ping; He; Ying; Chu; Yuefeng; Gao; Pengcheng; Zhang; Xuan; Lu; Zhongxin


    Three batches of contagious caprine pleuropneumonia inactivated vaccine( M1601 strain) developed by the laboratory were studied from the aspects of safety,minimum immune dose,immunity duration and storage life. The results showed that the vaccine was safe to goats under different physiological conditions.Regardless of lambs or adult goats,the minimum immune dose was 3 m L,and the immunity duration and the storage life were 6 and 12 months,respectively.

  2. European bluetongue serotype 8

    Drolet, Barbara S.; Reister-Hendricks, Lindsey M.; Podell, Brendan K.; Breitenbach, Jonathan E.; Mcvey, D.S.; Rijn, van Piet A.; Bowen, Richard A.


    Bluetongue virus (BTV) is an orbivirus transmitted by biting midges (Culicoides spp.) that can result in moderate to high morbidity and mortality primarily in sheep and white-tailed deer. Although only 5 serotypes of BTV are considered endemic to the United States, as many as 11 incursive serotyp

  3. Isolation of Actinobacillus suis from a cat's lung

    Daignault, D.; Chouinard, L.; Møller, Kristian;


    Actinobacillus suis has been isolated from the lungs of a 9-month-old cat. The bacterium was characterized biochemically as well as genetically, and its sensitivity profile to different antimicrobial agents was established. The role of this isolate in the cat's condition is discussed....

  4. Serotyping of Clostridium difficile.

    Toma, S.; Lesiak, G; Magus, M; Lo, H L; Delmée, M.


    A total of 246 live Clostridium difficile cultures were serotyped by a slide agglutination technique. Fifteen grouping antisera were produced which serotyped 98% of the cultures (241 of 246). Our results indicated that certain serogroups may have specific pathogenicity. Strains of serogroups A, G, H, K, S1, and S4 were cytotoxigenic and were isolated mainly from adult patients with pseudomembranous colitis or antibiotic-associated diarrhea. Nontoxigenic strains of serogroups D and Cd-5 were i...

  5. A longitudinal study of serological patterns of respiratory infections in nine infected Danish swine herds

    Andreasen, Margit; Nielsen, Jens; Bækbo, Poul; Willeberg, Preben; Bøtner, Anette

    Sixteen litters of seven pigs from each of nine Danish farrow-to-finish herds were followed to investigate the serological patterns caused by natural infection with Mycoplasma hyponeumoniae, Pasteurella multocida toxin and Actinobacillus pleuroneumoniae serotypes 2, 5-7, 12. In seven of the herds....... hyopneumoniae (enzyme-linked immunosorbent assay), P. multocida toxin (enzyme-linked immunosorbent assay) and A. pleuropneumoniae serotypes 2, 5-7, 12 (complement-fixation tests). The most-common pattern (28%) of seroconversion was that of pigs first seroconverting to A. pleuropneumoniae serotype 2, followed by...... seroconversion to M. hyopneumoniae. Each herd had a dominant serotype of A. pleuropneumoniae to which most pigs seroconverted. Seroconversion to the respiratory pathogens occurred mainly in the growing-to-finishing units (8-24 weeks). The risk of seroconversion to the P. multocida toxin was very low (...

  6. Identification of Genes Coding for Exported Proteins of Actinobacillus actinomycetemcomitans

    Mintz, Keith P.; Fives-Taylor, Paula M.


    Random fusions of genomic DNA fragments to a partial gene encoding a signal sequence-deficient bacterial alkaline phosphatase were utilized to screen for exported proteins of Actinobacillus actinomycetemcomitans in Escherichia coli. Twenty-four PhoA+ clones were isolated and sequenced. Membrane localization signals in the form of signal sequences were deduced from most of these sequences. Several of the deduced amino acid sequences were found to be homologous to known exported or membrane-ass...

  7. Adhesion of Actinobacillus actinomycetemcomitans to a human oral cell line.

    Mintz, K. P.; Fives-Taylor, P M


    Two quantitative, rapid assays were developed to study the adhesion of Actinobacillus actinomycetemcomitans, an oral bacterium associated with periodontal disease, to human epithelial cells. The human oral carcinoma cell line KB was grown in microtiter plates, and adherent bacteria were detected by an enzyme-linked immunosorbent assay with purified anti-A. actinomycetemcomitans serum and horseradish peroxidase-conjugated secondary antibody or [3H]thymidine-labeled bacteria. Adhesion was found...

  8. Requirements for invasion of epithelial cells by Actinobacillus actinomycetemcomitans.

    Sreenivasan, P K; Meyer, D H; Fives-Taylor, P M


    Actinobacillus actinomycetemcomitans, an oral bacterium implicated in human periodontal disease, was recently demonstrated to invade cultured epithelial cells (D. H. Meyer, P. K. Sreenivasan, and P. M. Fives-Taylor, Infect. Immun. 59:2719-2726, 1991). This report characterizes the requirements for invasion of KB cells by A. actinomycetemcomitans. The roles of bacterial and host factors were investigated by using selective agents that influence specific bacterial or host cell functions. Inhibi...

  9. Oxidative and nonoxidative killing of Actinobacillus actinomycetemcomitans by human neutrophils.

    Miyasaki, K T; Wilson, M E; Brunetti, A J; Genco, R J


    Actinobacillus actinomycetemcomitans is a facultative gram-negative microorganism which has been implicated as an etiologic agent in localized juvenile periodontitis and in subacute bacterial endocarditis and abscesses. Although resistant to serum bactericidal action and to oxidant injury mediated by superoxide anion (O2-) and hydrogen peroxide (H2O2), this organism is sensitive to killing by the myeloperoxidase-hydrogen peroxide-chloride system (K.T. Miyasaki, M.E. Wilson, and R.J. Genco, In...

  10. Demonstration of Mycoplasma capricolum subsp capripneumoniae and Mycoplasma mycoides subsp mycoides, small colony type in outbreaks of caprine pleuropneumonia in eastern Tanzania

    Kusiluka, L.J.M.; Semuguruka, W.D.; Kazwala, R.R.; Ojeniy, B.; Friis, N.F.


    . The morbidity and mortality rates were 45%-90% and 14%-50%, respectively. The principal pathological lesions were confined to the thoracic cavity and comprised hydrothorax and serofibrinous pleuropneumonia. The histopathological features consisted of a necrotizing fibrinous pleuropneumonia...

  11. Uses of serology for the diagnosis of contagious bovine pleuropneumonia.

    Rurangirwa, F R


    A serological test involves detection of specific changes, induced by a pathogen, in the properties or actions of serum of an infected host. The test may detect the presence in serum of either antibodies to the pathogen (produced by the host) or antigens (i.e. the infecting agent itself and/or its components). The many serological tests which have been developed for the diagnosis of contagious bovine pleuropneumonia (CBPP) are classified into two groups on the basis of this distinction. To date, no single serological test is able to detect all stages of the disease. Thus the choice of serological test (or combination of tests) will depend on the specific aim of the investigation. Meanwhile, a sensitive, specific and simple 'pen-side' test for the diagnosis of all forms of CBPP is still lacking. PMID:8593394

  12. Epidemiological survey of contagious bovine pleuropneumonia (CBPP) in Mali

    Full text: Mali is a major producer of livestock, which plays a predominant role in the economy of the country. It brings about 10% of the GDP, 15% of export incomes and constitutes the main source of incomes for the 30% of the national population. Now that rinderpest is controlled, contagious bovine pleuropneumonia (CBPP) remains the most important infectious disease of cattle not only in Mali, but in most tropical African countries causing great economic losses including mortality, loss of weight, reduced working ability, reduced fertility and indirect costs due to control programs. The disease is characterized by a severe fibrinous exudative pleuropneumonia and is caused by Mycoplasma mycoides subsp. mycoides SC (MmmCS). It is considered a priority disease by the Office of International Epizootie (OIE), the Pan-African Program for the control of epizootics (PACE) and the FAO-Emergency Prevention System for Transboundary Animal Diseases (FAOEMPRES). All these Institutions have recognized that the lack of information concerning the prevalence and the economic impact of the disease is an obstacle to implement any efficient program for its control in Africa. Therefore, research must be conducted in order to generate accurate data on the incidence and distribution of the disease before any appropriate coordinated control program can be defined for this continent. The present epidemiological study was conducted with the aim of studying the seroprevalence of the disease among cattle in Mali, its geographical distribution and its economical impacts. Serum samples collected from 8000 cattle in different parts of the country were tested for MmmSC specific antibody by the competitive ELISA (c-ELISA) and the complement fixation test (CFT). In parallel, data of CBPP cases for the past 10 years and the economic impact of the disease, including production losses and control costs were evaluated. The preliminary results of this study will be presented here. (author)

  13. Genomic characterization of Pasteurella multocida HB01, a serotype A bovine isolate from China.

    Peng, Zhong; Liang, Wan; Liu, Wenjing; Wu, Bin; Tang, Biao; Tan, Chen; Zhou, Rui; Chen, Huanchun


    Pasteurella multocida infects various domestic and feral animals, generally causing clinical disease. To investigate P. multocida disease in cattle, we sequenced the complete genome of P. multocida HB01 (GenBank accession CP006976), a serotype A organism isolated from a cow in China. The genome is composed of a single circular chromosome of 2,416,068 base pairs containing 2212 protein-coding sequences, 6 ribosomal rRNA operons, and 56 tRNA genes. The present study confirms that P. multocida HB01 possesses a more complete metabolic pathway with an intact trichloroacetic acid cycle for anabolism compared with A. pleuropneumoniae and Haemophilus parasuis. This is the first time that this metabolic mechanism of P. multocida has been described. We also identified a full spectrum of genes related to known virulence factors of P. multocida. The differences in virulence factors between strains of different serotypes and origins were also compared. This comprehensive comparative genome analysis will help in further studies of the metabolic pathways, genetic basis of serotype, and virulence of P. multocida. PMID:26827796

  14. Serotyping of Acinetobacter calcoaceticus.

    B. C. DAS; Ayliffe, G


    Serotyping of Acinetobacter calcoaceticus by direct immunofluorescence and a capsule swelling reaction is described. One hundred isolates, including 12 from an outbreak in a neonatal department, were studied. Ninety five per cent of the isolates were typable by immunofluorescence and could be divided into 30 separate types, but 42.1% of typable strains, including 11 from the outbreak, were of one type. Typing results by the capsular swelling reaction generally followed those of immunofluoresc...

  15. A serotype 10 human rotavirus.

    Beards, G; Xu, L.; Ballard, A.; Desselberger, U; McCrae, M A


    Rotaviruses with genome rearrangements isolated from a chronically infected immunodeficient child (F. Hundley, M. McIntyre, B. Clark, G. Beards, D. Wood, I. Chrystie, and U. Desselberger, J. Virol 61:3365-3372, 1987) are the first recognized human isolates of serotype 10. This was shown by both a direct enzyme-linked immunosorbent assay and virus neutralization assays using serotype specific monoclonal antibodies. The serotype was confirmed by sequence analysis of the gene encoding VP7, which...

  16. Actinobacillus suis and Actinobacillus equuli, emergent pathogens of septic embolic nephritis, a new challenge for the swine industry Actinobacillus suis y Actinobacillus equuli, patógenos emergentes de nefritis embólica séptica, un nuevo desafío para la industria porcina

    CE Benavente; IC Fuentealba


    Kidney lesions are an important cause of tissue condemnation in slaughterhouses. In addition to the potential public health implications, organ condemnations have a significant economic impact on the food animal industry. The condition classified broadly as "nephritis" is one of the main causes of tissue condemnation. Embolic nephritis resembling Actinobacillus equuli infection in foals has been recently detected in sows and market hogs. Actinobacillus suis is phenotypically and phylogenetica...

  17. Three new serotypes of Salmonella.

    Sutch, K E; Blackburn, B O


    Three new Salmonella serotypes belonging to Kauffmann's subgenus I (F. Kauffmann, The Bacteriology of Enterobacteriaceae, 1966) were identified. These serotypes were Salmonella brazos 6,14,18:a:e,n,z15, Salmonella midway 6,14,24:d:1,7, and Salmonella balboa 48a, 48b:z41:monophasic.

  18. The Actinobacillus pleuropneumoniae HMW1C-Like Glycosyltransferase Mediates N-Linked Glycosylation of the Haemophilus influenzae HMW1 Adhesin

    Choi, Kyoung-Jae; Grass, Susan; Paek, Seonghee; St. Geme, Joseph W.; Yeo, Hye-Jeong


    The Haemophilus influenzae HMW1 adhesin is an important virulence exoprotein that is secreted via the two-partner secretion pathway and is glycosylated at multiple asparagine residues in consensus N-linked sequons. Unlike the heavily branched glycans found in eukaryotic N-linked glycoproteins, the modifying glycan structures in HMW1 are mono-hexoses or di-hexoses. Recent work demonstrated that the H. influenzae HMW1C protein is the glycosyltransferase responsible for transferring glucose and ...

  19. No overall relationship between average daily weight gain and the serological response to Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae in eight chronically infected Danish swine herds

    Andreasen, Margit; Mousing, Jan; Thomsen, Lars Krogsgård


    The association between the average daily weight gain (from approximately 4 to 20 weeks of age) and the serological responses to respiratory infections was examined in a longitudinal study including 825 pigs from eight chronically infected herds. Pigs were bled every 4th week (starting from......, early or late as compared to the median time (estimated across herds) of seroconversion for the particular pathogen. The variables "gender", "weight at an approximate age of 4 weeks" and "time" (defining the exact length of the follow-up period), were included as explanatory variables, and "litter......" and "herd" were included as explanatory random variables. The individual pig was the unit of concern. The variable defining time at seroconversion was not significantly associated with the average daily weight gain, when evaluating models across all eight herds. The apparent lack of effect could be because...

  20. Validation of putative reference genes for qRT-PCR normalization in tissues and blood from pigs infected with Actinobacillus pleuropneumoniae

    Skovgaard, Kerstin; Mortensen, Shila; Poulsen, K.T.; Angen, Øystein; Heegaard, Peter M. H.

    The quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) is a sensitive and very efficient technique for quantification of gene expression. However, qRT-PCR relies on accurate normalization of gene expression data, as RNA recovery and cDNA synthesis efficiency might va...

  1. Complete Genome Sequence of Mycoplasma mycoides subsp. mycoides T1/44, a Vaccine Strain against Contagious Bovine Pleuropneumonia

    Gourgues, Géraldine; Barré, Aurélien; Beaudoing, Emmanuel; Weber, Johann; Magdelenat, Ghislaine; Barbe, Valérie; Schieck, Elise; Jores, Joerg; Vashee, Sanjay; Blanchard, Alain; Lartigue, Carole


    Mycoplasma mycoides subsp. mycoides is the etiologic agent of contagious bovine pleuropneumonia. We report here the complete genome sequence of the strain T1/44, which is widely used as a live vaccine in Africa. PMID:27081135

  2. Actinobacillus Actinomycetemcomitans y Porphyromonas Gingivales como principales patógenos periodontales

    A Bascones


    Full Text Available Entre las bacterias relacionadas con la enfermedad periodontal, existen dos especies más claramente asociadas a esta enfermedad: Actinobacillus actinomycetemcomitans y Porphyromonas gingivalis. Este trabajo es una revisión bibliográfica sobre estos dos patógenos periodontales, mostrando su origen, prevalencia, distribución, transmisión y respuesta al tratamiento periodontal.Among the bacteria related to periodontal disease, there are two species clearly associated to this disease: Actinobacillus actinomycetemcomitans and Porphyromonas gingiva lis. This paper presents a review of the literature regarding this two periodontal pathogens, and showing their origin, prevalence, distribution, transmission and response to periodontal treatment.

  3. Phylogenetic relationship of equine Actinobacillus species and distribution of RTX toxin genes among clusters

    Kuhnert, Peter; Berthoud, Hélène; Christensen, Henrik; Bisgaard, Magne; Frey, Joachim


    Equine Actinobacillus species were analysed phylogenetically by 16S rRNA gene (rrs) sequencing focusing on the species Actinobacillus equuli, which has recently been subdivided into the non-haemolytic A. equuli subsp. equuli and the haemolytic A. equuli subsp. haemolyticus. In parallel we determined the profile for RTX toxin genes of the sample of strains by PCR testing for the presence of the A. equuli haemolysin gene aqx, and the toxin genes apxI, apxII, apxIII and apxIV, which are known in...

  4. Economical succinic acid production from cane molasses by Actinobacillus succinogenes.

    Liu, Yu-Peng; Zheng, Pu; Sun, Zhi-Hao; Ni, Ye; Dong, Jin-Jun; Zhu, Lei-Lei


    In this work, production of succinic acid by Actinobacillus succinogenes CGMCC1593 using cane molasses as a low cost carbon source was developed. In anaerobic bottles fermentation, succinic acid concentration of 50.6+/-0.9 g l(-1) was attained at 60 h using an optimum medium containing molasses pretreated with sulfuric acid, resulting in a succinic acid yield of 79.5+/-1.1% and sugar utilization of 97.1+/-0.6%. When batch fermentation was carried out in a 5-l stirred bioreactor with pretreated molasses, 46.4 g l(-1) of succinic acid was attained at 48 h and faster cells growth was also observed. Fed batch fermentation was performed to minimize the substrate (sugar) inhibition effect, giving 55.2 g l(-1) of succinic acid and 1.15 g l(-1)h(-1) of productivity at 48 h. The present study suggests that the inexpensive cane molasses could be utilized for the economical and efficient production of succinic acid by A. succinogenes. PMID:17532626

  5. Synergistic effects between amoxicillin, metronidazole, and the hydroxymetabolite of metronidazole against Actinobacillus actinomycetemcomitans.

    Pavicić, M J; van Winkelhoff, A J; de Graaff, J


    Interactions between metronidazole and amoxicillin, metronidazole and its hydroxymetabolite, and amoxicillin and the hydroxymetabolite of metronidazole were investigated with checkerboard titrations in combination with accurately determined MICs and MBCs. Actinobacillus actinomycetemcomitans was used as the test organism. Synergism was found for all three combinations. Fractional inhibitory concentration indices and fractional bactericidal concentration indices varied from 0.3 to 0.7. These s...

  6. Improved diagnosis of contagious bovine pleuropneumonia (CBPP) by standard serological techniques and DNA amplification

    The aim was to improve the diagnosis of contagious bovine pleuropneumonia (CBPP) in Botswana by the implementation and establishment of the OIE standard complement fixation test (CFT) method, and to demonstrate a diagnostic system in a 96 well format coupled with a colorimetric detection of the PCR amplified product suitable for moderate to high sample numbers for the detection of Mycoplasma mycoides mycoides SC (MmmSC) from nasal swabs, lung tissue and cultures

  7. Klebsiella pneumoniae Bacteremia and Capsular Serotypes, Taiwan

    Liao, Chun-Hsing; Huang, Yu-Tsung; Lai, Chih-Cheng; Chang, Cheng-Yu; Chu, Fang-Yeh; Hsu, Meng-Shiuan; Hsu, Hsin-Sui; Hsueh, Po-Ren


    Capsular serotypes of 225 Klebsiella pneumoniae isolates in Taiwan were identified by using PCR. Patients infected with K1 serotypes (41 isolates) had increased community-onset bacteremia, more nonfatal diseases and liver abscesses, lower Pittsburgh bacteremia scores and mortality rates, and fewer urinary tract infections than patients infected with non–K1/K2 serotypes (147 isolates).

  8. Three new serotypes of Rhodococcus equi in Prescott's serotyping system - Short communication.

    Makrai, László; Fodor, László; Hajtós, István; Varga, János; Dénes, Béla


    Three new serotypes were found among Rhodococcus equi strains, which could not be assigned into any of the seven serotypes of Prescott's system. Fortythree R. equi strains out of 44 previously nontypable ones isolated in Hungary could be allocated into one of the three new serotypes using the agar gel immunodiffusion (AGID) test. The three new suggested serotypes are serotype 8 (proposed reference strain: HNCMB-138003), serotype 9 (proposed reference strain: HNCMB-138004) and serotype 10 (proposed reference strain: HNCMB-138005). Hyperimmune sera produced in rabbits against the new serotypes and reference strains gave precipitation only with their homologous antigens, and no crossreactions were observed. All of the previously nontypable isolates from clinical samples of horses (lung abscesses, intestinal lymph nodes, mediastinal lymph nodes) proved to be serotype 8, while strains of serotypes 8, 9 and 10 could be isolated from nasal and rectal swabs of horses and from the soil. Serotype 9 dominated among the previously nontypable strains of swine origin. One of the previously nontypable human strains was serotype 10. This serotype was also isolated from pigs, horses and the soil. The description of the three new serotypes can help us reveal new correlations between the host species, geographical origin and serotype of R. equi isolates. PMID:26551416

  9. Inokulasi Bakteri Selulolitik Actinobacillus sp. Asal Rumen pada Daun Jati Menurunkan Serat Kasar dan Meningkatkan Protein Kasar

    Mirni Lamid


    Full Text Available Constraints use teak leaves as ruminant feed is a high content of crude fiber and low crude protein.The objective of this research was to determine potency of inoculation  Actinobacillus sp. of rumen productionon teak leaves fermentation process could improve quality of teak leaves as animal feed for ruminants.Design study was Completely Randomized Design with four treatments and five replications. Fourtreatment groups were consist of  : P0 = teak leaves + 2% molasses (control; P1 = teak leaves + 2%molasses + 5% Actinobacillus sp.;  P2 = teak leaves + 2% molasses + 10% Actinobacillus sp.;  P3 = teakleaves + 2% molasses + 15%  Actinobacillus sp. Proximate analysis were done after teak leaves werefermented for seven days. The data were analyzed with Analysis of Variance followed by Duncan’s MultipleRange Test. The results showed a significant difference (P <0.05 for crude fiber and crude protein P3 andP2 compared with control’s. Crude fiber content of P3 and P2 respectively 31.67% and 32%, while the crudeprotein content of P3 and P2 respectively 13.57% and 13.31%. Conclusions of this research are:  InoculationActinobacillus sp. with doses 5%, 10% and 15% in the fermentation teak leaves can decrease crude fibercontent and increased crude protein content. Dose efficient to use bacterial fermentation teak leavesActinobacillus sp. is 10%.

  10. New Salmonella serotype: Salmonella enteritidis serotype Grandhaven (30(1):r:1,2).

    McDougal, D L; Treleaven, B E; Renshaw, E C


    A new Salmonella serotype, Salmonella enteritidis serotype Grandhaven (30(1):r:1,2), was isolated from the stool of a 35-year-old man with mild gastroenteritis. He had just returned from Sudan, Africa.

  11. Porphyromonas gingivalis, Prevotella intermedia and Actinobacillus actinomycetemcomitans in the Plaque of Children without Periodontitis

    W. Zimmer; Wilson, M.; Marsh, P D; Newman, H.N.; Bulman, J.


    The aim of this study was to find out whether the suspected periodontal pathogens Actinobacillus actinomycetemcomitans, Prevotella intermedia and Porphyromonas gingivalis could be recovered from the dental plaque and the dorsum of the tongue in children without periodontal breakdown. Thirty-six male Caucasian children participated in this study, 21 aged 6-11 y and 15 aged 14-16 y. Subcontact area plaque and tongue samples were cultured on selective media and a presumptive identification of th...

  12. Acquisition and Colonization Stability of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in Children

    Lamell, Celeste W.; Griffen, Ann L.; McClellan, Dawn L.; Eugene J Leys


    The presence of Porphyromonas gingivalis has been shown to be a risk factor for periodontitis in adults, and Actinobacillus actinomycetemcomitans has been implicated as a pathogen in early-onset periodontitis. Both species have been shown to establish stable colonization in adults. In cross-sectional studies, both A. actinomycetemcomitans and P. gingivalis have been detected in over one-third of apparently healthy children. Information on the stability of colonization with these organisms in ...

  13. PCR Methods for Rapid Identification and Characterization of Actinobacillus seminis Strains

    Appuhamy, S; Coote, J G; Low, J. C.; Parton, R


    Twenty-four isolates of Actinobacillus seminis were typed by PCR ribotyping, repetitive extragenic palindromic element (REP)-based PCR, and enterobacterial repetitive intergenic consensus (ERIC)-based PCR. Five types were distinguished by REP-PCR, and nine types were distinguished by ERIC-PCR. PCR ribotyping produced the simplest pattern and could be useful for identification of A. seminis and for its differentiation from related species. REP- and ERIC-PCR could be used for strain differentia...

  14. Evidence for invasion of a human oral cell line by Actinobacillus actinomycetemcomitans.

    Meyer, D H; Sreenivasan, P K; Fives-Taylor, P M


    Actinobacillus actinomycetemcomitans, an oral bacterial species associated with periodontal disease, was found to invade human cell lines. Invasion was demonstrated by recovery of viable organisms from gentamicin-treated KB cell monolayers and by light and electron microscopy. Internalization occurred through a cytochalasin D-sensitive process. Invasion efficiencies of some A. actinomycetemcomitans strains were comparable to those of invasive members of the family Enterobacteriaceae. Differen...

  15. Actinobacillus actinomycetemcomitans in Human Periodontal Disease: a Cross-Sectional Microbiological Investigation

    Slots, Jørgen; Reynolds, Homer S.; Genco, Robert J.


    Actinobacillus actinomycetemcomitans is a facultative gram-negative bacterium which has been associated with severe oral and nonoral infections. This study examined its occurrence in the oral cavities of 10 normal juveniles, 11 normal adults, 10 juvenile periodontitis patients, and 12 adult periodontitis patients. Four deep periodontal pockets and two normal periodontal sites were sampled in the diseased patients, and six normal periodontal sites were sampled in the healthy individuals. In al...

  16. Actinobacillus Actinomycetemcomitans y Porphyromonas Gingivales como principales patógenos periodontales

    A Bascones; A Caballeros


    Entre las bacterias relacionadas con la enfermedad periodontal, existen dos especies más claramente asociadas a esta enfermedad: Actinobacillus actinomycetemcomitans y Porphyromonas gingivalis. Este trabajo es una revisión bibliográfica sobre estos dos patógenos periodontales, mostrando su origen, prevalencia, distribución, transmisión y respuesta al tratamiento periodontal.Among the bacteria related to periodontal disease, there are two species clearly associated to this disease: Actinobacil...

  17. Autoinducer 2 Is Required for Biofilm Growth of Aggregatibacter (Actinobacillus) actinomycetemcomitans▿

    Shao, HanJuan; Lamont, Richard J.; Demuth, Donald R.


    Autoinducer 2 (AI-2) is required for the growth of Aggregatibacter (Actinobacillus) actinomycetemcomitans in culture under conditions of iron limitation. However, in vivo this organism thrives in a complex multispecies biofilm that forms in the human oral cavity. In this report, we show that adherent growth of A. actinomycetemcomitans on a saliva-coated surface, but not planktonic growth under iron-replete conditions, is defective in a LuxS-deficient background. Biofilm growth of the luxS mut...

  18. Effect of Ciprofloxacin on Killing of Actinobacillus actinomycetemcomitans by Polymorphonuclear Leukocytes

    Cacchillo, David A.; Walters, John D.


    Actinobacillus actinomycetemcomitans, a pathogen associated with aggressive periodontitis, resists phagocytic killing by polymorphonuclear leukocytes (PMNs). It is susceptible to ciprofloxacin, which PMNs actively accumulate. This study tested the hypothesis that ciprofloxacin-loaded PMNs are more effective at killing A. actinomycetemcomitans than control PMNs. Isolated human PMNs were loaded by brief incubation with 0.5 μg of ciprofloxacin/ml. Opsonized bacteria (ATCC 43718) were incubated a...

  19. Actinobacillus suis and Actinobacillus equuli, emergent pathogens of septic embolic nephritis, a new challenge for the swine industry Actinobacillus suis y Actinobacillus equuli, patógenos emergentes de nefritis embólica séptica, un nuevo desafío para la industria porcina

    CE Benavente


    Full Text Available Kidney lesions are an important cause of tissue condemnation in slaughterhouses. In addition to the potential public health implications, organ condemnations have a significant economic impact on the food animal industry. The condition classified broadly as "nephritis" is one of the main causes of tissue condemnation. Embolic nephritis resembling Actinobacillus equuli infection in foals has been recently detected in sows and market hogs. Actinobacillus suis is phenotypically and phylogenetically closely related to A. equuli. Both are Gram-negative bacteria, not easy to detect in routine exams. A. suis is an opportunistic pathogen that can produce fatal septicaemia in pigs, pneumonia, polyarthritis, septic embolic nephritis, abortion and mummified foetuses. Outbreaks of clinical disease appear to occur more frequently in high-health-status herds. In adult pigs the skin lesions may be confused with porcine erysipelas. A. suis and A. equuli are emerging opportunistic pathogens in the porcine industry and both have potential public health consequences to people that handles meat products. The objective of this paper is to present a literature review regarding the role of A. suis and A. equuli in the pathogenesis of nephritis in swine.Las lesiones renales son una causa importante de decomiso en los mataderos. Además de las posibles consecuencias en salud pública, el decomiso de órganos tiene un gran impacto económico en la industria de alimento animal. Recientemente, nefritis embólica séptica con lesiones semejantes a infecciones con Actinobacillus equuli en potrillos ha sido detectada en reproductoras y cerdos con peso de mercado. Actinobacillus equuli es fenotípica y genéticamente similar a Actinobacillus suis. Ambas son bacterias Gram-negativas difíciles de diagnosticar en exámenes de rutina. A. suis es un patógeno oportunista capaz de producir septicemia en cerdos, neumonía, poliartritis, nefritis embólica séptica, aborto y fetos

  20. Final classification of Bisgaard taxon 9 as Actinobacillus arthritidis sp nov and recognition of a novel genomospecies for equine strains of Actinobacillus lignieresii

    Christensen, Henrik; Bisgaard, Magne; Angen, Øystein; Olsen, John Elmerdahl


    Phenotypic characterization of bacteria from diseased and healthy horses identified 18 isolates as Bisgaard taxon 9 and 11 isolates as Actinobacillus lignieresii. All strains of taxon 9 were alpha-galactosidase- and raffinose-positive and showed variable fermentation of (+)L-arabinose and (-)D-sorbitol....... Strains of A. lignieresii were negative for these characteristics, with the exception of raffinose. Two strains from the (-)D-sorbitol-negative group of taxon 9 showed a 16S rRNA similarity of 99.6%, while 99.5% similarity was found between two strains of the (-)D-sorbitol-positive group. DNA......-DNA hybridization between the two strains representing the (-)D-sorbitol-negative group showed 98% binding, and their closest relationship was to a strain of A. lignieresii (64%). The two strains of the (-)D-sorbitol-positive group showed 83% binding and were related to the (-)D-sorbitol-negative group at a 76% DNA...

  1. luxS and arcB Control Aerobic Growth of Actinobacillus actinomycetemcomitans under Iron Limitation

    Fong, Karen P.; Gao, Ling; Demuth, Donald R.


    LuxS is responsible for the production of autoinducer 2 (AI-2), which functions in Vibrio harveyi as a quorum-sensing signal that controls the cell density-dependent expression of the lux operon. In nonluminescent organisms, the physiologic role of AI-2 is not clear. We report that inactivation of luxS in Actinobacillus actinomycetemcomitans JP2 results in reduced growth of the mutant, but not the wild-type organism, under aerobic, iron-limited conditions. Stunted cultures of the luxS mutant ...

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

    Soyer, Yeşim; Orsi, Renato H.; Rodriguez-Rivera, Lorraine D; Sun, Qi; Wiedmann, Martin


    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 (Typhimur...

  3. Using Participatory Epidemiology Tools to Investigate Contagious Caprine Pleuropneumonia (CCPP in Maasai Flocks, Northern Tanzania

    Emmanuel Senyael


    Full Text Available Participatory Epidemiology (PE was applied on the Maasai rangeland of northern Tanzania to understand pastoralist’s perceptions of the clinical and epidemiological features of Contagious Caprine Pleuropneumonia (CCPP. The study was conducted during the period of April 2008 and caprine disease event was directed during the period of July 2006 to June 2007. Participatory methods such as Focus Group Discussion (FGD, proportional piling and matrix scoring were used to characterize pastoralist perceived clinical signs and risk factors for CCPP. The estimated mean incidence and case mortality rate of CCPP was 31.6 and 61.4%, respectively. Matrix scoring showed moderate to good agreement between informant groups on the clinical signs and risk factors. It was concluded that PE complimented with local knowledge could generally be used to generate disease information at low cost and therefore assist the design of feasible disease surveillance systems and control programmes at local and national level.

  4. Diagnosis of contagious caprine and contagious bovine pleuropneumonia by PCR and restriction enzyme analysis

    Contagious caprine pleuropneumonia (CCPP) and contagious bovine pleuropneumonia (CBPP) are caused by Mycoplasma capricolum ssp. capripneumoniae and Mycoplasma mycoides ssp. mycoides, small colony type (SC), respectively. These species belong to the group of six mycoplasmas referred to as the Mycoplasma mycoides cluster. The members of the M. mycoides cluster are closely related, and some of them are very difficult to grow. Diagnosis based on identification of the causative agents by cultivation, biochemical reactions, or serology is therefore difficult, and improved diagnostic methods are sorely needed. Identification methods for M. capricolum ssp. capripneumoniae and M. mycoides ssp. mycoides SC, based on detection of the 16S rRNA genes from all members of the M. mycoides cluster by polymerase chain reaction (PCR) and differentiation of species by restriction enzyme analysis with PstI and AluI, respectively, have been developed in the Department of Bacteriology, National Veterinary Institute Laboratory, Sweden. The methods are based on the fact that members of the M. mycoides cluster have two rRNA operons and that there are sequence differences (polymorphisms) between the two 16S rRNA genes from the two operons. The majority of these polymorphisms are characteristic for each species, and some of them can be utilized for differentiation between the species. Techniques for extraction of DNA from lung tissue and from clinical material dried onto filter paper for preservation are described. Pleural fluid was found to be an ideal sample for direct analysis by PCR, but this material can also be dried onto filter paper for preservation and sent to another laboratory for the PCR analysis. These identification systems designed for M. capricolum ssp. capripneumoniae and M. mycoides ssp. mycoides SC have been used for diagnosis of CCPP and CBPP, respectively, by analysis of clinical samples brought to the laboratory as frozen lung tissue or preserved by drying onto a filter

  5. Serotype identification of Cryptococcus neoformans by multiplex PCR.

    Ito-Kuwa, S; Nakamura, K; Aoki, S; Vidotto, V


    The pathogenic yeast Cryptococcus neoformans is traditionally classified into three varieties with five serotypes: var. grubii (serotype A), var. neoformans (serotype D), var. gattii (serotypes B and C) and serotype AD (hybrid of serotypes A and D). A commercial kit, Crypto Check (Iatron Laboratories, Tokyo, Japan), has been used worldwide for serotyping isolated strains. However, its production was discontinued in 2004, and hence the present study aimed to develop a simple polymerase chain reaction (PCR) method for serotyping C. neoformans strains. Subjecting genomic DNA of 59 strains of the five serotypes to multiplex PCR amplification using a set of four primers designed for the laccase gene (LAC1) differentiated serotypes A, D, B and C, but could not separate serotype AD from serotype D. However, a primer pair designed for the capsule gene (CAP64) allowed serotypes D and AD to be differentiated. When PCR amplification was performed in the simultaneous presence of the above six primers, the five serotypes produced two to five DNA fragments that could be used to distinguish them. This multiplex PCR method is useful for serotyping C. neoformans isolates, and represents an effective replacement for the Crypto Check kit. PMID:17576319

  6. An international collaborative study to determine the prevalence of contagious caprine pleuropneumonia by monoclonal antibody-based cELISA

    Peyraud, Armelle; Poumarat, Francois; Tardy, Florence; Manso-Silvan, Lucia; Hamroev, Karomatullo; Tilloev, Tillo; Amirbekov, Mullojon; Tounkara, Karim; Bodjo, Charles; Wesonga, Hezron; Nkando, Isabel Gacheri; Jenberie, Shiferaw; Yami, Martha; Cardinale, Eric; Meenowa, Deodass


    Background: Few serological tests are available for detecting antibodies against Mycoplasma capricolum subsp. capripneumoniae, the causal agent of contagious caprine pleuropneumonia (CCPP). The complement fixation test, the test prescribed for international trade purposes, uses a crude antigen that cross-reacts with all the other mycoplasma species of the "mycoides cluster" frequently infecting goat herds. The lack of a more specific test has been a real obstacle to the evaluation of the prev...

  7. Production of succinic acid from oil palm empty fruit bunch cellulose using Actinobacillus succinogenes

    Pasma, Satriani Aga; Daik, Rusli; Maskat, Mohamad Yusof


    Succinic acid is a common metabolite in plants, animals and microorganisms. It has been used widely in agricultural, food and pharmaceutical industries. Enzymatic hydrolysate glucose from oil palm empty fruit bunch (OPEFB) cellulose was used as a substrate for succinic acid production using Actinobacillus succinogenes. Using cellulose extraction from OPEFB can enhance the production of glucose as a main substrate for succinic acid production. The highest concentration of glucose produced from enzymatic hydrolysis is 167 mg/mL and the sugar recovery is 0.73 g/g of OPEFB. By optimizing the culture medium for succinic acid fermentation with enzymatic hydrolysate of OPEFB cellulose, the nitrogen sources could be reduced to just only 2.5 g yeast extract and 2.5 g corn step liquor. Batch fermentation was carried out using enzymatic hydrolysate of OPEFB cellulose with yeast extract, corn steep liquor and the salts mixture, 23.5 g/L succinic acid was obtained with consumption of 72 g/L glucose in enzymatic hydrolysate of OPEFB cellulose at 38 hours and 37°C. This study suggests that enzymatic hydrolysate of OPEFB cellulose maybe an alternative substrate for the efficient production of succinic acid by Actinobacillus succinogenes.

  8. Multi-serotype pneumococcal nasopharyngeal carriage prevalence in vaccine naive Nepalese children, assessed using molecular serotyping.

    Rama Kandasamy

    Full Text Available Invasive pneumococcal disease is one of the major causes of death in young children in resource poor countries. Nasopharyngeal carriage studies provide insight into the local prevalence of circulating pneumococcal serotypes. There are very few data on the concurrent carriage of multiple pneumococcal serotypes. This study aimed to identify the prevalence and serotype distribution of pneumococci carried in the nasopharynx of young healthy Nepalese children prior to the introduction of a pneumococcal conjugate vaccine using a microarray-based molecular serotyping method capable of detecting multi-serotype carriage. We conducted a cross-sectional study of healthy children aged 6 weeks to 24 months from the Kathmandu Valley, Nepal between May and October 2012. Nasopharyngeal swabs were frozen and subsequently plated on selective culture media. DNA extracts of plate sweeps of pneumococcal colonies from these cultures were analysed using a molecular serotyping microarray capable of detecting relative abundance of multiple pneumococcal serotypes. 600 children were enrolled into the study: 199 aged 6 weeks to <6 months, 202 aged 6 months to < 12 months, and 199 aged 12 month to 24 months. Typeable pneumococci were identified in 297/600 (49.5% of samples with more than one serotype being found in 67/297 (20.2% of these samples. The serotypes covered by the thirteen-valent pneumococcal conjugate vaccine were identified in 44.4% of samples containing typeable pneumococci. Application of a molecular serotyping approach to identification of multiple pneumococcal carriage demonstrates a substantial prevalence of co-colonisation. Continued surveillance utilising this approach following the introduction of routine use of pneumococcal conjugate vaccinates in infants will provide a more accurate understanding of vaccine efficacy against carriage and a better understanding of the dynamics of subsequent serotype and genotype replacement.

  9. Transplacental Transmission of Bluetongue Virus Serotype 1 and Serotype 8 in Sheep: Virological and Pathological Findings

    Sluijs, van der M.T.W.; Schroer-Joosten, D.P.H.; Fid-Fourkour, A.; Vrijenhoek, M.P.; Debyser, I.; Moulin, V.; Moormann, R.J.M.; Smit, de A.J.


    The Bluetongue virus serotype 8 (BTV-8) strain, which emerged in Europe in 2006, had an unusually high ability to cause foetal infection in pregnant ruminants. Other serotypes of BTV had already been present in Europe for more than a decade, but transplacental transmission of these strains had never


    Aren, Gamze; Aktören, Oya; Külekçi, Güven


    ÖzetActinobacillus actinomycetemcomitans lokalİze jüve-nil periodontitis, Papjllon- Le fevre sendromunun bazı olguları ve erişkinlerde hızlı ilerleyen periodontitiste başlıca sorumlu patojendir. Bununla birlikte normal ağız florasının da bir yerleşiği olarak bulunabilir. Bu çalışmada 4-7 yaşlan arasındaki sağlıklı 16 çocukta süt diş dizinde A.actinomycetemcomitans'ıun varbğı araştırılmıştır. Bu amaçla her çocuktan alt ikinci süt azısının mesialinden diş ipi ile plak ve dil sırtından bakteriyo...

  11. Serotype distribution in non-bacteremic pneumococcal pneumonia

    Benfield, Thomas; Skovgaard, Marlene; Schønheyder, Henrik Carl;


    There is limited knowledge of serotypes that cause non-bacteremic pneumococcal pneumonia (NBP). Here we report serotypes, their associated disease potential and coverage of pneumococcal conjugate vaccines (PCV) in adults with NBP and compare these to bacteremic pneumonia (BP)....

  12. Succinic acid production from duckweed (Landoltia punctata) hydrolysate by batch fermentation of Actinobacillus succinogenes GXAS137.

    Shen, Naikun; Wang, Qingyan; Zhu, Jing; Qin, Yan; Liao, Siming; Li, Yi; Zhu, Qixia; Jin, Yanling; Du, Liqin; Huang, Ribo


    Duckweed is potentially an ideal succinic acid (SA) feedstock due to its high proportion of starch and low lignin content. Pretreatment methods, substrate content and nitrogen source were investigated to enhance the bioconversion of duckweed to SA and to reduce the costs of production. Results showed that acid hydrolysis was an effective pretreatment method because of its high SA yield. The optimum substrate concentration was 140g/L. The optimum substrate concentration was 140g/L. Corn steep liquor powder could be considered a feasible and inexpensive alternative to yeast extract as a nitrogen source. Approximately 57.85g/L of SA was produced when batch fermentation was conducted in a 1.3L stirred bioreactor. Therefore, inexpensive duckweed can be a promising feedstock for the economical and efficient production of SA through fermentation by Actinobacillus succinogenes GXAS137. PMID:27023386

  13. Analysis of Salmonella enterica Serotype-Host Specificity in Calves: Avirulence of S. enterica Serotype Gallinarum Correlates with Bacterial Dissemination from Mesenteric Lymph Nodes and Persistence In Vivo

    Paulin, Susan M.; Watson, Patricia R.; Benmore, Annette R.; Stevens, Mark P.; Jones, Philip W.; Villarreal-Ramos, Bernardo; Wallis, Timothy S.


    Host and bacterial factors that determine whether Salmonella serotypes remain restricted to the gastrointestinal tract or penetrate beyond the mucosa and cause systemic disease remain largely undefined. Here, factors influencing Salmonella host specificity in calves were assessed by characterizing the pathogenesis of different serotypes. Salmonella enterica serotype Dublin was highly virulent intravenously, whereas S. enterica serotype Choleraesuis was moderately virulent. Both serotypes were...

  14. Capsular polysaccharide from Mycoplasma mycoides subsp. mycoides shows potential for protection against contagious bovine pleuropneumonia.

    Mwirigi, Martin; Nkando, Isabel; Olum, Moses; Attah-Poku, Samuel; Ochanda, Horace; Berberov, Emil; Potter, Andrew; Gerdts, Volker; Perez-Casal, Jose; Wesonga, Hezron; Soi, Reuben; Naessens, Jan


    Contagious Bovine Pleuropneumonia (CBPP) is a severe respiratory disease caused by Mycoplasma mycoides subsp. mycoides (Mmm) which is widespread in Africa. The capsule polysaccharide (CPS) of Mmm is one of the few identified virulence determinants. In a previous study, immunization of mice against CPS generated antibodies, but they were not able to prevent multiplication of Mmm in this model animal. However, mice cannot be considered as a suitable animal model, as Mmm does not induce pathology in this species. Our aim was to induce antibody responses to CPS in cattle, and challenge them when they had specific CPS antibody titres similar or higher than those from cattle vaccinated with the live vaccine. The CPS was linked to the carrier protein ovalbumin via a carbodiimide-mediated condensation with 1-ethyl-3(3-imethylaminopropyl) carbodiimide (EDC). Ten animals were immunized twice and challenged three weeks after the booster inoculation, and compared to a group of challenged non-immunized cattle. When administered subcutaneously to adult cattle, the vaccine elicited CPS-specific antibody responses with the same or a higher titre than animals vaccinated with the live vaccine. Pathology in the group of immunized animals was significantly reduced (57%) after challenge with Mmm strain Afadé compared to the non-immunized group, a figure in the range of the protection provided by the live vaccine. PMID:27496744

  15. Challenges of controlling contagious bovine pleuropneumonia in sub-Saharan Africa: a Zambian perspective.

    Muuka, Geoffrey; Songolo, Nadi; Kabilika, Swithine; Hang'ombe, Bernard M; Nalubamba, King S; Muma, John B


    Contagious bovine pleuropneumonia (CBPP) is a disease of economic importance that is widely distributed in sub-Saharan African and contributes significantly to cattle morbidity and mortality. Control of CBPP offers a number of challenges as a result many developing countries in Africa are still struggling with this disease. In this study, we look at the challenges encountered in CBPP control in sub-Saharan Africa from the Zambian perspective. In conducting this study, we reviewed scientific literature and reports from the Ministry of Agriculture and Livestock and related animal institutions, and also made interviews with experts and key government officials involved in CBPP control in Zambia. Among the challenges identified for the successful control of CBPP were as follows: failure in the delivery of veterinary services, lack of a cattle identification system, natural phenomenon, livestock husbandry systems in the traditional sector, human movements, traditional practices among cattle farmers and cattle marketing systems. It was seen that the epidemiology of CBPP in Zambia is influenced by both ecological and anthological factors. Therefore, design and implementation of any control or eradication programme should be area/regional-dependent taking into account the different factors influencing disease transmission and maintenance. PMID:22843213

  16. Pathogenesis and Immunohistochemical Studies of Caprine Pleuropneumonia in Experimentally Infected Goats

    Umer Sadique*, Zafar Iqbal Chaudhry1, Muhammad Younus Rana2, Aftab Ahmad Anjum3, Zahoor-Ul-Hassan, Abdul Sajid and Muhammad Mushtaq


    Full Text Available This study was designed to evaluate the pathogenesis of caprine pleuropneumonia (CPP in the experimentally inoculated goats with Mycoplasma mycoides subspecies Capri (Mmc. For this purpose, 12 goats (Group B were inoculated with bacterial isolates of Mmc while four goats were kept as untreated control (Group A. Clinical signs of the disease were recorded twice daily. Two goats from group B were sacrificed on weekly basis to demonstrate gross pathological lesions in different organs. Tissue samples from lungs, trachea, liver, heart, kidney, spleen, and small intestines were preserved for histopathological studies. The lungs and lymph nodes were preserved to demonstrate the antigen in tissue by using immuno- histochemical technique. The disease was successfully reproduced in all infected goats with severe manifestation. The clinical signs and gross lesions of the disease were mild at the beginning and became severe at the third and fourth weeks and then progressed to moderate and chronic forms. The histopathological lesions characteristic of CPP were found in all the organs. Antigen of Mmc was detected in tissue sections of lungs and lymph nodes. In conclusion, the disease was efficiently reproduced in experimental animals that showed acute septicemic form with lethal outcome.

  17. Contagious bovine pleuropneumonia: Seroprevalence and risk factors in Western Oromia, Ethiopia.

    Daniel, Garuma; Abdurahaman, Mukarim; Tuli, Getachew; Deresa, Benti


    Contagious bovine pleuropneumonia (CBPP) is one of the most important threats to cattle health and production in Ethiopia. At the livestock farm of the Bako Agricultural Research Center, an outbreak of respiratory disease of cattle occurred in May 2011, and many animals were affected and died before the disease was diagnosed. Therefore, this study was designed to determine the seroprevalence of CBPP antibodies in selected districts of Western Oromia Region and to assess the potential risk factors for the occurrence of the disease. A crosssectional study was conducted from November 2013 to March 2014 in three selected districts of Western Oromia Region. A total of 386 sera were examined for the presence of specific antibodies against Mycoplasma mycoidesmycoides small colony (MmmSC), using a competitive enzyme-linked immunosorbent assay. The risk factors that were evaluated in this study were geographical location, age, sex, breed and body condition. The overall seroprevalence in this study was 28.5%. The seroprevalence of Mycoplasma mycoidesmycoides small colony antibodies at the district level was 40.3%, 19.0% and 5.7% in Gobbu Sayyo, BakoTibbe and Horro districts, respectively. There was a statistically significant variation ( p 0.05) with the serological status of the animal. This study showed that the overall prevalence of CBPP in Western Oromia Zones was high. This warrants the implementation of appropriate preventive and control measures to minimise the economic losses associated with the disease. PMID:27247066

  18. Relationship Between Conversion of Localized Juvenile Periodontitis-Susceptible Children From Health to Disease and Actinobacillus actinomycetemcomitans Leukotoxin Promoter Structure*

    Bueno, Lina C.; Mayer, Marcia P. A.; DiRienzo, Joseph M.


    The periodontal pathogen Actinobacillus actinomycetemcomitans produces a leukotoxin that is considered a primary virulence factor in localized juvenile Periodontitis (LJP). Select strains of the bacterium contain a 530-bp deletion in the promoter region of the leukotoxin gene operon which results in enhanced transcription of the leukotoxin. DNA hybridization and polymerase chain reaction (PCR) were used to examine genetic variants of A. actinomycetemcomitans in 24 LJP-susceptible children fro...

  19. Use of 16S rRNA Sequencing for Identification of Actinobacillus ureae Isolated from a Cerebrospinal Fluid Sample

    Whitelaw, A. C.; Shankland, I. M.; Elisha, B. G.


    Actinobacillus ureae, previously Pasteurella ureae, has on rare occasions been described as a cause of human infection. Owing to its rarity, it may not be easily identified in clinical microbiology laboratories by standard tests. This report describes a patient with acute bacterial meningitis due to A. ureae. The identity of the isolate was determined by means of DNA sequence analysis of a portion of the 16S rRNA gene.

  20. Inokulasi Bakteri Selulolitik Actinobacillus sp. Asal Rumen pada Daun Jati Menurunkan Serat Kasar dan Meningkatkan Protein Kasar

    Mirni Lamid; Anggun Foetus Eka Julita; Ngakan Made Rai Widjaya


    Constraints use teak leaves as ruminant feed is a high content of crude fiber and low crude protein.The objective of this research was to determine potency of inoculation  Actinobacillus sp. of rumen productionon teak leaves fermentation process could improve quality of teak leaves as animal feed for ruminants.Design study was Completely Randomized Design with four treatments and five replications. Fourtreatment groups were consist of  : P0 = teak leaves + 2% molasses (control); P1 = teak lea...

  1. Fatal meningitis in a previously healthy young adult caused by Streptococcus pneumoniae serotype 38: an emerging serotype?

    Pearse Lisa A


    Full Text Available Abstract Background In December 2001, a fatal case of pneumococcal meningitis in a Marine Corps recruit was identified. As pneumococcal vaccine usage in recruit populations is being considered, an investigation was initiated into the causative serotype. Case presentation Traditional and molecular methods were utilized to determine the serotype of the infecting pneumococcus. The pneumococcal isolate was identified as serotype 38 (PS38, a serotype not covered by current vaccine formulations. The global significance of this serotype was explored in the medical literature, and found to be a rare but recognized cause of carriage and invasive disease. Conclusion The potential of PS38 to cause severe disease is documented in this report. Current literature does not support the hypothesis that this serotype is increasing in incidence. However, as we monitor the changing epidemiology of pneumococcal illness in the US in this conjugate era, PS38 might find a more prominent and concerning niche as a replacement serotype.

  2. Emerging resistant serotypes of invasive Streptococcus pneumoniae

    Elshafie, Sittana; Taj-Aldeen, Saad J


    Background Streptococcus pneumoniae is the leading cause of meningitis and sepsis. The aim of the study was to analyze the distribution, vaccine serotype coverage, and antibiotic resistance of S. pneumoniae serotypes isolated from patients with invasive diseases, after the introduction of pneumococcal 7-valent conjugated vaccine (PCV-7). Methods A total of 134 isolates were collected from blood and cerebrospinal fluid specimens at Hamad Hospital during the period from 2005 to 2009. Isolate serotyping was done using the Quellung reaction. The prevaccination period was considered before 2005. Results The most common serotypes for all age groups were 3 (12.70%), 14 (11.90%), 1 (11.90%), 19A (9.00%), 9V (5.20%), 23F (5.20%), and 19F (4.50%). Coverage rates for infant conjugated vaccine (PCV-10), and the 13-valent conjugated vaccine (PCV-13) were 34.78%, 52.17%, and 78.26%, respectively. Coverage rates of these vaccines were 50%, 67.86%, and 75% for the 2–5 years age group; 27.12%, 40.68%, and 64.41% for the age group 6–64 years; and 25%, 33.33%, and 66.67% for the ≥65 years age group, respectively. The percentage of nonsusceptible isolates to penicillin, cefotaxime, and erythromycin were 43.86%, 16.66%, and 22.81%, respectively. Thirty-seven isolates (32.46%) were multidrug resistant (MDR) and belonged to serotypes 14, 19A, 19F, 23F, 1, 9V, 12F, 4, 6B, 3, and 15A. Compared to previous results before the introduction of PCV-7, there was a significant reduction in penicillin-nonsusceptable S. pneumoniae from 66.67% to 43.86%, and a slight insignificant reduction in erythromycin nonsusceptible strains from 27.60% to 22.8%, while there was a significant increase in cefotaxime nonsusceptible strains from 3.55% to 16.66%. Conclusion Invasive pneumococcal strains and the emergence of MDR serotypes is a global burden that must be addressed through multiple strategies, including vaccination, antibiotic stewardship, and continuous surveillance. PMID:27418844

  3. A meta-analysis of contagious caprine pleuropneumonia (CCPP) in Ethiopia.

    Asmare, Kassahun; Abayneh, Takele; Mekuria, Solomon; Ayelet, Gelagay; Sibhat, Berhanu; Skjerve, Eystein; Szonyi, Barbara; Wieland, Barbara


    This systematic literature review was initiated due to lack of comprehensive information on the status and distribution of contagious caprine pleuropneumonia (CCPP) in Ethiopia. The objectives of the review were thus to provide a pooled prevalence estimate of CCPP in the country and asses the level of in between study variance among the available reports. Manual and electronic search was conducted between 8th of January and 25th of June 2015. A total of twelve published articles and one MSc thesis was retrieved from19 initially identified studies. Twenty five animal level datasets were extracted at regional level considering some hypothesized predictors. The retrieved data were summarized in a meta-analytical approach. Accordingly, the pooled prevalence estimate of CCPP was 25.7% (95% CI:20.9,31.0). The inverse variance square (I(2)) that explains the variation in effect size attributed to reports true heterogeneity was 95.7%.The sub-group analysis was also computed for assumed predictors including, age, sex, type of study population, production systems and regional states. Among these predictors, study population type revealed statistically significant difference (Paccounting for 22.87% of the explainable proportion of heterogeneity among the presumed predictors. Evidence on isolation and confirmation of Mycoplasma capricolum subspp. capripneumonie in the country was obtained from five regional states. In conclusion, it is recommended to further investigate facilities related with transportation and collection premises along with potential role of sheep in the epidemiology of CCPP. Finally, the review emphasizes the need for monitoring the ongoing CCPP control intervention and introduces amendments based on the findings. Besides more surveys are needed in some of the regions where no or few valid data was available. PMID:26943994

  4. Evaluation of Molecular Methods for Serotyping Shigella flexneri.

    Gentle, Amy; Ashton, Philip M; Dallman, Timothy J; Jenkins, Claire


    Shigella flexneri can be phenotypically serotyped using antisera raised to type-specific somatic antigens and group factor antigens and genotypically serotyped using PCR targeting O-antigen synthesis or modification genes. The aim of this study was to evaluate a real-time PCR for serotyping S. flexneri and to use whole-genome sequencing (WGS) to investigate the phenotypic and genotypic serotype identifications. Of the 244 cultures tested retrospectively, 226 (92.6%) had concordant results between phenotypic serotyping and PCR. Seventy of the 244 isolates (including 15 of the 18 isolates where a serotype-PCR mismatch was identified) were whole-genome sequenced, and the serotype was derived from the genome. Discrepant results between the phenotypic and genotypic tests were attributed to insertions/deletions or point mutations identified in O-antigen synthesis or modification genes, rendering them dysfunctional; inconclusive serotyping results due to nonspecific cross-reactions; or novel genotypes. Phylogenetic analysis of the WGS data indicated that the serotype, regardless of whether it was phenotypically or genotypically determined, was a weak predictor of phylogenetic relationships between strains of S. flexneri WGS data provided both genome-derived serotyping, thus supporting backward compatibility with historical data and facilitating data exchange in the community, and more robust and discriminatory typing at the single-nucleotide-polymorphism level. PMID:26984974

  5. Significance of CO2 donor on the production of succinic acid by Actinobacillus succinogenes ATCC 55618

    Zou Wei


    Full Text Available Abstract Background Succinic acid is a building-block chemical which could be used as the precursor of many industrial products. The dissolved CO2 concentration in the fermentation broth could strongly regulate the metabolic flux of carbon and the activity of phosphoenolpyruvate (PEP carboxykinase, which are the important committed steps for the biosynthesis of succinic acid by Actinobacillus succinogenes. Previous reports showed that succinic acid production could be promoted by regulating the supply of CO2 donor in the fermentation broth. Therefore, the effects of dissolved CO2 concentration and MgCO3 on the fermentation process should be investigated. In this article, we studied the impacts of gaseous CO2 partial pressure, dissolved CO2 concentration, and the addition amount of MgCO3 on succinic acid production by Actinobacillus succinogenes ATCC 55618. We also demonstrated that gaseous CO2 could be removed when MgCO3 was fully supplied. Results An effective CO2 quantitative mathematical model was developed to calculate the dissolved CO2 concentration in the fermentation broth. The highest succinic acid production of 61.92 g/L was obtained at 159.22 mM dissolved CO2 concentration, which was supplied by 40 g/L MgCO3 at the CO2 partial pressure of 101.33 kPa. When MgCO3 was used as the only CO2 donor, a maximal succinic acid production of 56.1 g/L was obtained, which was just decreased by 7.03% compared with that obtained under the supply of gaseous CO2 and MgCO3. Conclusions Besides the high dissolved CO2 concentration, the excessive addition of MgCO3 was beneficial to promote the succinic acid synthesis. This was the first report investigating the replaceable of gaseous CO2 in the fermentation of succinic acid. The results obtained in this study may be useful for reducing the cost of succinic acid fermentation process.

  6. Leptospira interrogans serotype hardjo in dairy cows

    Vidić Branka M.; Boboš Stanko F.


    Data on L. hardjo infection of dairy cows in the world pint out its important role in the occurrence of health and economic problem. L. interrogans serotype hardjo has been described as the cause of miscarriages, stillbirts, or the birhs of poorly vital calves, agalactia, mastitis, and low fertility in cows. Two L. hardjo genotypes have been identified in cows, namely, hardjopraitno and hardjobovis. Serological investigations have established a drastic increase in this leptospiral infection i...

  7. Possible involvement of protein kinase C in apoptotic cell death of macrophages infected with Actinobacillus actinomycetemcomitans.

    Nonaka, K; Ishisaki, A; Muro, M; Kato, S; Oido, M; Nakashima, K; Kowashi, Y; Nishihara, T


    We have previously reported the evidence for apoptosis in the mouse macrophage cell line J774.1 by the periodontopathic bacterium Actinobacillus actinomycetemcomitans. In this study, we examined the role of protein kinases in the induction of apoptosis in A. actinomycetemcomitans-infected J774.1 cells by the MTT assay, fluorescence microscopy and flow cytometric analysis. After J774.1 cells were precultured with protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA), J774.1 cells infected with A. actinomycetemcomitans showed the increased percentage of apoptotic cells. On the contrary, protein kinase A (PKA) activators, such as forskolin and dibutyryl cAMP, do not mimic the effect of PMA. PKC inhibitors, such as staurosporine, calphostin C, chelerythrine chloride, and H7 were found to suppress apoptotic cell death in J774.1 cells infected with A. actinomycetemcomitans. However, HA1004, known as PKA inhibitor, had no effect on apoptosis in infected macrophages. The results presented here suggest that the signals through PKC may play crucial roles in the modulation of apoptosis in macrophages infected with A. actinomycetemcomitans. PMID:9503618

  8. Optimization of succinic acid fermentation with Actinobacillus succinogenes by response surface methodology (RSM)

    Yun-jian ZHANG; Qiang LI; Yu-xiu ZHANG; Dan WANG; Jian-min XING


    Succinic acid is considered as an important platform chemical.Succinic acid fermentation with Actinobacillus succinogenes strain BE-1 was optimized by central composite design (CCD) using a response surface methodology (RSM).The optimized production of succinic acid was predicted and the interactive effects between glucose,yeast extract,and magnesium carbonate were investigated.As a result,a model for predicting the concentration of succinic acid production was developed.The accuracy of the model was confirmed by the analysis of variance (ANOVA),and the validity was further proved by verification experiments showing that percentage errors between actual and predicted values varied from 3.02% to 6.38%.In addition,it was observed that the interactive effect between yeast extract and magnesium carbonate was statistically significant.In conclusion,RSM is an effective and useful method for optimizing the medium components and investigating the interactive effects,and can provide valuable information for succinic acid scale-up fermentation using A.succinogenes strain BE-1.

  9. A complete industrial system for economical succinic acid production by Actinobacillus succinogenes.

    Li, Jian; Zheng, Xiao-Yu; Fang, Xiao-Jiang; Liu, Shu-Wen; Chen, Ke-Quan; Jiang, Min; Wei, Ping; Ouyang, Ping-Kai


    An industrial fermentation system using lignocellulosic hydrolysate, waste yeast hydrolysate, and mixed alkali to achieve high-yield, economical succinic acid production by Actinobacillus succinogenes was developed. Lignocellulosic hydrolysate and waste yeast hydrolysate were used efficiently as carbon sources and nitrogen source instead of the expensive glucose and yeast extract. Moreover, as a novel method for regulating pH mixed alkalis (Mg(OH)(2) and NaOH) were first used to replace the expensive MgCO(3) for succinic acid production. Using the three aforementioned substitutions, the total fermentation cost decreased by 55.9%, and 56.4 g/L succinic acid with yield of 0.73 g/g was obtained, which are almost the same production level as fermentation with glucose, yeast extract and MgCO(3). Therefore, the cheap carbon and nitrogen sources, as well as the mixed alkaline neutralize could be efficiently used instead of expensive composition for industrial succinic acid production. PMID:21470857

  10. Detection of highly and minimally leukotoxic Actinobacillus actinomycetemcomitans strains in patients with periodontal disease

    Cortelli Sheila Cavalca


    Full Text Available This study examined the prevalence of highly and minimally leukotoxic Actinobacillus actinomycetemcomitans in patients with periodontal disease. Pooled subgingival plaque samples from 136 patients with some form of periodontal disease were examined. Subjects were between 14 and 76 years of age. Clinical examinations included periodontal pocket depth (PD, plaque index (PI and bleeding index (BI. The obtained plaque samples were examined for the presence of highly or minimally leukotoxic A. actinomycetemcomitans strains by the polymerase chain reaction (PCR. Chi-square and logistic regression were performed to evaluate the results. Forty-seven subjects were diagnosed with gingivitis, 70 with chronic periodontitis and 19 with aggressive periodontitis. According to chi-square there was no significant correlation detected between PD (chi2 = 0.73, PI (chi2 = 0.35, BI (chi2 = 0.09 and the presence of the highly leukotoxic A. actinomycetemcomitans. The highly leukotoxic A. actinomycetemcomitans strains were correlated with subjects that were 28 years of age and younger (chi2 = 7.41. There was a significant correlation between highly leukotoxic A. actinomycetemcomitans and aggressive periodontitis (chi2 = 22.06. This study of a Brazilian cohort confirms the strong association between highly leukotoxic A. actinomycetemcomitans strains and the presence of aggressive periodontitis.

  11. Rapid detection of Actinobacillus actinomycetemcomitans, Prevotella intermedia and Porphyromona gingivalis by multiplex PCR.

    García, L; Tercero, J C; Legido, B; Ramos, J A; Alemany, J; Sanz, M


    The identification of specific periodontal pathogens by conventional methods, mainly anaerobic cultivation, is difficult, time consuming and even sometimes unreliable. Therefore, a multiplex PCR method for simultaneous detection of Actinobacillus actinomycetemcomitans (A.a.), Porphyromona gingivalis (P.g.) and Prevotella intermedia (P.i.) was developed for rapid and easy identification of these specific bacterial pathogens in subgingival plaque samples. In this paper, there is a detailed description of the oligonucleotide primer selection, DNA extraction and PCR conditions and the sequencing of the amplified products. The locus chosen to be amplified is a highly variable region in the 16S ribosomal DNA. For the development of this technique ATCC cultures and pure cultures from subgingival plaque samples taken from periodontitis patients were used. As an internal positive control a recombinant plasmid was developed. This simple DNA extraction procedure and the DNA amplification and visualization of the amplified product permits the detection of the bacteria in a working day. Thus, this multiplex PCR method is a rapid and effective detection method for specific periodontal pathogens. PMID:9524322

  12. Differential transcription of virulence genes in Aggregatibacter actinomycetemcomitans serotypes

    Marcia Pinto Alves MAYER; Umeda, Josely Emiko; Priscila Larcher LONGO; Simionato, Maria Regina Lorenzetti


    Background: Aggregatibacter actinomycetemcomitans serotypes are clearly associated with periodontitis or health, which suggests distinct strategies for survival within the host.Objective: We investigated the transcription profile of virulence-associated genes in A. actinomycetemcomitans serotype b (JP2 and SUNY 465) strains associated with disease and serotype a (ATCC 29523) strain associated with health. Design: Bacteria were co-cultured with immortalized gingival epithelial cells (OBA-9). T...

  13. Serotypes and penicillin susceptibility of pneumococci isolated from blood.

    Lauer, B A; Reller, L B


    To learn the prevalence of penicillin-resistant pneumococci and the distribution of serotypes in proved pneumococcal infections, we studied 98 pneumococci recovered from blood over a 4-year period. Penicillin susceptibility was determined by the agar dilution method. Serotyping was done by the capsular swelling (quellung) test. Only one strain showed diminished susceptibility to penicillin (minimal inhibitory concentration, 0.12 micrograms/ml). Twenty-three different serotypes were identified...

  14. The use of oligonucleotide probes for meningococcal serotype characterization

    SACCHI Claudio Tavares


    Full Text Available In the present study we examine the potential use of oligonucleotide probes to characterize Neisseria meningitidis serotypes without the use of monoclonal antibodies (MAbs. Antigenic diversity on PorB protein forms the bases of serotyping method. However, the current panel of MAbs underestimated, by at least 50% the PorB variability, presumably because reagents for several PorB variable regions (VRs are lacking, or because a number of VR variants are not recognized by serotype-defining MAbs12. We analyzed the use of oligonucleotide probes to characterize serotype 10 and serotype 19 of N. meningitidis. The porB gene sequence for the prototype strain of serotype 10 was determined, aligned with 7 other porB sequences from different serotypes, and analysis of individual VRs were performed. The results of DNA probes 21U (VR1-A and 615U (VR3-B used against 72 N. meningitidis strains confirm that VR1 type A and VR3 type B encode epitopes for serotype-defined MAbs 19 and 10, respectively. The use of probes for characterizing serotypes possible can type 100% of the PorB VR diversity. It is a simple and rapid method specially useful for analysis of large number of samples.

  15. Pneumococcal Serotype 19F Conjugate Vaccine Induces Cross-Protective Immunity to Serotype 19A in a Murine Pneumococcal Pneumonia Model

    Jakobsen, Håvard; Sigurdsson, Viktor D.; Sigurdardottir, Sigurveig; Schulz, Dominique; Jonsdottir, Ingileif


    Immunization with a pneumococcal conjugate vaccine (PNC) containing serotype 19F induces cross-reactive antibodies to 19A in mice and human infants. Active immunization with PNC and passive immunization with serum samples from infants vaccinated with PNC containing serotype 19F, but not serotype 19A, protected against lung infection caused by both serotypes in a murine model.

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

    Sun Qi; Rodriguez-Rivera Lorraine D; Orsi Renato H; Soyer Yeşim; Wiedmann Martin


    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 ...

  17. An outbreak of contagious bovine pleuropneumonia (CBPP) and post outbreak sero-surveillance in Eritrea

    At the end of 2002 contagious bovine pleuropneumonia (CBPP) was never reported in Eritrea for the last 20 to 30 years. A major incursion of CBPP was reported from illegally smuggled Raya Azebo type cattle from Ethiopia, introduced through the Southern Red Sea administrative region of Eritrea (the Afar area bordering Ethiopia). The first case was detected in the Asmara abattoir during post mortem inspection. Characteristic CBPP clinical signs and post-mortem lesions were observed in affected herd during the outbreak of the disease. Significant number of in-contact indigenous cattle was infected by the disease. Samples were first sent to OIE reference laboratory CIRADEMVT in France for confirmation. The disease was confirmed by PCR on 15 January 2003.Samples of Pleural fluids and lung tissues were collected from CBPP infected animals and tested using the polymerase chain reaction (PCR) technique at the National Veterinary Laboratory. The affected cattle herds were held in an isolation pen and eventually destroyed following confirmation of the disease. Following this incident, surveillance of CBPP was stepped-up and a total of 4,695 serum samples were collected from 313 sampling units or villages and submitted to the National Veterinary Laboratory for testing in 2004 one year after the outbreak of the disease as part of the epidemiosurveillance. Serum samples collected for sero-surveillance were tested for CBPP antibodies by the use of the monoclonal antibody specific for Mycoplasma mycoides subspecies mycoides sc as described in the competitive ELISA kit manual from CIRAD-EMVT, France. During this post outbreak sero-surveillance sampling season, all age group animals were eligible for sampling. Out of the total sampled animals 67 (1.43%) were positive for Mycoplasma mycoides subsp. mycoides sc antibodies using the cELISA test. This result suggested that very low lateral infectious agent transmission could have taken place. Vaccination as a control measure had never

  18. Bacterial Load of Pneumococcal Serotypes Correlates with Their Prevalence and Multiple Serotypes Is Associated with Acute Respiratory Infections among Children Less Than 5 Years of Age

    Bhim Gopal Dhoubhadel; Michio Yasunami; Hien Anh Thi Nguyen; Motoi Suzuki; Thu Huong Vu; Ai Thi Thuy Nguyen; Duc Anh Dang; Lay-Myint Yoshida; Koya Ariyoshi


    BACKGROUND: Among pneumococcal serotypes, some serotypes are more prevalent in the nasopharynx than others; determining factors for higher prevalence remain to be fully explored. As non-vaccine serotypes have emerged after the introduction of 7-valent conjugate vaccines, study of serotype specific epidemiology is in need. When two or more serotypes co-colonize, they evolve rapidly to defend host's immune responses; however, a clear association of co-colonization with a clinical outcome is lac...

  19. Sero-positivity and associated risk factors for contagious bovine pleuropneumonia under two cattle production systems in North Central Nigeria.

    Alhaji, Nma Bida; Babalobi, Olutayo Olajide


    A cross-sectional survey of 765 cattle in 125 nomadic and 375 cattle in 125 sedentary herds was conducted to investigate prevalence and risk factors for contagious bovine pleuropneumonia (CBPP) in the two production systems of Niger State in North Central Nigeria, between January and August 2013. Data on herd characteristics were collected using structured questionnaires administered on herd owners. Serological analysis was conducted using competitive enzyme linked immunosorbent assay (c-ELISA) test. Descriptive, univariate, and multivariate statistical analyses were conducted with OpenEpi version 2.3.1 software. Statistical significance was held at P trekking during migrations (P = 0.0009). This study had shown the burden of CBPP in the two production systems. Sero-diagnosis and risk factor identification should be institutionalized as elements of epidemio-surveillance and control strategies for CBPP, especially in resource-poor pastoralists' settlements in Nigeria. PMID:26563271

  20. An abattoir survey of contagious bovine pleuropneumonia lesions in slaughtered cattle in selected districts in Northern Tanzania

    Emmanuel; Swai; Isidory; Mwezimpya; Edward; Ulicky; Adam; Mbise; Winford; Moshy


    Objective:To establish and estimate incidence of contagious bovine pleuropneumonia(CBPP),using abattoir survey as a diagnostic tool in slaughtered cattle in Northern Tanzania.Methods:A total of 4460 cattle were slaughtered in five abattoirs in 3 northern zone regions(Arusha,Kilimanjaro and Tanga)during the period of January to May 2004.They were examined ante-mortem for‘pneumonia signs’,and‘characteristic contagious bovine pleuropneumonia(CBPP)lung lesions’.Results:Forty-one(0.91%)of the slaughtered cattle,the majority of which were Tanzania short horn zebu,had gross lung lesions suggestive of CBPP.The prevalence of lesions was significantly(P<0.05)higher in Karatu abattoir compared to others.No animal was detected to have lesion in Bomang’ombe abattoir.The most observed pneumonic signs included labored breathing(90%),dry cough(57%)and mucopurulent nasal discharge(47%).The gross characteristic CBPP pathological lesion,frequently encountered was left lung lesion(47%),pinkish lung(71%)and pleural adhesion(98%).Epidemiological reports show that the CBPP reported outbreaks increased from 19 in 2002,65 in 2003 and 18 in 2004(January-March).The corresponding number of reported deaths increased from 137 in 2002,269 in 2003 and 77 in 2004(January-March).Conclusions:It’s concluded from this study that CBPP is a problem in spite of the extensive awareness and vaccination campaigns.Nevertheless,a continued surveillance programme including routine checks of all cattle carcasses at the abattoir and subsequent epidemiological investigation of suspected cases are recommended.

  1. Probability of identifying different salmonella serotypes in poultry samples

    Recent work has called attention to the unequal competitive abilities of different Salmonella serotypes in standard broth culture and plating media. Such serotypes include Enteritidis and Typhimurium that are specifically targeted in some regulatory and certification programs because they cause a l...

  2. Antibiotic Susceptibilities and Serotyping of Clinical Streptococcus Agalactiae Isolates

    Altay Atalay


    Full Text Available Objective: Streptococcus agalactiae (Group B streptococci, GBS are frequently responsible for sepsis and meningitis seen in the early weeks of life. GBS may cause perinatal infection and premature birth in pregnant women. The aim of this study was to serotype GBS strains isolated from clinical samples and evaluate their serotype distribution according to their susceptibilities to antibiotics and isolation sites. Material and Methods: One hundred thirty one S. agalactiae strains isolated from the clinical samples were included in the study. Of the strains, 99 were isolated from urine, 20 from soft tissue, 10 from blood and 2 from vaginal swab. Penicillin G and ceftriaxone susceptibilities of GBS were determined by the agar dilution method. Susceptibilities to erythromycin, clindamycin, vancomycin and tetracycline were determined by the Kirby-Bauer method according to CLSI criteria. Serotyping was performed using the latex aglutination method using specific antisera (Ia, Ib, II-VIII. Results: While in 131 GBS strains, serotypes VII and VIII were not detected, the most frequently isolated serotypes were types Ia (36%, III (30.5% and II (13% respectively. Serotype Ia was the most frequently seen serotype in all samples. All GBS isolates were susceptible to penicilin G, ceftriaxone and vancomycin. Among the strains, tetracycline, erythromycin and clindamycin resistance rates were determined as 90%, 14.5%, and 13% respectively. Conclusion: Penicillin is still the first choice of treatment for the infections with all serotypes of S. agalactiae in Turkey.

  3. Nitric oxide production by murine spleen cells stimulated with lipopolysaccharide from Actinobacillus actinomycetemcomitans.

    Sosroseno, Wihaskoro; Herminajeng, Endang; Susilowati, Heni; Budiarti, Sri


    The aim of this study was to determine whether Actinobacillus actinomycetemcomitans lipopolysaccharide (LPS-A. actinomycetemcomitans) could induce murine spleen cells to produce nitric oxide (NO). Spleen cells derived from Balb/c mice were stimulated with LPS-A. actinomycetemcomitans or LPS from Escherichia coli for 4 days. The effects of N(G)-monomethyl-L-arginine (NMMA), polymyxin B, and cytokines (IFN-gamma and IL-4) on the production of NO were also assessed. The NO production from the carrageenan-treated spleen cells stimulated with LPS-A. actinomycetemcomitans or both LPS-A. actinomycetemcomitans and IFN-gamma was determined. The carrageenan-treated mice were transferred with splenic macrophages and the NO production was assessed from the spleen cells stimulated with LPS-A. actinomycetemcomitans or LPS-A. actinomycetemcomitans and IFN-gamma. The results showed that NO production was detectable in the cultures of spleen cells stimulated with LPS-A. actinomycetemcomitans in a dose-dependent fashion, but was lower than in the cells stimulated with LPS from E. coli. The NO production was blocked by NMMA and polymyxin B. IFN-gamma up-regulated but IL-4 suppressed the production of NO by the spleen cells stimulated with LPS-A. actinomycetemcomitans. The carrageenan-treated spleen cells failed to produce NO after stimulation with LPS-A. actinomycetemcomitans or both LPS-A. actinomycetemcomitans and IFN-gamma. Adoptive transfer of splenic macrophages to the carrageenan-treated mice could restore the ability of the spleen cells to produce NO. The results of the present study suggest that LPS-A. actinomycetemcomitans under the regulatory control of cytokines induces murine spleen cells to produce NO and that splenic macrophages are the cellular source of the NO production. Therefore, these results may support the view that NO production by LPS-A. actinomycetemcomitans-stimulated macrophages may play a role in the course of periodontal diseases. PMID:16887678

  4. Salmonella serotypes in reptiles and humans, French Guiana.

    Gay, Noellie; Le Hello, Simon; Weill, François-Xavier; de Thoisy, Benoit; Berger, Franck


    In French Guiana, a French overseas territory located in the South American northern coast, nearly 50% of Salmonella serotypes isolated from human infections belong to serotypes rarely encountered in metropolitan France. A reptilian source of contamination has been investigated. Between April and June 2011, in the area around Cayenne, 151 reptiles were collected: 38 lizards, 37 snakes, 32 turtles, 23 green iguanas and 21 caimans. Cloacal swab samples were collected and cultured. Isolated Salmonella strains were identified biochemically and serotyped. The overall carriage frequency of carriage was 23.2% (95% confidence interval: 16.7-30.4) with 23 serotyped strains. The frequency of Salmonella carriage was significantly higher for wild reptiles. Near two-thirds of the Salmonella serotypes isolated from reptiles were also isolated from patients in French Guiana. Our results highlight the risk associated with the handling and consumption of reptiles and their role in the spread of Salmonella in the environment. PMID:24560590

  5. Serotypes of Streptococcus pneumoniae causing major pneumococcal infections

    Yu. V. Lobzin


    Full Text Available First in Russia prospective non-interventional hospital-based study on Streptococcus pneumoniae serotypes causing meningitis and acute otitis media (AOM in children and community-acquired pneumonia (CAP in children and adults, as well as serotype coverage by pneumococcal conjugate vaccines (PCV’s of different composition has been conducted. Serotypes 19F, 14 and serogroup 6 are the leading in meningitis; serotype coverage is 70,6% for PCV7, and 76,5% – for PCV10 and PCV13. Among S. pneumoniae serotypes causing AOM 19F, 3, 23F and serogroup 6 have been the most prevalent in Saint Petersburg. PCV7 and PCV10 provide equal serotypes coverage in AOM – 63,2% among children 0–2 years old, and 32,5% among children 5–17 years old. PCV13 covers up to 79% of serotypes in infants. In CAP PCV7 and PCV10 provide 57,1% serotype coverage in children and 56,1% – in adults. Serotype coverage in CAP for PCV13 has been 14,3% and 34,5% higher for children and adults, correspondingly. Obtained data supports PCV inclusion in children immunization program in Saint Petersburg, whereas PCV13 provides the broadest serotype coverage. In the course PCV’s implementation continued pneumococcal infection surveillance is advisable.

  6. Contagious bovine pleuropneumonia (CBPP) in the Maasai ecosystem of south-western Kenya: Evaluation of seroprevalence, risk factors and vaccine safety and efficacy.

    Mtui-Malamsha, Niwael Jesse


    Contagious bovine pleuropneumonia (CBPP) is a bovine bacterial disease of major economic importance in sub-Saharan Africa. Vaccination has been recommended to control the disease in endemic areas such as the Maasai ecosystems of Kenya and Tanzania; however, the currently used live attenuated vaccine has been reported to have poor vaccine safety and efficacy. To compare standard (current) and an improved (buffered) version of the live CBPP-vaccine, several epidemiological studie...

  7. Serotype-specific and serotype-independent strategies for pre-harvest control of foodborne Salmonella in poultry.

    Of more than 2500 identified Salmonella serotypes, only a small proportion are common in poultry flocks. However, there is an epidemiologically important connection between poultry products and human infections, as many of the serotypes that are most prevalent in humans (such as S. Typhimurium and S...

  8. Selective and genetic constraints on pneumococcal serotype switching.

    Nicholas J Croucher


    Full Text Available Streptococcus pneumoniae isolates typically express one of over 90 immunologically distinguishable polysaccharide capsules (serotypes, which can be classified into "serogroups" based on cross-reactivity with certain antibodies. Pneumococci can alter their serotype through recombinations affecting the capsule polysaccharide synthesis (cps locus. Twenty such "serotype switching" events were fully characterised using a collection of 616 whole genome sequences from systematic surveys of pneumococcal carriage. Eleven of these were within-serogroup switches, representing a highly significant (p < 0.0001 enrichment based on the observed serotype distribution. Whereas the recombinations resulting in between-serogroup switches all spanned the entire cps locus, some of those that caused within-serogroup switches did not. However, higher rates of within-serogroup switching could not be fully explained by either more frequent, shorter recombinations, nor by genetic linkage to genes involved in β-lactam resistance. This suggested the observed pattern was a consequence of selection for preserving serogroup. Phenotyping of strains constructed to express different serotypes in common genetic backgrounds was used to test whether genotypes were physiologically adapted to particular serogroups. These data were consistent with epistatic interactions between the cps locus and the rest of the genome that were specific to serotype, but not serogroup, meaning they were unlikely to account for the observed distribution of capsule types. Exclusion of these genetic and physiological hypotheses suggested future work should focus on alternative mechanisms, such as host immunity spanning multiple serotypes within the same serogroup, which might explain the observed pattern.

  9. Reinfections and rotavirus serotypes in Belém, Brazil

    Alexandre da C. Linhares


    Full Text Available Repeated infections involving different rotavirus serotypes were detected in four children living in Belém, who were followed up since birth to three years of age. In one child (Reg. 23.983 three successive symptomatic infections (one of them associated with serotype 2 were noted: the first, at four months of age, the second at 20 months and the third at 27 months. Another child had two subsequent infections, the first one by rotavirus serotype 1, and the second by a not identified rotavirus serotype. In this case two episodes of rotavirus-related diarrhoea were recorded, occurring eight months apart. Apparent infections were detected on two occasions involving the same child (Reg. 24.004, the first being associated with serotype 1, and the second with serotype 2. The fourth child (Reg. 24.097 had two successive infections by not determined rotavirus serotypes, without clinical manifestations; the first occurred at 24 months and the second at 28 months.

  10. Draft Genome Sequences of Streptococcus agalactiae Serotype Ia and III Isolates from Tilapia Farms in Thailand

    Areechon, Nontawith; Kannika, Korntip; Hirono, Ikuo; Kondo, Hidehiro; Unajak, Sasimanas


    Streptococcus agalactiae serotypes Ia and III were isolated from infected tilapia in cage and pond culture farms in Thailand during 2012 to 2014, in which pathogenicity analysis demonstrated that serotype III showed higher virulence than serotype Ia. Here, we report the draft genome sequencing of piscine S. agalactiae serotypes Ia and III.

  11. Dengue virus serotype in Aceh Province



    Full Text Available WHO estimated 50 million dengue infections happen every year in the world. In Indonesia, there were 90,245 DHF cases on 2012 with 816 deaths. In the Province of Aceh, 2,269 cases happened in the same year. This study aimed to identify dengue virus serotype in Aceh. Sampling was done in Kota Banda Aceh Hospital, Kota Lhokseumawe Hospital, Kabupaten Aceh Tamiang Hospital, Kabupaten Aceh Barat Hospital, and Kabupaten Simeulue Hospital between May to December 2012. This was a clinical laboratory research with observation design using cross sectional approach. Research’s population was sample from patients with dengue clinical symptom. Using purposive sampling technique, we have collected 100 samples from the five hospitals (20 samples from each hospital. From RT-PCR, we found 16 positive samples (9 samples were DENV-4, 3 samples were DENV-1, 2 samples were DENV-2, and 2 samples were DENV-3.

  12. The role of Actinobacillus actinomycetemcomitans fimbrial adhesin on MMP-8 activity in aggressive periodontitis pathogenesis

    Rini Devijanti Ridwan


    Full Text Available Background: Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans is Gram negative and a major bacterial agent associated with aggressive periodontitis in young adult, this bacteria was an important factor in pathogenesis of aggressive periodontitis. A. actinomycetemcomitans possesses fimbriae with an adhesin protein that was the first bacterial molecules to make physical contact with host. Purpose: The objective of this research was to analyzed the influence of A. actinomycetemcomitans fimbrial adhesin protein induction on MMP-8 activity. Methods: The research was an experimental laboratory study, the step in this study were isolation and identification A. actinomycetemcomitans, characterize A. actinomycetemcomitans adhesin and study the role of A. actinomycetemcomitans adhesin in Wistar rats. Results: The result of this research on the role of adhesin in Wistar rats after analysis with Analysis of Variance (ANOVA showed significant differences in the control group with group induction with A. actinomycetemcomitans, A. actinomycetemcomitans plus adhesin and adhesin. MMP-8 activity increased with induction A. actinomycetemcomitans and 24 kDa A. actinomycetemcomitans adhesin. This fimbrial adhesin protein showed that A. actinomycetemcomitans has the ability to adhesion, colonization and invasion for host in aggressive periodontitis pathogenesis. Conclusion: A. actinomycetemcomitans fimbrial adhesin protein induction increasing MMP-8 activity for aggressive periodontitis pathogenesis.Latar belakang: A. actinomycetemcomitans merupakan salah satu bakteri Gram negatif yang terkait dengan periodontitis agresif yang menyerang penderita usia muda dan merupakan faktor penting dalam patogenesis periodontitis agresif. A. actimycetemcomitans mempunyai fimbriae dengan protein adhesin yang merupakan molekul pertama dari bakteri untuk melakukan kontak fisik dengan host. Tujuan: Tujuan penelitian ini adalah menganalisis pengaruh induksi adhesin A

  13. Leptospira interrogans serotype hardjo in dairy cows

    Vidić Branka M.


    Full Text Available Data on L. hardjo infection of dairy cows in the world pint out its important role in the occurrence of health and economic problem. L. interrogans serotype hardjo has been described as the cause of miscarriages, stillbirts, or the birhs of poorly vital calves, agalactia, mastitis, and low fertility in cows. Two L. hardjo genotypes have been identified in cows, namely, hardjopraitno and hardjobovis. Serological investigations have established a drastic increase in this leptospiral infection in cows. L. hardjo has become adapted to cattle as the primary host, so that an infection is maintained in herds and becomes deeply rooted because of the permanent presence of the source of infection. It was believed that sheep were accidental hosts, but the latest research suggest that they are yet another, transitory, host for maintining this leptospira serotype. L. hardjo is also important from the aspect of human health, especially of persons who are professionally exposed to this infection. L. hardjo infection is detected using serological tests and by proving the presence of leptospira. The medicine of choice in the therapy of leptospiral infections is streptomycin (DSM. Therapy using oxytetracyclines for clinical mastitis was also proven effective. Treatment is most successful in the early stage of the disease. A single dose of streptomycin administered in infected herds reduces the duration period of leptospira excretion through urine, thus preventing the spread of infection thorugh contaminated urine. The basic components of the plan to contain leptospira are the following: serological investigations, sanitary-higiene measures, the elimination of animals which excrete leptospira through urine, therapy, vaccination, quarantine.

  14. SALMATcor: microagglutination for Salmonella flagella serotyping.

    Duarte Martínez, Francisco; Sánchez-Salazar, Luz Marina; Acuña-Calvo, María Teresa; Bolaños-Acuña, Hilda María; Dittel-Dittel, Isis; Campos-Chacón, Elena


    Salmonella is a complex bacterial group with more than 2400 serovars widely distributed in nature; they are considered zoonotic because they can infect a variety of animals and be transmitted to humans. Usually, they cause alimentary acquired diseases such as gastroenteritis, typhoid fever, and others that can lead to severe complications and death. Serotyping is useful to differentiate among Salmonella, because it shows an important correlation with their clinical and epidemiological patterns; consequently, it is of high value for public health, animal health, agriculture, and industry. To characterize all known Kauffmann-White Salmonella serovars, over 250 antisera are required. Due to this and to high prices antisera, many laboratories worldwide have limitations in establishing Salmonella surveillance. Therefore, we developed and validated a Salmonella flagella microagglutination test (SALMATcor) that significantly reduces laboratory requirements of antisera. SALMATcor is based on scaling down, by fivefold, the antigen:antiserum volumes actually required for the reference method: flagella standard tube agglutination technique (STAT). Antigen preparation, temperatures, and incubation periods remained as established for STAT. The SALMATcor was validated according to ISO/DIS 16140:1999 protocol, which included 1187 comparisons of flagella determinations conducted by SALMATcor and STAT, on 141 Salmonella isolates of 12 common serotypes and the use of antiserum recommended for STAT. SALMATcor concordance was excellent (Cohen's kappa index 0.9982), obtaining relative accuracy >99.9% and relative specificity >99.9%. Additionally, SALMATcor has been used by CNRB-INCIENSA since 2004 to respond to all 40 Salmonella proficiency testing strains, provided by World Health Organization-Global Salmonella Surveillance Network, obtaining 100% concordance on serovar identification. On the basis of the results achieved with SALMATcor and considering that it also significantly

  15. Disease risks associated with free-ranging wild boar in Saskatchewan.

    McGregor, Glenna F; Gottschalk, Marcelo; Godson, Dale L; Wilkins, Wendy; Bollinger, Trent K


    This study investigated the disease status of Saskatchewan's feral wild boar population. Whole carcasses, tissue samples, and/or serum from 81 hunter-killed boars from Saskatchewan were submitted to the Canadian Wildlife Health Cooperative (CWHC) between 2009 and 2014. Serological tests were negative for PRRS, H1N1, and H3N2 swine influenza, PCV-2, and TGE/PRCV in 22/22 boars and for Toxoplasma gondii and Mycoplasma hyopneumoniae in 20/20 boars. Of 20 boars whose sera were tested 20 were positive for Actinobacillus pleuropneumoniae, with 7 positive for, among other strains, serotype 14; 16 were positive for Lawsonia intracellularis, 1 was positive and 6 were suspicious for Salmonella spp. Polymerase chain reaction tests were negative for PRRS and PCV2 in 58/58 boars and positive for Torque teno virus in 1/8 boars. Digestion assays were negative for Trichinella spp. in 22/22 boars. The high seroprevalence of A. pleuropneumoniae serotype 14 is noteworthy as this serotype has not been previously reported in North America. PMID:26246630

  16. Analysis of Salmonella enterica serotype-host specificity in calves: Avirulence of S-enterica serotype gallinarum correlates with bacterial dissemination from mesenteric lymph nodes and persistence in vivo

    Paulin, S M; Watson, P. R.; Benmore, A R; M.P. Stevens; Jones, P W; Villarreal-Ramos, B.; Wallis, T S


    Host and bacterial factors that determine whether Salmonella serotypes remain restricted to the gastrointestinal tract or penetrate beyond the mucosa and cause systemic disease remain largely undefined. Here, factors influencing Salmonella host specificity in calves were assessed by characterizing the pathogenesis of different serotypes. Salmonella enterica serotype Dublin was highly virulent intravenously, whereas S. enterica serotype Choleraesuis was moderately virulent. Both serotypes were...

  17. Salmonella enterica serotype dublin bacteraemia mimicking enteric fever

    Dias M; Antony B; Pinto H; Rekha B.


    Salmonella enterica serotype Dublin, a bovine adapted serotype, is the commonest cause of salmonellosis in cattle. Salmonellosis in animals always presents a potential zoonotic threat. Infected cattles serves as a source of infection to humans. We present here Salmonella Dublin Bacteraemia in an elderly patient, with all the clinical details, due to the rarity of its occurrence. He was treated successfully with ciprofloxacin and his follow up period was uneventful.

  18. Salmonella enterica serotype dublin bacteraemia mimicking enteric fever

    Dias M


    Full Text Available Salmonella enterica serotype Dublin, a bovine adapted serotype, is the commonest cause of salmonellosis in cattle. Salmonellosis in animals always presents a potential zoonotic threat. Infected cattles serves as a source of infection to humans. We present here Salmonella Dublin Bacteraemia in an elderly patient, with all the clinical details, due to the rarity of its occurrence. He was treated successfully with ciprofloxacin and his follow up period was uneventful.

  19. Recombinant Actinobacillus actinomycetemcomitans Cytolethal Distending Toxin Proteins Are Required To Interact To Inhibit Human Cell Cycle Progression and To Stimulate Human Leukocyte Cytokine Synthesis

    Akifusa, Sumio; Poole, Stephen; Lewthwaite, Jo; Henderson, Brian; Nair, Sean P


    It has recently been discovered that Actinobacillus actinomycetemcomitans, an oral bacterium causing periodontitis, produces cytolethal distending toxin (CDT), a cell cycle-modulating toxin that has three protein subunits: CdtA, CdtB, and CdtC. In this study, we have cloned and expressed each toxin gene from A. actinomycetemcomitans in Escherichia coli and purified the recombinant Cdt proteins to homogeneity. Individual Cdt proteins failed to induce cell cycle arrest of the human epithelial c...

  20. Significance of CO2 donor on the production of succinic acid by Actinobacillus succinogenes ATCC 55618

    Zou Wei; Zhu Li-Wen; Li Hong-Mei; Tang Ya-Jie


    Abstract Background Succinic acid is a building-block chemical which could be used as the precursor of many industrial products. The dissolved CO2 concentration in the fermentation broth could strongly regulate the metabolic flux of carbon and the activity of phosphoenolpyruvate (PEP) carboxykinase, which are the important committed steps for the biosynthesis of succinic acid by Actinobacillus succinogenes. Previous reports showed that succinic acid production could be promoted by regulating ...

  1. Mouse interleukin-1 receptor antagonist induced by Actinobacillus actinomycetemcomitans lipopolysaccharide blocks the effects of interleukin-1 on bone resorption and osteoclast-like cell formation.

    Nishihara, T; Ohsaki, Y.; Ueda, N; Saito, N; Mundy, G R


    We have reported that P388D1 cell line murine macrophages stimulated with lipopolysaccharide (LPS) from Actinobacillus actinomycetemcomitans release interleukin-1 (IL-1) inhibitor. The IL-1 inhibitor was purified from conditioned media of P388D1 cells stimulated with A. actinomycetemcomitans LPS for 72 h to homogeneity by a four-step procedure: acetic acid extraction from conditioned media; Bio-Gel P-60 gel filtration chromatography; DEAE-Sepharose CL-6B column chromatography; and reverse-pha...

  2. Staphylococcus aureus Strains That Express Serotype 5 or Serotype 8 Capsular Polysaccharides Differ in Virulence

    Watts, Andrew; Ke, Danbing; Wang, Qun; Pillay, Anil; Nicholson-Weller, Anne; Lee, Jean C.


    Most isolates of Staphylococcus aureus produce a serotype 5 (CP5) or 8 (CP8) capsular polysaccharide. To investigate whether CP5 and CP8 differ in their biological properties, we created isogenic mutants of S. aureus Reynolds that expressed CP5, CP8, or no capsule. Biochemical analyses of CP5 and CP8 purified from the isogenic S. aureus strains were consistent with published structures. The degree of O acetylation of each polysaccharide was similar, but CP5 showed a greater degree of N acetyl...

  3. Sero-prevalence of Contagious Bovine Pleuropneumonia (CBPP) in bulls originated from Borena pastoral area of Southern Ethiopia.

    Alemayehu, Gezahegn; Leta, Samson; Hailu, Berhanu


    Contagious Bovine Pleuropneumonia (CBPP) is a highly infectious cattle disease, which is widespread in pastoral areas of Africa, and it imposes a major problem on Ethiopian livestock export market. Cross-sectional study was conducted in 2011 on bulls originated from Borena pastoral area to determine seroprevalence of CBPP. Forty batches of bulls containing 38,187 Borana bulls were tested using c-ELISA. Of the total 40 batches tested for the presence of antibodies, 25 (62.5 %) of them contained at least one seropositive bull. From the total of 38,187 bulls tested, 150 (0.4 %) bulls were positive. The number of seropositive animals increases as the herd size increases (P 1000, and the difference was found statistically significant (P opportunity to livestock producers and live animal and meat exporters by improving the demand of Ethiopian livestock on international market. Regular reintroduction of infected cattle from neighboring countries or herds where the disease remains endemic may change the disease dynamics again. Therefore, mass blanket vaccinations coupled with prompt diagnosis, isolation and stamping out of the outbreaks, intensive surveillance, followed by strict cattle movement control should be implemented by concerned parties. PMID:25863957

  4. Prevalence of contagious caprine pleuro-pneumonia in pastoral flocks of goats in the Rift Valley region of Kenya.

    Kipronoh, Alexander Kipruto; Ombui, Jackson Nyarongi; Kiara, Henry Kimathi; Binepal, Yatinder Singh; Gitonga, Eric; Wesonga, Hezron Okwako


    A cross-sectional survey was conducted between the months of March 2014 and March 2015 to determine the prevalence of contagious caprine pleuropneumonia in goat populations in pastoral flocks in three sub-counties of the Rift Valley region. A total of 432 serum samples were collected from goats from 54 flocks and tested for the presence of antibodies against mycoplasma capricolum subspecies capripneumoniae (mccp) using monoclonal antibody-based competitive enzyme-linked immuno-sorbent assay. Sero-prevalence recorded for Turkana West was 63.9%, Kajiado Central was 48.6%, while Pokot East was 29.2% which was statistically significant (χ2 = 34.997; P = 0.000) in the study sites. The results of this study confirmed that CCPP is widespread and endemic in the pastoral production systems studied in the Rift Valley region. The results confirmed that regions sharing international boundaries are at a higher risk of CCPP hence the need for a unified cross-border approach to disease control measures in the border areas. PMID:26516086

  5. Antimicrobial effect of chlorine dioxide on Actinobacillus actinomycetemcomitans in diabetes mellitus rats treated with insulin

    Tantin Ermawati


    Full Text Available Background: Periodontitis is a chronic inflammatory disease of periodontal tissues. Etiology of periodontal disease includes Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans which is the most predominant disease-causing bacteria found in the gingival sulcus. Periodontitis can be exacerbated by the systemic disease, such as diabetes mellitus considered as a metabolic disease characterized by hyperglycemia due to insulin deficiency. Treatment of periodontitis is then required in patients with type I diabetes to avoid radical reaction that can not only cause bleeding, but can also prevent infection, as a result, topical antimicrobial therapy and blood glucose control are required. Topical antimicrobial chlorine dioxide is a disinfectant that is effective in killing A. actinomycetemcomitans. Purpose: This study is aimed to determine the effects of topical antimicrobial chlorine dioxide gel or rinse on the number of A. actinomycetemcomitans in DM rats treated with insulin. Methods: 20 three month old male Wistar rats with weight of 170–200 grams were divided into four groups. First, periodontitis and DM were manipulated into all groups through aloksan injection with dose of 170 mg/kg. Those rats in group I were treated with insulin and chlorine dioxide gel, those in group II were treated with insulin and chlorine dioxide rinse, those in group III were treated with insulin only, and those in group IV were without treatment. In the third and seventh weeks, the number of A. actinomycetemcomitans was measured. The data was tested by using One-Way ANOVA test followed by LSD test. Results: The study showed that chlorine dioxide gel has a greater ability in reducing the number of A. actinomycetemcomitans than chlorine dioxide rinse although both are antimicrobials. Conclusion: It can be concluded that the use of chlorine dioxide gel can more effective to decrease the number of A. actinomycetemcomitans than chlorine dioxide rinse in DM rats

  6. Improved diagnostic medium for separation of Cryptococcus neoformans var. neoformans (serotypes A and D) and Cryptococcus neoformans var. gattii (serotypes B and C).

    Kwon-Chung, K J; Polacheck, I; Bennett, J E


    A simple new agar medium containing L-canavanine, glycine, and bromthymol blue was found to give a clearer and more accurate distinction between serotype A or D (Cryptococcus neoformans var. neoformans) and serotype B or C (C. neoformans var. gattii) than creatinine-dextrose-bromthymol blue or glycine-cycloheximide-phenol red media. Among 143 isolates of serotype A or D and 70 isolates of serotype B or C, the new medium correlated completely with the serotype, whereas nearly 11% of these isol...

  7. Production of latex agglutination reagents for pneumococcal serotyping

    Ortika Belinda D


    Full Text Available Abstract Background The current ‘gold standard’ for serotyping pneumococci is the Quellung test. This technique is laborious and requires a certain level of training to correctly perform. Commercial pneumococcal latex agglutination serotyping reagents are available, but these are expensive. In-house production of latex agglutination reagents can be a cost-effective alternative to using commercially available reagents. This paper describes a method for the production and quality control (QC of latex reagents, including problem solving recommendations, for pneumococcal serotyping. Results Here we describe a method for the production of latex agglutination reagents based on the passive adsorption of antibodies to latex particles. Sixty-five latex agglutination reagents were made using the PneuCarriage Project (PCP method, of which 35 passed QC. The other 30 reagents failed QC due to auto-agglutination (n=2, no reactivity with target serotypes (n=8 or cross-reactivity with non-target serotypes (n=20. Dilution of antisera resulted in a further 27 reagents passing QC. The remaining three reagents passed QC when prepared without centrifugation and wash steps. Protein estimates indicated that latex reagents that failed QC when prepared using the PCP method passed when made with antiserum containing ≤ 500 μg/ml of protein. Sixty-one nasopharyngeal isolates were serotyped with our in-house latex agglutination reagents, with the results showing complete concordance with the Quellung reaction. Conclusions The method described here to produce latex agglutination reagents allows simple and efficient serotyping of pneumococci and may be applicable to latex agglutination reagents for typing or identification of other microorganisms. We recommend diluting antisera or removing centrifugation and wash steps for any latex reagents that fail QC. Our latex reagents are cost-effective, technically undemanding to prepare and remain stable for long periods of

  8. Transplacental transmission of Bluetongue virus serotype 1 and serotype 8 in sheep: virological and pathological findings.

    Mirjam T W van der Sluijs

    Full Text Available The Bluetongue virus serotype 8 (BTV-8 strain, which emerged in Europe in 2006, had an unusually high ability to cause foetal infection in pregnant ruminants. Other serotypes of BTV had already been present in Europe for more than a decade, but transplacental transmission of these strains had never been demonstrated. To determine whether transplacental transmission is a unique feature of BTV-8 we compared the incidence and pathological consequences of transplacental transmission of BTV-8 to that of BTV-1. Nine pregnant ewes were infected with either BTV-8 or BTV-1. The BTV strains used for the infection were field strains isolated on embryonated chicken eggs and passaged twice on mammalian cells. Blood samples were taken to monitor the viraemia in the ewes. Four weeks after the infection, the foetuses were examined for pathological changes and for the presence of BTV. BTV-8 could be demonstrated in 12 foetuses (43% from 5 ewes (56%. %. BTV-1 was detected in 14 foetuses (82% from 6 ewes (67%. Pathological changes were mainly found in the central nervous system. In the BTV-8 group, lympho-histiocytic infiltrates, gliosis and slight vacuolation of the neuropil were found. BTV-1 infection induced a severe necrotizing encephalopathy and severe meningitis, with macroscopic hydranencephaly or porencephaly in 8 foetuses. In our experimental setting, using low passaged virus strains, BTV-1 was able to induce transplacental transmission to a higher incidence compared to BTV-8, causing more severe pathology.

  9. Genomics Reveals the Worldwide Distribution of Multidrug-Resistant Serotype 6E Pneumococci.

    van Tonder, Andries J; Bray, James E; Roalfe, Lucy; White, Rebecca; Zancolli, Marta; Quirk, Sigríður J; Haraldsson, Gunnsteinn; Jolley, Keith A; Maiden, Martin C J; Bentley, Stephen D; Haraldsson, Ásgeir; Erlendsdóttir, Helga; Kristinsson, Karl G; Goldblatt, David; Brueggemann, Angela B


    The pneumococcus is a leading pathogen infecting children and adults. Safe, effective vaccines exist, and they work by inducing antibodies to the polysaccharide capsule (unique for each serotype) that surrounds the cell; however, current vaccines are limited by the fact that only a few of the nearly 100 antigenically distinct serotypes are included in the formulations. Within the serotypes, serogroup 6 pneumococci are a frequent cause of serious disease and common colonizers of the nasopharynx in children. Serotype 6E was first reported in 2004 but was thought to be rare; however, we and others have detected serotype 6E among recent pneumococcal collections. Therefore, we analyzed a diverse data set of ∼1,000 serogroup 6 genomes, assessed the prevalence and distribution of serotype 6E, analyzed the genetic diversity among serogroup 6 pneumococci, and investigated whether pneumococcal conjugate vaccine-induced serotype 6A and 6B antibodies mediate the killing of serotype 6E pneumococci. We found that 43% of all genomes were of serotype 6E, and they were recovered worldwide from healthy children and patients of all ages with pneumococcal disease. Four genetic lineages, three of which were multidrug resistant, described ∼90% of the serotype 6E pneumococci. Serological assays demonstrated that vaccine-induced serotype 6B antibodies were able to elicit killing of serotype 6E pneumococci. We also revealed three major genetic clusters of serotype 6A capsular sequences, discovered a new hybrid 6C/6E serotype, and identified 44 examples of serotype switching. Therefore, while vaccines appear to offer protection against serotype 6E, genetic variants may reduce vaccine efficacy in the longer term because of the emergence of serotypes that can evade vaccine-induced immunity. PMID:25972423

  10. FDA Approves First Botulism Antitoxin for Use in Neutralizing All Seven Known Botulinum Nerve Toxin Serotypes

    ... Antitoxin for use in neutralizing all seven known botulinum nerve toxin serotypes Product to be stored in Strategic National ... antibody fragments that neutralize all of the seven botulinum nerve toxin serotypes known to cause botulism. Botulism is a ...

  11. Serotype-specific mortality from invasive Streptococcus pneumoniae disease revisited

    Martens, Pernille; Worm, Signe Westring; Lundgren, Bettina;


    with Streptococcus pneumoniae (pneumococci) causes significant morbidity and mortality. Case series and experimental data have shown that the capsular serotype is involved in the pathogenesis and a determinant of disease outcome. METHODS: Retrospective review of 464 cases of invasive disease among adults diagnosed......Serotype-specific mortality from invasive Streptococcus pneumoniae disease revisited.Martens P, Worm SW, Lundgren B, Konradsen HB, Benfield T. Department of Infectious Diseases 144, Hvidovre University Hospital, DK-2650 Hvidovre, Denmark. BACKGROUND: Invasive infection...... between 1990 and 2001. Multivariate Cox proportional hazard analysis. RESULTS: After adjustment for other markers of disease severity, we found that infection with serotype 3 was associated with an increased relative risk (RR) of death of 2.54 (95% confidence interval (CI): 1.22-5.27), whereas infection...

  12. Detection of Foot-and-mouth Disease Serotype O by ELISA Using a Monoclonal Antibody

    Chen, Hao-tai; Peng, Yun-hua; ZHANG, YONG-GUANG; Liu, Xiang-tao


    An ELISA assay with monoclonal antibody (MELISA) was used to type serotype O of foot-and-mouth disease virus (FMDV). All FMDV serotype O reference strains were positive by MELISA, while other viruses such as FMDV serotypes Asia 1, C, and A and classical swine fever virus, swine vesicular disease virus, and porcine reproductive and respiratory syndrome virus remained negative. Furthermore, FMDV serotype O positive samples were able to be detected by MELISA. This assay may be particularly suita...

  13. A mosaic adenovirus possessing serotype Ad5 and serotype Ad3 knobs exhibits expanded tropism

    The efficiency of cancer gene therapy with recombinant adenoviruses based on serotype 5 (Ad5) has been limited partly because of variable, and often low, expression by human primary cancer cells of the primary cellular-receptor which recognizes the knob domain of the fiber protein, the coxsackie and adenovirus receptor (CAR). As a means of circumventing CAR deficiency, Ad vectors have been retargeted by utilizing chimeric fibers possessing knob domains of alternate Ad serotypes. We have reported that ovarian cancer cells possess a primary receptor for Ad3 to which the Ad3 knob binds independently of the CAR-Ad5 knob interaction. Furthermore, an Ad5-based chimeric vector, designated Ad5/3, containing a chimeric fiber proteins possessing the Ad3 knob, demonstrates CAR-independent tropism by virtue of targeting the Ad3 receptor. Based on these findings, we hypothesized that a mosaic virus possessing both the Ad5 knob and the Ad3 knob on the same virion could utilize either primary receptor, resulting in expanded tropism. In this study, we generated a dual-knob mosaic virus by coinfection of 293 cells with Ad5-based and Ad5/3-based vectors. Characterization of the resultant virions confirmed the incorporation of both Ad5 and Ad3 knobs in the same particle. Furthermore, this mosaic virus was able to utilize either receptor, CAR and the Ad3 receptor, for virus attachment to cells. Enhanced Ad infectivity with the mosaic virus was shown in a panel of cell lines, with receptor profiles ranging from CAR-dominant to Ad3 receptor-dominant. Thus, this mosaic virus strategy may offer the potential to improve Ad-based gene therapy approaches by infectivity enhancement and tropism expansion

  14. Therapeutic effect of Chinese patent medicine "Wuhuanghu" on porcine infectious pleuropneumonia and its acute and subchronic toxicity as well as evaluation of safety pharmacology.

    Wang, Guangxi; Kang, Shuai; Yin, Zhongqiong; Jia, Ren-Yong; Lai, Xin; Zhou, Xun; Liang, Xiao-Xia; Li, Li-Xia; Zou, Yuan-Feng; Lv, Cheng; He, Chang-Liang; Ye, Gang; Yin, Li-Zi; Jing, Bo


    Chinese patent medicines play an important role in veterinary clinical use. The aim of this study is to research the anti-infection effect of Chinese patent medicine "Wuhuanghu" for the treatment of porcine infectious pleuropneumonia and to evaluate the safety of "Wuhuanghu" in order to provide a comprehensive understanding of its toxicity. The anti-infection results showed that the treatment with "Wuhuanghu" could significantly inhibit pneumonia and decrement of the pneumonia in high, medium and low doses of "Wuhuanghu" groups were 70.97%, 61.29% and 58.06% respectively. The acute toxicity test showed that rats in the highest group (5000mg/kg) had no death and no abnormal response, suggesting the LD50 of "Wuhuanghu" was more than 5000mg/kg. The subchronic toxicity study showed that hematology indexes in all groups had no obvious differences; blood biochemical index, only albumin and total cholesterol in middle and low doses of "Wuhuanghu" groups were significantly decreased when compared with control group. The clinical pathology showed that the target organ of "Wuhuanghu" was liver. The safety pharmacology study indicated that "Wuhuanghu" had no side effects on rats. In conclusion, "Wuhuanghu" has therapeutic and protective effects to porcine infectious pleuropneumonia in a dose-dependent manner and "Wuhuanghu" is a safe veterinary medicine. PMID:26245812

  15. Allergic reactions in salmonellosis depends on the Serotype of pathogens

    Mkrtchyan, M.S.; Zakaryan, М. K.; Mnatsakanyan, А. А.;


    .Enteritidis). Previously, we reported that the induction of the cytokine network and an antimicrobial protein is serotype-specific and also depends on the disease stage. Differential genomic context of the serotypes may explain the differential induction of inflammatory responses. Recent studies have indicated that...... bacterial infections in early life may help to inhibit excessive allergic Th2 reactions by angling the immune system towards Th1 responses. However, it is known that infections can also cause the exacerbation of allergic reactions. Skewing of response away from Treg cells may lead to the onset and...

  16. Novel Pneumococcal Serotypes 6C and 6D: Anomaly or Harbinger

    McEllistrem, M. Catherine; Nahm, Moon H.


    The discovery of serotypes 6C and 6D, the former of which expanded in the 7-valent pneumococcal protein conjugate vaccine era, illustrates a previously unrecognized limitation of conjugate vaccines: the expansion of unrecognized serotypes could erode the efficacy of a serotype-specific vaccine.

  17. Genomic Evolution Of The Mdr Serotype O12 Pseudomonas Aeruginosa Clone

    Thrane, Sandra Wingaard; Taylor, Véronique L.; Freschi, Luca;


    that serotype switching in combination with an antibiotic resistance determinant contributed to the dissemination of the O12 serotype in the clinic. This selective advantage coincides with the introduction of fluoroquinolones in the clinic. With the PAst program isolates can be serotyped using WGS data...

  18. Multidrug resistance among different serotypes of clinical Salmonella isolates in Taiwan

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


    (41%) and was highly prevalent in Salmonella enterica serotype Typhimurium (72.7%, 176/242) the most common serotype. Additional resistance to trimethoprim was present in 155 (19.4% overall) of the ACSSuT R-type isolates from several serotypes. Reduced susceptibility to fluoroquinolone (FQ...

  19. MicroRNA expression in lung tissue and blood isolated from pigs suffering from bacterial pneumonia

    Skovgaard, Kerstin; Wendt, Karin Tarp; Heegaard, Peter M. H.

    immune responses. Still, little is known about the impact of miRNAs in the development and pathogenesis of lung infections. Expression of miRNA, known to be induced by bacterial (i.e., LPS) ligands and thus supposed to play a role in the regulation of antimicrobial defence, were studied in lung tissue...... all miRNAs for human, mouse and rat. The miRCURY™ LNA array microarray slides were scanned, and image analysis was carried out using the ImaGene 8.0 software (BioDiscovery, Inc., USA). A two-tailed T-test calculated between infected and control identified 10 of 1263 miRNA to be differentially...... from pigs experimentally infected with Actinobacillus pleuropneumoniae serotype 2 and 6. Circulating miRNAs were studied in blood from pigs infected with A. pleuropneumoniae serotype 2 using real time-qPCR (RT-qPCR). Expression profiles of miRNA in blood of seven animals before and after infection...

  20. Bagasse hydrolyzates from Agave tequilana as substrates for succinic acid production by Actinobacillus succinogenes in batch and repeated batch reactor.

    Corona-González, Rosa Isela; Varela-Almanza, Karla María; Arriola-Guevara, Enrique; Martínez-Gómez, Álvaro de Jesús; Pelayo-Ortiz, Carlos; Toriz, Guillermo


    The aim of this work was to obtain fermentable sugars by enzymatic or acid hydrolyses of Agave tequilana Weber bagasse in order to produce succinic acid with Actinobacillus succinogenes. Hydrolyses were carried out with mineral acids (sulfuric and hydrochloric acids) or a commercial cellulolytic enzyme, and were optimized statistically by a response surface methodology, having as factors the concentration of acid/enzyme and time of hydrolysis. The concentration of sugars obtained at optimal conditions for each hydrolysis were 21.7, 22.4y 19.8g/L for H2SO4, HCl and the enzymatic preparation respectively. Concerning succinic acid production, the enzymatic hydrolyzates resulted in the highest yield (0.446g/g) and productivity (0.57g/Lh) using A. succinogenes in a batch reactor system. Repeated batch fermentation with immobilized A. succinogenes in agar and with the enzymatic hydrolyzates resulted in a maximum concentration of succinic acid of 33.6g/L from 87.2g/L monosaccharides after 5 cycles in 40h, obtaining a productivity of 1.32g/Lh. PMID:26802183

  1. Performance analyses of a neutralizing agent combination strategy for the production of succinic acid by Actinobacillus succinogenes ATCC 55618.

    Wang, Cheng-Cheng; Zhu, Li-Wen; Li, Hong-Mei; Tang, Ya-Jie


    A neutralizing agent combination strategy was developed to enhance the succinic acid production by Actinobacillus succinogenes ATCC 55618. First, a maximal succinic acid production of 48.2 g/L was obtained at a culture pH of 7.5. Second, NaOH and KOH were screened to identify the optimal neutralizing agent for pH control. However, the production of succinic acid did not increase, and severe cell flocculation was observed due to a high concentration of metal ions when only one neutralizing agent was used to control pH. Finally, a neutralizing agent combination strategy was developed with a supply of neutralizing agents with OH(-) and carbonate. The cell flocculation was eliminated, and a maximum succinic acid production of 59.2 g/L was obtained with 5 M NaOH and 40 g/L of MgCO(3); this production was 27.9% higher than that obtained with NaOH alone. The results obtained in this study may be useful for the large-scale industrial production of succinic acid. PMID:22002101

  2. Actinobacillus succinogenes ATCC 55618 fermentation medium optimization for the production of succinic acid by response surface methodology.

    Zhu, Li-Wen; Wang, Cheng-Cheng; Liu, Rui-Sang; Li, Hong-Mei; Wan, Duan-Ji; Tang, Ya-Jie


    As a potential intermediary feedstock, succinic acid takes an important place in bulk chemical productions. For the first time, a method combining Plackett-Burman design (PBD), steepest ascent method (SA), and Box-Behnken design (BBD) was developed to optimize Actinobacillus succinogenes ATCC 55618 fermentation medium. First, glucose, yeast extract, and MgCO(3) were identified to be key medium components by PBD. Second, preliminary optimization was run by SA method to access the optimal region of the key medium components. Finally, the responses, that is, the production of succinic acid, were optimized simultaneously by using BBD, and the optimal concentration was located to be 84.6 g L(-1) of glucose, 14.5 g L(-1) of yeast extract, and 64.7 g L(-1) of MgCO(3). Verification experiment indicated that the maximal succinic acid production of 52.7 ± 0.8 g L(-1) was obtained under the identified optimal conditions. The result agreed with the predicted value well. Compared with that of the basic medium, the production of succinic acid and yield of succinic acid against glucose were enhanced by 67.3% and 111.1%, respectively. The results obtained in this study may be useful for the industrial commercial production of succinic acid. PMID:23093852

  3. Use of corn steep liquor as an economical nitrogen source for biosuccinic acid production by Actinobacillus succinogenes

    Tan, J. P.; Jahim, J. M.; Wu, T. Y.; Harun, S.; Mumtaz, T.


    Expensive raw materials are the driving force that leads to the shifting of the petroleum-based succinic acid production into bio-based succinic acid production by microorganisms. Cost of fermentation medium is among the main factors contributing to the total production cost of bio-succinic acid. After carbon source, nitrogen source is the second largest component of the fermentation medium, the cost of which has been overlooked for the past years. The current study aimed at replacing yeast extract- a costly nitrogen source with corn steep liquor for economical production of bio-succinic acid by Actinobacillus succinogenes 130Z. In this study, a final succinic acid concentration of 20.6 g/L was obtained from the use of corn steep liquor as the nitrogen source, which was comparable with the use of yeast extract as the nitrogen source that had a final succinate concentration of 21.4 g/l. In terms of economical wise, corn steep liquor was priced at 200 /ton, which was one fifth of the cost of yeast extract at 1000 /ton. Therefore, corn steep liquor can be considered as a potential nitrogen source in biochemical industries instead of the costly yeast extract.

  4. Transcriptional profiling at different sites in lungs of pigs during acute bacterial respiratory infection

    Mortensen, Shila; Skovgaard, Kerstin; Hedegaard, Jakob; Bendixen, Christian; Heegaard, Peter M. H.


    The local transcriptional response was studied in different locations of lungs from pigs experimentally infected with the respiratory pathogen Actinobacillus pleuropneumoniae serotype 5B, using porcine cDNA microarrays. This infection gives rise to well-demarcated infection loci in the lung...... apoptosis and the complement system. Interferon-g was downregulated in both necrotic and bordering areas. Evidence of neutrophil recruitment was seen by the up-regulation of chemotactic factors for neutrophils. In conclusion, we found subsets of genes expressed at different levels in the three selected...... of induced genes as, in unaffected areas a large part of differently expressed genes were involved in systemic reactions to infections, while differently expressed genes in necrotic areas were mainly concerned with homeostasis regulation....

  5. A comparison of salmonella serotypes found in the faeces of gulls feeding at a sewage works with serotypes present in the sewage.

    Fenlon, D.R.


    The numbers of salmonella serotypes in raw sewage, sewage sludge and final effluent at a sewage treatment works were determined. Resting gulls which had previously been feeding on the sewage were disturbed and individual faecal samples tested for the presence of salmonellae. The serotypes were compared with those in the sewage. Six serotypes were isolated from the sewage, Salmonella stanley being found in all types of samples. Eleven of the twenty gull faeces samples were positive for salmone...

  6. PCR-Based Serotyping of Streptococcus pneumoniae from Culture-Negative Specimens: Novel Primers for Detection of Serotypes within Serogroup 18.

    Tanmoy, Arif M; Saha, Senjuti; Darmstadt, Gary L; Whitney, Cynthia G; Saha, Samir K


    Six multiplex-compatible PCR primers were designed to distinguish Streptococcus pneumoniae serotypes within serogroup 18 from culturable/nonculturable pneumococcal specimens, with no cross-reactivity with other serotypes and respiratory organisms. These primers will aid in the generation of better data on vaccine/nonvaccine serotypes in invasive and carriage pneumococcal surveillance and contribute to future vaccine formulation and impact studies. PMID:27252464

  7. Ceftriaxone-resistant Salmonella enterica serotype Newport, France

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


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

  8. Ceftriaxone-Resistant Salmonella enterica Serotype Newport, France

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


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

  9. Urban Epidemic of Dengue Virus Serotype 3 Infection, Senegal, 2009

    Faye, Ousmane; Ba, Yamar; Faye, Oumar; Talla, Cheikh; Diallo, Diawo; Chen, Rubing; Mondo, Mireille; Ba, Rouguiétou; Macondo, Edgard; Siby, Tidiane; Weaver, Scott C.; Diallo, Mawlouth; Sall, Amadou Alpha


    An urban epidemic of dengue in Senegal during 2009 affected 196 persons and included 5 cases of dengue hemorrhagic fever and 1 fatal case of dengue shock syndrome. Dengue virus serotype 3 was identified from all patients, and Aedes aegypti mosquitoes were identified as the primary vector of the virus.

  10. Reevaluating the Serotype II Capsular Locus of Streptococcus agalactiae▿

    Martins, E. R.; Melo-Cristino, J.; Ramirez, M.


    We report a novel sequence of the serotype II capsular locus of group B streptococcus that resolves inconsistencies among the results of various groups and the sequence in GenBank. This locus was found in diverse lineages and presents genes consistent with the complete synthesis of the type II polysaccharide.

  11. Selection of a screening panel of rhinoviral serotypes.

    Tomkinson, Nick; Wenlock, Mark; McCrae, Christopher


    A novel methodology for the selection of a representative primary and secondary screening panel of rhinoviral serotypes for the purposes of identifying potential antiviral agents is presented. This methodology focuses on the active-sites of the rhinoviral proteins but does not invoke historical SAR data, thereby avoiding compound bias. PMID:23122823

  12. Foot-and-Mouth Disease Virus Serotype A in Egypt

    Knowles, Nick J.; Wadsworth, Jemma; Reid, Scott M; Swabey, Katherine G.; El-Kholy, Alaa A.; El-Rahman, Adel Omar Abd; Soliman, Hatem M.; Ebert, Katja; Ferris, Nigel P.; Hutchings, Geoffrey H.; Statham, Robert J.; King, Donald P.; Paton, David J.


    We describe the characterization of a foot-and-mouth disease (FMD) serotype A virus responsible for recent outbreaks of disease in Egypt. Phylogenetic analysis of VP1 nucleotide sequences demonstrated a close relationship to recent FMD virus isolates from East Africa, rather than to viruses currently circulating in the Middle East.

  13. Clonal distribution of pneumococcal serotype 19F isolates from Ghana

    Sparding, Nadja; Dayie, Nicholas Tete Kwaku Dzifa; Mills, Richael O.;


    Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. Pneumococcal strains are classified according to their capsular polysaccharide and more than 90 different serotypes are currently known. In this project, three distinct groups of pneumococcal carriage isolates from Gh...

  14. Detection of foot-and-mouth disease serotype O by ELISA using a monoclonal antibody.

    Chen, Hao-Tai; Peng, Yun-Hua; Zhang, Yong-Guang; Liu, Xiang-Tao


    An ELISA assay with monoclonal antibody (MELISA) was used to type serotype O of foot-and-mouth disease virus (FMDV). All FMDV serotype O reference strains were positive by MELISA, while other viruses such as FMDV serotypes Asia 1, C, and A and classical swine fever virus, swine vesicular disease virus, and porcine reproductive and respiratory syndrome virus remained negative. Furthermore, FMDV serotype O positive samples were able to be detected by MELISA. This assay may be particularly suitable for diagnosis of FMDV serotype O infection in field stations. PMID:23600506

  15. Group B streptococcus serotype prevalence in reproductive-age women at a tertiary care military medical center relative to global serotype distribution

    Williams Julie


    Full Text Available Abstract Background Group B Streptococcus (GBS serotype (Ia, Ib, II-IX correlates with pathogen virulence and clinical prognosis. Epidemiological studies of seroprevalence are an important metric for determining the proportion of serotypes in a given population. The purpose of this study was to evaluate the prevalence of individual GBS serotypes at Madigan Healthcare System (Madigan, the largest military tertiary healthcare facility in the Pacific Northwestern United States, and to compare seroprevalences with international locations. Methods To determine serotype distribution at Madigan, we obtained GBS isolates from standard-of-care anogenital swabs from 207 women of indeterminate gravidity between ages 18-40 during a five month interval. Serotype was determined using a recently described molecular method of polymerase chain reaction by capsular polysaccharide synthesis (cps genes associated with pathogen virulence. Results Serotypes Ia, III, and V were the most prevalent (28%, 27%, and 17%, respectively. A systematic review of global GBS seroprevalence, meta-analysis, and statistical comparison revealed strikingly similar serodistibution at Madigan relative to civilian-sector populations in Canada and the United States. Serotype Ia was the only serotype consistently higher in North American populations relative to other geographic regions (p Conclusion This study establishes PCR-based serotyping as a viable strategy for GBS epidemiological surveillance. Our results suggest that GBS seroprevalence remains stable in North America over the past two decades.

  16. Implementation of Salmonella serotype determination using pulsed-field gel electrophoresis in a state public health laboratory.

    Bopp, Dianna J; Baker, Deborah J; Thompson, Lisa; Saylors, Amy; Root, Timothy P; Armstrong, Leeanna; Mitchell, Kara; Dumas, Nellie B; Musser, Kimberlee Arruda


    We examined the use of pulsed-field gel electrophoresis (PFGE) to predict serotype for Salmonella isolates. Between 2012 and 2014 we assessed 4481 isolates, resulting in >90% assigned serotypes. PFGE is efficient for determining serotype in the majority of cases and results in expedited serotype determination, as well as cost savings. PMID:27220605

  17. Serotype Distribution of Salmonella Isolates from Turkey Ground Meat and Meat Parts

    Irfan Erol


    Full Text Available The aim of the study was to find out the serotype distribution of 169 Salmonella colonies recovered from 112 Salmonella positive ground turkey (115 colonies and 52 turkey meat parts (54 colonies. Out of 15 Salmonella serotypes: S. Corvallis, S. Kentucky, S. Bredeney, S. Virchow, S. Saintpaul and S. Agona were identified as the predominant serovars at the rates of 27%, 13%, 12%, 12%, 11%, and 10%, respectively. Other serotypes were below 6% of the total isolates. All S. Kentucky and S. Virchow and most of the S. Corvallis (39/46 and S. Heidelberg (9/9 serotypes were recovered from ground turkey. The results indicate that turkey ground meat and meat parts were contaminated with quite distinct Salmonella serotypes. This is the first study reporting Salmonella serotype distribution in turkey meat and S. Corvallis as predominant serotype in poultry meat in Turkey.

  18. Dicty_cDB: Contig-U05198-1 [Dicty_cDB

    Full Text Available e) Actinobacillus pleuropneumoniae ... 49 6e-08 CP001100_1460( CP001100 |pid:none) Chloroherpeton thalassi...bacterium freudenreichii ... 44 2e-08 CP000687_808( CP000687 |pid:none) Actinobacillus pleuropneumoni...000569_834( CP000569 |pid:none) Actinobacillus pleuropneumoniae ... 47 2e-07 CP000951_1090( CP000951 |pid:non...01213_1384( CP001213 |pid:none) Bifidobacterium animalis subsp.... 42 6e-07 CP000142_2488( CP000142 |pid:none) Pelobacter carbino...e) Chlorobaculum parvum NCIB 8327, ... 57 9e-07 AY625502_1( AY625502 |pid:none) Glossina morsitans morsitans clon

  19. LEUKOTOXIC ACTIVITY OF ACTINOBACILLUS ACTINOMYCETEMCOMITANS ISOLATED FROM HUMAN AND NON-HUMAN PRIMATES Atividade leucotóxica de amostras de Actinobacillus actinomycetemcomitans de primatas humanos não-humanos

    Francisca Lúcia de Lima


    Full Text Available Actinobacillus actinomycetemcomitans is a clinically relevant periodontopathogenic Gram-negative coccobacillus that produces a leukotoxin of the RTX cytolysin family. In this study, we evaluated the leukotoxic activity of A. actinomycetemcomitans strains isolated from human and marmosets by Trypan blue exclusion and by the chemiluminescence assays. Among eight A. actinomycetemcomitans human strains studied, two (P2.17 and P8.12 were classified as high leukotoxin producers and among eight marmoset strains, one (M22.11 showed high leukotoxin production, as determined by Trypan blue exclusion assay. The reference strains ATCC 29523 and FDC Y4 respectively behaved like moderate and low producers. The chemiluminescence assay was used to evaluate the leukotoxic activity of M22.11 and P2.17 strains submitted to different growth conditions. Leukotoxic activity was detected on cells at the logarithmic phase and was similar under anaerobic and microaerophilic growth conditions. It was greatly reduced when cells were grown at glucose concentrations lower or higher than 0.75% (0.25% and 1.5% in thioglycolate medium. Leukotoxin production mainly by the M22.11 strain was low in BHI broth, whereas production in TSB medium showed a similar level as in thioglycolate broth medium. Sodium bicarbonate at 10 mM did not affect leukotoxin production.Actinobacillus actinomycetemcomitans é um cocobacilo Gram negativo, periodontopatógeno clinicamente importante, que produz uma leucotoxina pertencente à família das citolisinas RTX. Neste estudo, avaliou-se a atividade leucotóxica de amostras de A. actinomycetemcomitans isoladas de seres humanos e de calitriquídeos pelos métodos de exclusão de azul de Tripan e quimioluminescência. Duas (P2.17 e P8.12 entre oito amostras de A. actinomycetemcomitans isoladas de seres humanos, e uma (M22.11 entre 8 amostras isoladas de sagüis se apresentaram como altamente produtoras de leucotoxina, como determinado pelo teste de

  20. Impact of pneumococcal vaccination in children on serotype distribution in adult community-acquired pneumonia using the serotype-specific multiplex urinary antigen detection assay.

    Pletz, Mathias W; Ewig, Santiago; Rohde, Gernot; Schuette, Hartwig; Rupp, Jan; Welte, Tobias; Suttorp, Norbert; Forstner, Christina


    The aim of the study was to compare the distribution of the vaccine-serotypes covered by pneumococcal conjugate vaccines (PCV7 and PCV13) in adult patients with pneumococcal community-acquired pneumonia in Germany between the periods 2002-2006 and 2007-2011 using a novel serotype-specific multiplex urinary antigen detection assay (SSUA). Vaccination of children started with PCV7 in 2007, which was replaced by PCV13 in 2010. Following confirmation of the accuracy of SSUA in long-term stored urine samples from 112 patients with confirmed pneumonia and known pneumococcal serotype, urine samples of 391 CAPNETZ patients with documented pneumococcal pneumonia (i.e. positive BinaxNOW(®) Streptococcus pneumoniae urine antigen test) but unknown serotype were tested for the 13 vaccine-serotypes using SSUA. The proportion of PCV7-serotypes significantly decreased in adult patients with pneumonia from 30.6% (2002-6) to 13.3% (2007-11, ppneumococcal serotypes included by PCV13 remained stable during study period with a coverage of 61.5% (2002-06) and 59.7% (2007-11) in non-bacteremic pneumonia and 79% (for both periods) in bacteremic pneumonia, mainly due to an increase in pneumococcal serotypes 1, 3 and 7F during the second period. Thus, implementation of PCV7 in children in Germany in 2007 was associated with a significant decrease in vaccine-serotypes covered by PCV7 in adult patients with non-bacteremic pneumococcal pneumonia and with an elimination of PCV7 vaccine-serotypes in bacteremic pneumococcal pneumonia. PCV13 coverage remained high up to 2011, mainly due to an increase in serotypes 1, 3 and 7F. German Clinical Trials Register: DRKS00005274. PMID:27016653

  1. Association between coinfection of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Treponema denticola and periodontal tissue destruction in chronic periodontitis

    CHEN Li-li; WU Yan-min; YAN Jie; SUN Wei-lian; SUN Yu-zheng; David Ojcius


    Background The association between the infection of Porphyromonas gingivalis, Actinobacillus actinomy-cetemcomitans and Treponema denticola in chronic periodontitis (CP) and the severity of periodontal disease remains to be elucidated. The aim of this study was to investigate the subgingival infection frequencies of three periodontopathic bacteria in Chinese CP patients and to evaluate the correlations between infection by these bacteria and periodontal destruction.Methods A multiple PCR assay using primers derived from 16SrDNA genes of P. gingivalis, A. actinomy-cetemitans and T. denticola was established to measure simultaneously the presence of the three microbes in 162 subgingival samples from 81 Chinese CP patients. Results The positive rates of P. gingivalis, A. actinomycetemitans and T. denticola in the subgingival samples were 84.6%, 83.3% and 88.3%, respectively. Of the subgingival samples, 68% revealed the coinfection of all the three microbes. The infection rates with P. gingivalis, A. actinomycetemitans or T. denticola alone was 5.9% (1/17), 17.6% (3/17) and 76.5% (13/17), respectively. A close association was present between the A. actinomycetemitans infection and gingival index (GI) (P0.05). P. gingivalis and A. actinomycetemitans were more frequently detectable in middle and deep pockets than in shallow ones (P<0.01), while T. denticola was found remarkably often in deep pockets (P<0.05). The coinfection rate of the three microbes was significantly higher in sites with severe periodontitis than in those with mild periodontitis (P<0.01). Conclusions The multiple PCR established in this study can be used as a sensitive and specific method to simultaneously detect all three microbes in subgingival samples. A. actinomycetemitans infection may be associated with CP and play an important role in the periodontal tissue destruction. The coinfection of P. gingivalis, A. actinomycetemitans and T. denticola can cause more serious periodontal destruction than

  2. Coinfection by Ureaplasma spp., Photobacterium damselae and an Actinomyces-like microorganism in a bottlenose dolphin (Tursiops truncatus) with pleuropneumonia stranded along the Adriatic coast of Italy.

    Di Francesco, Gabriella; Cammà, Cesare; Curini, Valentina; Mazzariol, Sandro; Proietto, Umberto; Di Francesco, Cristina Esmeralda; Ferri, Nicola; Di Provvido, Andrea; Di Guardo, Giovanni


    A case of pleuropneumonia is reported in an adult male bottlenose dolphin (Tursiops truncatus) found stranded in 2014 along the Central Adriatic coast of Italy. A severe pyogranulomatous pneumonia and thoracic lymphadenopathy were present at necropsy. Numerous Splendore-Hoeppli bodies were found microscopically scattered throughout the lung. Histochemical evidence of Actinomyces-like organisms was obtained from the pulmonary parenchyma, with a strain of Photobacterium damselae subsp. piscicida and Ureaplasma spp. being also isolated from the same tissue. For the latter, a genome fragment of approximately 1400bp from the 16s rDNA was amplified and sequenced. BLAST analysis revealed 100% identity with an uncultured Ureaplasma spp. (JQ193826.1). PMID:27033917

  3. Draft Genome Sequences of Listeria monocytogenes Serotype 4b Strains 944 and 2993 and Serotype 1/2c Strains 198 and 2932

    Casey, Aidan; Fox, Edward M.; Leong, Dara; Gahan, Cormac G. M.; Jordan, Kieran


    Listeria monocytogenes is a foodborne pathogen and the causative agent of listeriosis among humans and animals. The draft genome sequences of L. monocytogenes serotype 4b strains 944 and 2993 and serotype 1/2c strains 198 and 2932 are reported here. PMID:27257200

  4. Serotype-specific differences in short- and longer-term mortality following invasive pneumococcal disease.

    Hughes, G J; Wright, L B; Chapman, K E; Wilson, D; Gorton, R


    Invasive pneumococcal disease (IPD), caused by infection with Streptococcus pneumoniae, has a substantial global burden. There are over 90 known serotypes of S. pneumoniae with a considerable body of evidence supporting serotype-specific mortality rates immediately following IPD. This is the first study to consider the association between serotype and longer-term mortality following IPD. Using enhanced surveillance data from the North East of England we assessed both the short-term (30-day) and longer-term (⩽7 years) independent adjusted associations between individual serotypes and mortality following IPD diagnosis using logistic regression and extended Cox proportional hazards models. Of the 1316 cases included in the analysis, 243 [18·5%, 95% confidence interval (CI) 16·4-20·7] died within 30 days of diagnosis. Four serotypes (3, 6A, 9N, 19 F) were significantly associated with overall increased 30-day mortality. Effects were observable only for older adults (⩾60 years). After extension of the window to 12 months and 36 months, one serotype was associated with significantly increased mortality at 12 months (19 F), but no individual serotypes were associated with increased mortality at 36 months. Two serotypes had statistically significant hazard ratios (HR) for longer-term mortality: serotype 1 for reduced mortality (HR 0·51, 95% CI 0·30-0·86) and serotype 9N for increased mortality (HR 2·30, 95% CI 1·29-4·37). The association with serotype 9N was no longer observed after limiting survival analysis to an observation period starting 30 days after diagnosis. This study supports the evidence for associations between serotype and short-term (30-day) mortality following IPD and provides the first evidence for the existence of statistically significant associations between individual serotypes and longer-term variation in mortality following IPD. PMID:27193457

  5. Study on Serotypes and Auxotypes of Neisseria Gonorrhoeae in Guangzhou

    ZHENG Heping(郑和平); PAN Huiqing(潘慧清); HUANG Jinmei(黄进梅); ZENG Weiying(曾维英); WU Xingzhong(吴兴中); LIU Zhongqiu(刘仲秋)


    Objective: To investigate the serotypes and auxotypesdistribution of Neisseria gonorrhoeae in Guangzhou.Method: 131 strains of Neisseria gonorrhoeae wereserotyped by co-agglutination test and 108 strains wereauxotyped by La Scolea′s method.Results: Out of 131 strains of Neisseria gonorrhoeae ,87.8% (115/131) were WⅡ/WⅢ, while 9.9% (13/131) wereWI. The most important auxotypes were Proto, Pro and ILe,42.6% (46/108), 21.3% (23/108) and 12.0%, respectively. WⅡ/WⅢ was distributed among the all auxotypes aboveand WI found only in both Proto and Pro.Conclusion: The study illustrated the prevailing serotype,WⅡ/WⅢ, and higher prevalence of Ile- in Guangzhou.

  6. Inflammatory Responses to Salmonella Infections Are Serotype-Specific.

    Ktsoyan, Zhanna; Ghazaryan, Karine; Manukyan, Gayane; Martirosyan, Anush; Mnatsakanyan, Armine; Arakelova, Karine; Gevorgyan, Zaruhi; Sedrakyan, Anahit; Asoyan, Ara; Boyajyan, Anna; Aminov, Rustam


    The main purpose of this study was to investigate the profile of inflammatory response in patients with acute salmonellosis caused by two serotypes of Salmonella enterica, S. Enteritidis and S. Typhimurium, as well as in convalescent patients with previous acute disease caused by S. Enteritidis. Patients with acute disease showed significantly elevated levels of IL-1β, IL-17, IL-10, and calprotectin compared to healthy control subjects. In convalescent patients, these markers were also significantly elevated, with the exception of IL-1β. Multivariate statistical analyses with the use of these variables produced models with a good predictive accuracy resulting in excellent separation of the diseased and healthy cohorts studied. Overall, the results suggest that the profile of inflammatory response in this disease is determined, to a significant degree, by the serotype of Salmonella, and the profile of certain cytokines and calprotectin remains abnormal for a number of months following the acute disease stage. PMID:26904722

  7. All Serotypes of Dengue Viruses Circulating in Kuala Lumpur, Malaysia

    M.H. Chew


    Full Text Available Dengue is a severe disease caused by dengue virus (DENV, transmitted to human being by infected Aedes mosquitoes. It is a major public health concern in Southeast Asia due to its fatality in the form of hemorrhagic fever (DHF and dengue shock syndrome (DSS. The objective of the study was to isolate and identify dengue virus serotypes prevalent in endemic areas of Kuala Lumpur and Selangor in Malaysia by virus culture, indirect immunoflurecent assay and molecular techniques. A total number of 232 sera samples were obtained from patients with clinical manifestations of dengue fever reported to University Kebangsaan Malaysia Medical Centre (UKMMC. The sera samples collected, were analyzed for IgM/IgG detection for the assessment of primary and secondary dengue fever, propagation in cell-line C36/36, Indirect Immunoflurecent Assay (IFA and RT-PCR. The study confirmed 46 dengue cases where 15 (32.61% were dual infections with DENV-1 and DENV- 4, 12 (26.09% dual infections with DENV-3 and DENV-4, and 11 (23.91% were dual infection with DENV-2 and DENV-4. Only 1 (2.17% was dengue infection with DENV-3 and 7 (15.22% were with DENV-4. Dengue serotype 4 was the most common serotype identified in the present study .The highest number of dengue cases detected in Cheras, Kuala Lumpur where all 4 types of dengue virus were prevalent. All serotypes of dengue viruses circulation only in Kuala Lumpur and Selangor Malaysia, needs further strengthening of the dengue preventive measure in the city areas and in the country.

  8. All Serotypes of Dengue Viruses Circulating in Kuala Lumpur, Malaysia

    M.H. Chew; Rahman, M. M.; J. Jelip; M. R. Hassan; Isahak, I.


    Dengue is a severe disease caused by dengue virus (DENV), transmitted to human being by infected Aedes mosquitoes. It is a major public health concern in Southeast Asia due to its fatality in the form of hemorrhagic fever (DHF) and dengue shock syndrome (DSS). The objective of the study was to isolate and identify dengue virus serotypes prevalent in endemic areas of Kuala Lumpur and Selangor in Malaysia by virus culture, indirect immunoflurecent assay and molecular techniques. A total number ...

  9. Serotype analysis of rotaviruses from different locations in Malaysia.

    Rasool, N B; Green, K Y; Kapikian, A Z


    The distribution of rotavirus G (VP7) serotypes circulating in four locations in Malaysia, representing three geographical areas, was evaluated in 341 RNA-positive stool specimens obtained discontinuously between 1977 and 1988 from infants and young children under the age of five years who were hospitalized with acute gastroenteritis. A total of 306 specimens (256 stool suspensions and 50 that were adapted to growth in tissue culture) that were rotavirus positive by the confirmatory enzyme-li...

  10. Inflammatory Responses to Salmonella Infections Are Serotype-Specific

    Anna Boyajyan; Ara Asoyan; Anahit Sedrakyan; Zaruhi Gevorgyan; Karine Arakelova; Armine Mnatsakanyan; Anush Martirosyan; Gayane Manukyan; Karine Ghazaryan; Zhanna Ktsoyan; Rustam Aminov


    The main purpose of this study was to investigate the profile of inflammatory response in patients with acute salmonellosis caused by two serotypes of Salmonella enterica, S. Enteritidis and S. Typhimurium, as well as in convalescent patients with previous acute disease caused by S. Enteritidis. Patients with acute disease showed significantly elevated levels of IL-1β, IL-17, IL-10, and calprotectin compared to healthy control subjects. In convalescent patients, these markers were also signif...

  11. Increasing quinolone resistance in Salmonella enterica serotype enteritidis

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


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

  12. Serotype Chimeric Human Adenoviruses for Cancer GeneTherapy

    Akseli Hemminki


    Full Text Available Cancer gene therapy consists of numerous approaches where the common denominator is utilization of vectors for achieving therapeutic effect. A particularly potent embodiment of the approach is virotherapy, in which the replication potential of an oncolytic virus is directed towards tumor cells to cause lysis, while normal cells are spared. Importantly, the therapeutic effect of the initial viral load is amplified through viral replication cycles and production of progeny virions. All cancer gene therapy approaches rely on a sufficient level of delivery of the anticancer agent into target cells. Thus,enhancement of delivery to target cells, and reduction of delivery to non-target cells, in an approach called transductional targeting, is attractive. Both genetic and non-genetic retargeting strategies have been utilized. However, in the context of oncolytic viruses, it is beneficial to have the specific modification included in progeny virions and hence genetic modification may be preferable. Serotype chimerism utilizes serotype specific differences in receptor usage, liver tropism and seroprevalence in order to gain enhanced infection of target tissue. This review will focus on serotype chimeric adenoviruses for cancer gene therapy applications.

  13. Experimental Contagious Caprine Pleuropneumonia: A Long Term Study on the Course of Infection and Pathology in a Flock of Goats Infected with Mycoplasma capricolum subsp. capripneumoniae

    Bölske G; Thiaucourt F; Wesonga HO; Wanjohi C; Lindberg R


    Contagious caprine pleuropneumonia (CCPP) is a major threat to goat farming in parts of Africa and Asia. It classically causes acute high morbidity and mortality early in infection, but little is known of its long term epizootiology and course. In this study, 10 goats were inoculated with Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and then mixed with 15 goats for contact transmission. The disease course was monitored in each goat for 56–105 days, whereafter the goats w...

  14. A DNA Microarray-Based Assay to Detect Dual Infection with Two Dengue Virus Serotypes

    Alvaro Díaz-Badillo


    Full Text Available Here; we have described and tested a microarray based-method for the screening of dengue virus (DENV serotypes. This DNA microarray assay is specific and sensitive and can detect dual infections with two dengue virus serotypes and single-serotype infections. Other methodologies may underestimate samples containing more than one serotype. This technology can be used to discriminate between the four DENV serotypes. Single-stranded DNA targets were covalently attached to glass slides and hybridised with specific labelled probes. DENV isolates and dengue samples were used to evaluate microarray performance. Our results demonstrate that the probes hybridized specifically to DENV serotypes; with no detection of unspecific signals. This finding provides evidence that specific probes can effectively identify single and double infections in DENV samples.

  15. Emergence of group B Streptococcus serotype IV in women of child-bearing age in Ireland.

    Kiely, R A


    This study determined the carriage rate and serotype distribution of group B Streptococcus (GBS) in women of child-bearing age in the southern region of Ireland. A total of 2000 vaginal swabs collected in two periods in 2004 and 2006 were examined and revealed a GBS carriage rate of 16·1%. Serotyping of isolates showed that serotypes Ia, II, III, IV, and V were the most prevalent. A high prevalence of serotype IV was found, increasing from 7·6% to 15·2% between 2004 and 2006. Random amplified polymorphic DNA analysis demonstrated considerable genetic heterogeneity in the serotype IV isolates. This serotype should be considered for inclusion in potential vaccines for use in Ireland.

  16. Comparative genomic analysis of Vibrio parahaemolyticus: serotype conversion and virulence

    Gil Ana I


    Full Text Available Abstract Background Vibrio parahaemolyticus is a common cause of foodborne disease. Beginning in 1996, a more virulent strain having serotype O3:K6 caused major outbreaks in India and other parts of the world, resulting in the emergence of a pandemic. Other serovariants of this strain emerged during its dissemination and together with the original O3:K6 were termed strains of the pandemic clone. Two genomes, one of this virulent strain and one pre-pandemic strain have been sequenced. We sequenced four additional genomes of V. parahaemolyticus in this study that were isolated from different geographical regions and time points. Comparative genomic analyses of six strains of V. parahaemolyticus isolated from Asia and Peru were performed in order to advance knowledge concerning the evolution of V. parahaemolyticus; specifically, the genetic changes contributing to serotype conversion and virulence. Two pre-pandemic strains and three pandemic strains, isolated from different geographical regions, were serotype O3:K6 and either toxin profiles (tdh+, trh- or (tdh-, trh+. The sixth pandemic strain sequenced in this study was serotype O4:K68. Results Genomic analyses revealed that the trh+ and tdh+ strains had different types of pathogenicity islands and mobile elements as well as major structural differences between the tdh pathogenicity islands of the pre-pandemic and pandemic strains. In addition, the results of single nucleotide polymorphism (SNP analysis showed that 94% of the SNPs between O3:K6 and O4:K68 pandemic isolates were within a 141 kb region surrounding the O- and K-antigen-encoding gene clusters. The "core" genes of V. parahaemolyticus were also compared to those of V. cholerae and V. vulnificus, in order to delineate differences between these three pathogenic species. Approximately one-half (49-59% of each species' core genes were conserved in all three species, and 14-24% of the core genes were species-specific and in different

  17. Bacterial load of pneumococcal serotypes correlates with their prevalence and multiple serotypes is associated with acute respiratory infections among children less than 5 years of age.

    Bhim Gopal Dhoubhadel

    Full Text Available BACKGROUND: Among pneumococcal serotypes, some serotypes are more prevalent in the nasopharynx than others; determining factors for higher prevalence remain to be fully explored. As non-vaccine serotypes have emerged after the introduction of 7-valent conjugate vaccines, study of serotype specific epidemiology is in need. When two or more serotypes co-colonize, they evolve rapidly to defend host's immune responses; however, a clear association of co-colonization with a clinical outcome is lacking. METHODS: Children less than 5 years old who were admitted to hospital due to acute respiratory infections (ARI (n = 595 and healthy children (n = 350 were recruited. Carriage of pneumococcus was determined by culture and lytA PCR in the nasopharyngeal samples. Serotype/serogroup detection and its quantification were done by the nanofluidic real time PCR system. Spearman's correlation and logistic regression were used to examine a correlation of serotype/serogroup specific bacterial load with its prevalence and an association of co-colonization with ARI respectively. RESULTS: Serotype/serogroup specific bacterial load was correlated with its prevalence, both in ARI cases (Spearman's rho = 0.44, n = 186; P<0.0001 and healthy children (Spearman's rho = 0.41, n = 115; P<0.0001. The prevalence of multiple serotypes was more common in ARI cases than in healthy children (18.5% vs 7.1%; aOR 2.92, 95% CI: 1.27-6.71; P = 0.01. The dominant serotype in the co-colonization had a 2 log10 higher bacterial load than the subdominant serotype, both in ARI cases (P<0.001 and healthy children (P<0.05. CONCLUSIONS: High bacterial load in the nasopharynx may help transmit pneumococci among hosts, and increase the chance of successful acquisition and colonization. Co-colonization of multiple serotypes of pneumococci is linked with ARI, which infers the interactions of multiple serotypes may increase their pathogenicity; however, they may compete

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

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


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

  19. Evaluation of Serotype Prediction by cpsA-cpsB Gene Polymorphism in Streptococcus pneumoniae

    Lawrence, Elliot R.; Arias, Cesar A.; Duke, Brigid; Beste, Dani; Broughton, Karen; Efstratiou, Androulla; George, Robert C.; Hall, Lucinda M. C.


    New pneumococcal conjugate vaccines covering a limited number of serotypes are likely to come into widespread use over the next few years. It is unknown what effect this will have on the relative importance of different serotypes as causes of pneumococcal infection. Hence, it will be important to monitor serotype prevalence before, during, and after the introduction of new vaccines. We have investigated the ability of a PCR method based on polymorphisms in two genes common to the different ca...

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

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


    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...

  1. Abscesos pleurales producidos por Staphylococcus aureus y Streptococcus spp. beta-hemolítico como secuela de pleuroneumonía en un caballo: Reporte de un caso Abscessos pleurais causados por Staphylococcus aureus e Streptococcus spp. beta-hemolítico no rescaldo de pleuropneumonia em um cavalo.: Relato de um caso Pleural abscesses produced by Staphylococcus aureus and beta-haemolytic Streptococcus spp. as a sequel of pleuropneumonia in a horse: A case report

    Jorge U Carmona


    Full Text Available Los abscesos pleurales son una complicación frecuente en caballos con pleuroneumonía. Su tratamiento resulta difícil, requiere tiempo y dinero. Un caballo criollo argentino, castrado, de 13 años de edad presentó una pleuroneumonía grave que posteriormente se complicó con abscesos pleurales. El uso de anti-inflamatorios no esteroidales (fenilbutazona, una antibioterapia agresiva (ceftiofur, ampicilina y gentamicina, el drenaje eco-guiado y el lavado de los abscesos con una solución de yodopovidona al 0.05% permitió la recuperación exitosa del paciente de este reporte.Os abscessos pleurais são uma complicação comum em cavalos com pleuropneumonia. Seu tratamento é difícil e requer tempo e dinheiro. Um cavalo crioulo argentino, castrado, 13 anos de idade apresentou uma grave pleuropneumonia que posteriormente se tornou complicado com abscesso pleural. O uso de não-esteróides antiinflamatórios (fenilbutazona, uma agressiva terapia antibiótica (cetfiofur, ampicilina e gentamicina, eu drenagem eco-guiado e da lavagem dos abscessos com uma solução de yodopovidona de 0.05%, permitiu o sucesso da recuperação do paciente deste relato de caso.Pleural abscesses are a frequent complication in horses with pleuropneumonia. Their treatment is not easy, and requires time and it is expensive. A 13 years old creole argentine gelding presented a severe pleuropneumonia subsequently complicated with pleural abscess. The use of nonsteroidal antiinflammatory drugs (fenylbutazone, an aggressive antibiotherapy (ceftiofur, ampicillin, and gentamicin, echoguide drainage and lavage with yodopovidone to 0.05% of the abscesses permitted the successful recuperation of the patient of this case report.

  2. Development of a TaqMan Array Card for Pneumococcal Serotyping on Isolates and Nasopharyngeal Samples.

    Pholwat, Suporn; Sakai, Fuminori; Turner, Paul; Vidal, Jorge E; Houpt, Eric R


    Streptococcus pneumoniae is both a commensal and a major pathogen that causes invasive disease in people of all ages. The introduction of serotype-specific pneumococcal vaccines has reduced the burden of disease but has also led to replacement with new strains; thus, serotyping remains important for vaccine-related disease surveillance. Conventional serotyping methods are laborious and expensive. We developed an easy-to-perform genotypic TaqMan array card (TAC) to identify S. pneumoniae strains, including lytA-based sequences, and 53 sequence-specific PCRs to identify 74 serotypes/serogroups covering all current vaccine types as well as prevalent nonvaccine types. The TAC method was evaluated on 146 clinical S. pneumoniae isolates and 13 nonpneumococcal species that naturally inhabit the upper respiratory tract and yielded 97% (142/146) sensitivity and 100% (13/13) specificity versus results of standard Quellung serotyping. The calculated limit of detection was 20 to 200 fg (∼8 to 84 genome equivalents) per reaction. On 23 blinded nasopharyngeal specimens that were pneumococcus culture positive, the TAC pan-pneumococcus lytA assay was positive in 21 (91% sensitivity versus culture). On TAC lytA-positive specimens, a serotype result was obtained on 86%, and the result was 95% accurate versus the subsequent culture's Quellung result. TAC also detected mixed serotypes in two specimens where Quellung detected only the predominant serotype. This TAC method yields fast and comprehensive serotyping compared to the standard method and may be useful on direct specimens. PMID:27170020

  3. Around the World in 1,475 Salmonella Geo-serotypes

    Le Hello, Simon; de Jong, Birgitta; Rolfhamre, Per; Faensen, Daniel; Weill, François-Xavier; Giesecke, Johan


    It’s easy to remember Salmonella serotypes names, isn’t it? Surely, this is because the naming system of Salmonella serotypes is by far the most scientist friendly. Traditionally, most Salmonella serotypes have been named after geographic locations. We decided to explore the geographic locations to which Salmonella serotypes refer and describe some unexpected twists in the naming scheme. We found that 93% (n = 1,475) of the 1,585 serotypes could be categorized as geo-serotypes; that is, the name refers to a geographic location. The 3 countries with the most geo-serotypes are Germany, the United Kingdom, and the United States. Other serotype names refer to the name of a person, animal, tribe, or food item or are a composite of symptoms and host. The Salmonella serotypes naming scheme has had a valuable effect on public health microbiology, and in the current era of fast development of whole-genome sequencing, it should remain a reference.

  4. Serological characterization of Streptococcus mutans serotype polysaccharide g and its different molecular weight forms.

    Takada, K; Wyszomirska, J; Shiota, T


    The serotype polysaccharide g from Streptococcus mutans 6715 was found to cross-react with serotype polysaccharide a from S. mutans HS6 and serotype polysaccharide d from S. mutans B13. Double immunodiffusion experiments indicated that the serotype polysaccharide g consisted of the following: (i) the type-specific g site; (ii) a cross-reactive site g-a that was in common with polysaccharide a; (iii) a cross-reactive site g-d that was in common with polysaccharide d; and (iv) a cross-reactive ...

  5. Serotype Specific Invasive Capacity and Persistent Reduction in Invasive Pneumococcal Disease

    Yildirim, Inci; Hanage, William P.; Lipsitch, Marc; Shea, Kimberly M.; Stevenson, Abbie; Finkelstein, Jonathan; Huang, Susan S.; Lee, Grace M.; Kleinman, Ken; Pelton, SI


    Defining the propensity of Streptoccocus pneumoniae (SP) serotypes to invade sterile body sites following nasopharyngeal (NP) acquisition has the potential to inform about how much invasive pneumococcal disease (IPD) may occur in a typical population with a given distribution of carriage serotypes. Data from enhanced surveillance for IPD in Massachusetts children ≤7 years in 2003/04, 2006/07 and 2008/09 seasons and surveillance of SP NP carriage during the corresponding respiratory seasons in 16 Massachusetts communities in 2003/04 and 8 of the 16 communities in both 2006/07 and 2008/09 were used to compute a serotype specific “invasive capacity (IC)” by dividing the incidence of IPD due to serotype x by the carriage prevalence of that same serotype in children of the same age. A total of 206 IPD and 806 NP isolates of SP were collected during the study period. An approximate 50-fold variation in the point estimates between the serotypes having the highest (18C, 33F, 7F, 19A, 3 and 22F) and lowest (6C, 23A, 35F, 11A, 35B, 19F, 15A, and 15BC) IC was observed. Point estimates of IC for most of the common serotypes currently colonizing children in Massachusetts were low and likely explain the continued reduction in IPD from the pre-PCV era in the absence of specific protection against these serotypes. Invasive capacity differs among serotypes and as new pneumococcal conjugate vaccines are introduced, ongoing surveillance will be essential to monitor whether serotypes with high invasive capacity emerge (e.g. 33F, 22F) as successful colonizers resulting in increased IPD incidence due to replacement serotypes. PMID:21029807

  6. Phylogeography of Dengue Virus Serotype 4, Brazil, 2010-2011

    Nunes, Marcio Roberto Teixeira; Faria, Nuno Rodrigues; Vasconcelos, Helena Baldez; Medeiros, Daniele Barbosa de Almeida; Silva de Lima, Clayton Pereira; Carvalho, Valéria Lima; Pinto da Silva, Eliana Vieira; Cardoso, Jedson Ferreira; Sousa, Edivaldo Costa; Nunes, Keley Nascimento Barbosa; Rodrigues, Sueli Guerreiro; Abecasis, Ana Barroso; Suchard, Marc A.; Lemey, Philippe; Vasconcelos, Pedro Fernando da Costa


    Dengue virus serotype 4 (DENV-4) reemerged in Roraima State, Brazil, 28 years after it was last detected in the country in 1982. To study the origin and evolution of this reemergence, full-length sequences were obtained for 16 DENV-4 isolates from northern (Roraima, Amazonas, Pará States) and northeastern (Bahia State) Brazil during the 2010 and 2011 dengue virus seasons and for an isolate from the 1982 epidemic in Roraima. Spatiotemporal dynamics of DENV-4 introductions in Brazil were applie...

  7. Food poisoning due to Salmonella enterica serotype Weltevreden in Mangalore

    Antony B


    Full Text Available An outbreak of food poisoning due to Salmonella enterica serotype Weltevreden ( S.weltevreden involving 34 students has been reported from a tertiary care hospital in Mangalore. The symptoms developed 8-10 hours, after consuming a non- vegetarian dish, probably fish, from an outside caterer. The identity of the organism was confirmed at Central Research Institute, Kasauli. This report emphasizes the geographical distribution of this organism in the Coastal Karnataka region. S.Weltevreden may be overlooked due to the biochemical similarity to S. Paratyphi B & S. Typhimurium.

  8. Comparative sequence analyses of the neurotoxin complex genes in Clostridium botulinum serotypes A, B, E, and F

    Ajay K. Singh


    Full Text Available Neurotoxin complex (NTC genes are arranged in two known hemagglutinin (HA and open reading frame X (ORFX clusters. NTC genes have been analyzed in four serotypes A, B, E and F of Clostridium botulinum causing human botulism. Analysis of amino acid sequences of NT genes demonstrated significant differences among subtypes and four serotypes. Phylogram tree of NT genes reveals that serotypes A1 and B1 are much closer compared to serotype E1 and F1. However, non-toxic non-hemagglutinin (NTNH gene is highly conserved among four serotypes. Analysis of phylogram tree of NTNH gene reveals that serotypes A and F are more closely related compared to serotype B and E. Additionally, sequences of HAs and ORFX genes are very divergent but these genes are specific in subtypes and serotypes of Clostridium botulinum. Information derived from sequence analyses of NTC has direct implication in development of detection tools and therapeutic countermeasures for botulism.

  9. Generation of serotype 1/serotype 2 reassortant viruses of the infectious bursal disease virus and their investigation in vitro and in vivo.

    Zierenberg, Kati; Raue, Rüdiger; Nieper, Hermann; Islam, Md Rafiqul; Eterradossi, Nicolas; Toquin, Didier; Müller, Hermann


    Infectious bursal disease virus (IBDV) is the causative agent of acute or immunosuppressive disease in chickens. Serotype 1 strains are pathogenic whereas serotype 2 strains neither cause disease nor protect against infection with the serotype 1 strains. The target organ of serotype 1 strains is the bursa Fabricii (BF). The molecular determinants of this tropism, and therefore pathogenicity, are poorly understood. IBDV is a non-enveloped icosahedral virus particle of 60 nm in diameter, which contains two genome segments of double-stranded RNA. Here, the generation of interserotypic reassortants using the reverse genetics approach is reported. The results of in vitro and in vivo investigations show that genome segment A determines the bursa tropism of IBDV, whereas segment B is involved in the efficiency of viral replication; they further indicate the significance of the interaction of the polymerase (segment B) with the structural protein VP3 (segment A) or the viral genome for efficient virus formation and replication. PMID:15325078

  10. Modified indirect immunofluorescence test for serotyping large numbers of Ureaplasma urealyticum clinical isolates.

    Naessens, A; Lauwers, S


    A technical modification of the indirect immunofluorescence test for serotyping Ureaplasma urealyticum clinical isolates is described. The use of a tissue culture plate filled with ureaplasma agar made the serotyping easier to perform and proved to be very handy, especially for performance of large series.

  11. A monoclonal antibody based capture ELISA for botulinum neurotoxin serotype B: toxin detection in food

    Botulism is a serious foodborne neuroparalyic disease caused by botulinum neurotoxin (BoNT) produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A-H) have been described. The majority of human cases of botulism are caused by serotypes A and B followed by E and F. We repo...

  12. Complete Genome Sequence of Aggregatibacter actinomycetemcomitans Serotype g Strain NUM4039 (JCM 30399)

    Saito, Masanori; Hirasawa, Masaaki; Kuwahara, Noriko; Okada, Tamami; Umezawa, Koji; Kobayashi, Taira; Okamoto, Masaaki; Naito, Mariko; Hirasawa, Masatomo


    Aggregatibacter actinomycetemcomitans is considered to be a major etiological agent of aggressive periodontitis and includes serotype a to g strains. We herein report the first complete genome sequence of A. actinomycetemcomitans serotype g strain NUM4039. The genome is 2,382,853 bp in length with a G+C content of 44.34%. PMID:26988057

  13. Unravelling Selection Shifts Among Foot-and-Mouth Disease Virus (FMDV Serotypes

    Mario A. Fares


    Full Text Available FMDV virus has been increasingly recognised as the most economically severe animal virus with a remarkable degree of antigenic diversity. Using an integrative evolutionary and computational approach we have compelling evidence for heterogeneity in the selection forces shaping the evolution of the seven different FMDV serotypes. Our results show that positive Darwinian selection has governed the evolution of the major antigenic regions of serotypes A, Asia1, O, SAT1 and SAT2, but not C or SAT3. Co-evolution between sites from antigenic regions under positive selection pinpoints their functional communication to generate immune-escape mutants while maintaining their ability to recognise the host-cell receptors. Neural network and functional divergence analyses strongly point to selection shifts between the different serotypes. Our results suggest that, unlike African FMDV serotypes, serotypes with wide geographical distribution have accumulated compensatory mutations as a strategy to ameliorate the effect of slightly deleterious mutations fixed by genetic drift. This strategy may have provided the virus by a flexibility to generate immune-escape mutants and yet recognise host-cell receptors. African serotypes presented no evidence for compensatory mutations. Our results support heterogeneous selective constraints affecting the different serotypes. This points to the possible accelerated rates of evolution diverging serotypes sharing geographical locations as to ameliorate the competition for the host.

  14. Experimental infection of white-tailed deer with bluetongue virus serotype 8

    Drolet, B.S.; Reister, L.M.; Mecham, J.O.; Wilson, W.C.; Nol, P.; Vercauteren, K.C.; Rijn, van P.A.; Bowen, R.A.


    Bluetongue (BT) is an insect-transmitted, economically important disease of domestic and wild ruminants. Although only five of the 26 reported bluetongue virus (BTV) serotypes are considered endemic to the USA, 10 exotic serotypes have been isolated primarily in the southeastern region of the countr

  15. Susceptibility of North American white-tailed deer to the Netherlands strain of BTV serotype 8

    World-wide there are at least 24 serotypes of bluetongue virus (BTV), a complex non-enveloped virus in the family Reoviridae, genus Orbivirus. Bluetongue (BT) is an arthropod-borne disease of cattle, sheep, goats, and deer and is transmitted by Culicoides biting midges. In 2006, bluetongue serotype ...

  16. Two distinct outer membrane serotype subcomplexes of Neisseria meningitidis serogroup A.

    Bläsius, R; Strittmatter, W; Crowe, B.; Achtman, M.


    A 350-kilodalton serotype outer membrane complex containing the class 1, 3, and 4 outer membrane proteins was isolated from serogroup A Neisseria meningitidis. Partial denaturation yielded two serotype subcomplexes containing the class 3 and 1 proteins (85 kilodaltons) and the class 3 and 4 proteins (94 kilodaltons), respectively.

  17. Draft Genome Sequences of Streptococcus agalactiae Serotype Ia and III Isolates from Tilapia Farms in Thailand.

    Areechon, Nontawith; Kannika, Korntip; Hirono, Ikuo; Kondo, Hidehiro; Unajak, Sasimanas


    Streptococcus agalactiaeserotypes Ia and III were isolated from infected tilapia in cage and pond culture farms in Thailand during 2012 to 2014, in which pathogenicity analysis demonstrated that serotype III showed higher virulence than serotype Ia. Here, we report the draft genome sequencing of piscineS. agalactiaeserotypes Ia and III. PMID:27013037

  18. Pneumococcal serotypes and mortality following invasive pneumococcal disease: a population-based cohort study

    Harboe, Zitta B; Thomsen, Reimar W; Riis, Anders;


    BACKGROUND: Pneumococcal disease is a leading cause of morbidity and mortality worldwide. The aim of this study was to investigate the association between specific pneumococcal serotypes and mortality from invasive pneumococcal disease (IPD). METHODS AND FINDINGS: In a nationwide population......-based cohort study of IPD in Denmark during 1977-2007, 30-d mortality associated with pneumococcal serotypes was examined by multivariate logistic regression analysis after controlling for potential confounders. A total of 18,858 IPD patients were included. Overall 30-d mortality was 18%, and 3% in children...... compared with serotype 1 (all: adjusted odds ratio >or=3, p<0.001). In children younger than 5 y, associations between serotypes and mortality were different than in adults but statistical precision was limited because of low overall childhood-related mortality. CONCLUSIONS: Specific pneumococcal serotypes...

  19. Recurrent Invasive Pneumococcal Disease Serotype 12F in a Vaccinated Splenectomized Patient

    Blaabjerg, Anne Katrine; Schumacher, Anna Holst; Kantsø, Bjørn;


    This is the first case report of recurrent invasive pneumococcal disease (IPD), specifically, due to serotype 12F. The patient described here was vaccinated with the 23-valent pneumococcal polysaccharide vaccine (PPV23) due to previous splenectomy, and an anti-pneumococcal IgG test concluded that...... she had responded sufficiently to vaccination. Still, she had a fulminate recurrent infection with PPV23 serotype 12F. We investigated the anti-pneumococcal IgG test, and it turned out that it is based on the geometric mean value of only 12 of the serotypes included in PPV23; 12F is none of them. The...... reason is that there are no titer cut-offs available for 11 of the PPV23 serotypes, including 12F, neither nationally nor internationally. Yet, this is not specified in the answer to the clinicians. This case illustrates the need for titer cut-offs for the remaining pneumococcal serotypes in available...

  20. Temporal trends in invasive pneumococcal disease and pneumococcal serotypes over 7 decades

    Harboe, Zitta B; Benfield, Thomas L; Valentiner-Branth, Palle;


    BACKGROUND: Pneumococcal infections have historically played a major role in terms of morbidity and mortality. We explored historical trends of invasive pneumococcal disease (IPD) and pneumococcal serotypes in a population exposed to limited antibiotic selective pressure and conjugate pneumococcal...... vaccination (PCV). METHODS: Retrospective cohort study based on nationwide laboratory surveillance data on IPD collected uninterruptedly in Denmark during 1938-2007. Changes in the reported incidence and trends of pneumococcal serotypes were explored using nonlinear regression analysis. RESULTS: There were 25...... serotype 19A increased before introduction of PCV. Between 1993 and 2007, the level of resistance to macrolides and beta-lactams was 6%. CONCLUSIONS: The epidemiology of IPD and single serotypes has constantly changed over the past 7 decades. PCV serotypes appeared to dominate the pneumococcal population....

  1. Antibodies to Staphylococcus aureus Serotype 8 Capsular Polysaccharide React with and Protect against Serotype 5 and 8 Isolates

    Park, Saeyoung; Gerber, Sabina; Lee, Jean C.


    Most Staphylococcus aureus isolates produce either a serotype 5 (CP5) or 8 (CP8) capsular polysaccharide, and the CP antigens are targets for vaccine development. Since CP5 and CP8 have similar trisaccharide repeating units, it is important to identify an epitope shared by both CP5 and CP8. To characterize cross-reactivity between CP5 and CP8, the immunogenicity of CP5 and CP8 conjugate vaccines in mice and rabbits was evaluated by serological assays. Immune sera were also tested for function...

  2. Molecular characterization and analysis of high-level multidrug-resistance of Shigella flexneri serotype 4s strains from China

    Yang, Chaojie; Li, Peng; Zhang, Xiujuan; Ma, Qiuxia; Cui, Xianyan; Li, Hao; Liu, Hongbo; Wang, Jian; Xie, Jing; Wu, Fuli; Sheng, Chunyu; Du, Xinying; Qi, Lihua; Su, Wenli; Jia, Leili; Xu, Xuebin; Zhao, Jiayong; Xia, Shengli; Zhou, Na; Ma, Hui; Qiu, Shaofu; Song, Hongbin


    To conduct the first comprehensive analysis of Shigella flexneri serotype 4s, a novel serotype found in 2010, we identified 24 serotype 4s isolates from 1973 shigellosis cases in China (2002–2014). The isolates were characterized by single nucleotide polymorphism (SNP) phylogenetic analysis, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) to determine their genetic relatedness, and analysed further for their antimicrobial susceptibilities and antimicrobial resistance determinants. The PFGE and SNP phylogenetic analyses suggest that S. flexneri serotype 4s strains are derived from multiple serotypes, including two predominant serotypes in China: serotype X variant and serotype II. Three new sequence types were identified by MLST. All isolates were resistant to ticarcillin, ampicillin and tetracycline, with high-level resistance to third-generation cephalosporins. Notably, all the isolates were multidrug resistant (MDR), with the highest levels of resistance observed for eight antimicrobials classes. Most isolates contain various antimicrobial resistance determinants. In conclusion, we found that serotype 4s isolates have multiple evolutionary sources, diverse biochemical characteristics and genomes, and highly prevalent multidrug resistance and antimicrobial-resistant determinants. With few clinical treatment options, continuous monitoring and timely intervention against this emerging MDR serotype is essential. The possibility that serotype 4s will become the next predominant serotype exists. PMID:27374009

  3. Adrenal gland infection by serotype 5 adenovirus requires coagulation factors.

    Lucile Tran

    Full Text Available Recombinant, replication-deficient serotype 5 adenovirus infects the liver upon in vivo, systemic injection in rodents. This infection requires the binding of factor X to the capsid of this adenovirus. Another organ, the adrenal gland is also infected upon systemic administration of Ad, however, whether this infection is dependent on the cocksackie adenovirus receptor (CAR or depends on the binding of factor X to the viral capsid remained to be determined. In the present work, we have used a pharmacological agent (warfarin as well as recombinant adenoviruses lacking the binding site of Factor X to elucidate this mechanism in mice. We demonstrate that, as observed in the liver, adenovirus infection of the adrenal glands in vivo requires Factor X. Considering that the level of transduction of the adrenal glands is well-below that of the liver and that capsid-modified adenoviruses are unlikely to selectively infect the adrenal glands, we have used single-photon emission computed tomography (SPECT imaging of gene expression to determine whether local virus administration (direct injection in the kidney could increase gene transfer to the adrenal glands. We demonstrate that direct injection of the virus in the kidney increases gene transfer in the adrenal gland but liver transduction remains important. These observations strongly suggest that serotype 5 adenovirus uses a similar mechanism to infect liver and adrenal gland and that selective transgene expression in the latter is more likely to be achieved through transcriptional targeting.

  4. Prevalence and Characteristics of Salmonella Serotypes Isolated from Fresh Produce Marketed in the United States.

    Reddy, Shanker P; Wang, Hua; Adams, Jennifer K; Feng, Peter C H


    Salmonella continues to rank as one of the most costly foodborne pathogens, and more illnesses are now associated with the consumption of fresh produce. The U.S. Department of Agriculture Microbiological Data Program (MDP) sampled select commodities of fresh fruit and vegetables and tested them for Salmonella, pathogenic Escherichia coli, and Listeria. The Salmonella strains isolated were further characterized by serotype, antimicrobial resistance, and pulsed-field gel electrophoresis profile. This article summarizes the Salmonella data collected by the MDP between 2002 and 2012. The results show that the rates of Salmonella prevalence ranged from absent to 0.34% in cilantro. A total of 152 isolates consisting of over 50 different serotypes were isolated from the various produce types, and the top five were Salmonella enterica serotype Cubana, S. enterica subspecies arizonae (subsp. IIIa) and diarizonae (subsp. IIIb), and S. enterica serotypes Newport, Javiana, and Infantis. Among these, Salmonella serotypes Newport and Javiana are also listed among the top five Salmonella serotypes that caused most foodborne outbreaks. Other serotypes that are frequent causes of infection, such as S. enterica serotypes Typhimurium and Enteritidis, were also found in fresh produce but were not prevalent. About 25% of the MDP samples were imported produce, including 65% of green onions, 44% of tomatoes, 42% of hot peppers, and 41% of cantaloupes. However, imported produce did not show higher numbers of Salmonella-positive samples, and in some products, like cilantro, all of the Salmonella isolates were from domestic samples. About 6.5% of the Salmonella isolates were resistant to the antimicrobial compounds tested, but no single commodity or serotype was found to be the most common carrier of resistant strains or of resistance. The pulsed-field gel electrophoresis profiles of the produce isolates showed similarities with Salmonella isolates from meat samples and from outbreaks, but

  5. Serotype distribution of Streptococcus pneumoniae in children with invasive diseases in Turkey: 2008-2014.

    Ceyhan, Mehmet; Ozsurekci, Yasemin; Gürler, Nezahat; Öksüz, Lütfiye; Aydemir, Sohret; Ozkan, Sengul; Yuksekkaya, Serife; Keser Emiroglu, Melike; Gültekin, Meral; Yaman, Akgün; Kiremitci, Abdurrahman; Yanık, Keramettin; Karli, Arzu; Ozcinar, Hatice; Aydin, Faruk; Bayramoglu, Gulcin; Zer, Yasemin; Gulay, Zeynep; Gayyurhan, Efgan Dogan; Gül, Mustafa; Özakın, Cüneyt; Güdücüoğlu, Hüseyin; Perçin, Duygu; Akpolat, Nezahat; Ozturk, Candan; Camcıoğlu, Yıldız; Karadağ Öncel, Eda; Çelik, Melda; Şanal, Laser; Uslu, Hakan


    Successful vaccination policies for protection from invasive pneumococcal diseases (IPD) dependent on determination of the exact serotype distribution in each country. We aimed to identify serotypes of pneumococcal strains causing IPD in children in Turkey and emphasize the change in the serotypes before and after vaccination with 7-valent pneumococcal conjugate vaccine (PCV-7) was included and PCV-13 was newly changed in Turkish National Immunization Program. Streptococcus pneumoniae strains were isolated at 22 different hospitals of Turkey, which provide healthcare services to approximately 65% of the Turkish population. Of the 335 diagnosed cases with S. pneumoniae over the whole period of 2008-2014, the most common vaccine serotypes were 19F (15.8%), 6B (5.9%), 14 (5.9%), and 3 (5.9%). During the first 5 y of age, which is the target population for vaccination, the potential serotype coverage ranged from 57.5 % to 36.8%, from 65.0% to 44.7%, and from 77.4% to 60.5% for PCV-7, PCV-10, and PCV-13 in 2008-2014, respectively. The ratio of non-vaccine serotypes was 27.2% in 2008-2010 whereas was 37.6% in 2011-2014 (p=0.045). S. penumoniae serotypes was less non-susceptible to penicillin as compared to our previous results (33.7 vs 16.5 %, p=0.001). The reduction of those serotype coverage in years may be attributed to increasing vaccinated children in Turkey and the increasing non-vaccine serotype may be explained by serotype replacement. Our ongoing IPD surveillance is a significant source of information for the decision-making processes on pneumococcal vaccination. PMID:26325175

  6. Generation of nalidixic acid-resistant strains and signature-tagged mutants of Actinobacillus pleuropneumoniae%胸膜肺炎放线杆菌萘啶酸抗性菌株的选育和信号标签突变株的构建

    商霖; 李薇; 李良军; 黎璐; 张四化; 李婷婷; 李耀坤; 刘磊; 郭志伟; 周锐; 陈焕春


    胸膜肺炎放线杆菌(APP)是重要的猪呼吸道病原菌,给世界养猪业造成严重的经济损失.信号标签突变(STM)技术是在宿主动物体内鉴定病原菌毒力因子的高通量方法.通过体外传代选育出APP血清1型和3型萘啶酸抗性菌株,再以萘啶酸抗性菌株为受体菌,以携带mini-Tn10的标签质粒(pLOF/TAG1-48)的E.coli CC118 λ pir或S17-1λpir为供体菌,在或不在E.coli DH5α(pRK2073)的辅助下,进行三亲本或两亲本接合,通过抗性筛选、PCR和Southern杂交鉴定转座突变株.结果表明:体外萘啶酸加压传代很容易选育出萘啶酸抗性APP菌株,该抗性的产生与DNA促旋酶A亚基基因gyrA的突变有关.在APP与E. coli接合实验中,两亲本接合比三亲本接合操作更简单,效率也较高;APP不同菌株在接合和转座效率上存在很大差异,血清1型菌株高于血清3型菌株,3型标准菌株高于地方分离株JL03-R.本研究为APP STM突变体库的构建与毒力基因的鉴定奠定了基础.

  7. VP7 from African horse sickness virus serotype 9 protects mice against a lethal, heterologous serotype challenge.

    Wade-Evans, A M; Pullen, L; Hamblin, C; O'Hara, R S; Burroughs, J N; Mertens, P P


    An established mouse model system was used to evaluate the effectiveness of the major outer core protein VP7 of African horse sickness virus (AHSV) serotype 9 as a subunit vaccine. Balb C mice were immunised with VP7 crystals purified from AHSV infected BHK cells. In groups of mice, each of which was immunised with > or = 1.5 micrograms of the protein in Freund's adjuvant, > or = 80% of mice survived challenge with a virulent strain of a heterologous AHSV serotype (AHSV 7), that killed > or = 80% of the mice in the uninoculated control groups. This level of protection was significantly greater than that observed in mice inoculated with equivalent amounts of either denatured VP7 (50% survival), or GST/VP7 fusion protein (50-70% survival), or which were vaccinated with AHSV 9 (40-50% survival). The VP7 protein folding, or its assembly into crystals, are thought to play some role in the effectiveness of the protective response observed. Titres of circulating antibodies against AHSV VP7 were determined by competitive ELISA but did not appear to correlate with the levels of protection observed. Passive transfer of these antibodies to syngeneic recipients also failed to protect Balb C mice from the AHSV 7 challenge. The observed protection is therefore unlikely to be due to an antibody mediated immune response. PMID:9785508

  8. Development of a Novel Cocktail Enzyme-Linked Immunosorbent Assay and a Field-Applicable Lateral-Flow Rapid Test for Diagnosis of Contagious Bovine Pleuropneumonia.

    Heller, Martin; Gicheru, Nimmo; Tjipura-Zaire, Georgina; Muriuki, Cecilia; Yu, Mingyan; Botelho, Ana; Naessens, Jan; Jores, Joerg; Liljander, Anne


    Contagious bovine pleuropneumonia (CBPP) is a severe respiratory disease that is widespread in sub-Saharan Africa. It is caused by Mycoplasma mycoides subsp. mycoides, a bacterium belonging to the Mycoplasma mycoides cluster. In the absence of an efficient CBPP vaccine, improved and easy-to-use diagnostic assays for recurrent testing combined with isolation and treatment of positive animals represent an option for CBPP control in Africa. Here we describe the comprehensive screening of 17 immunogenic Mycoplasma mycoides subsp. mycoides proteins using well-characterized bovine sera for the development of a novel cocktail enzyme-linked immunosorbent assay (ELISA) for laboratory use. Two recombinant Mycoplasma immunogens, MSC_0136 and MSC_0636, were used to set up a standardized cocktail ELISA protocol. According to the results from more than 100 serum samples tested, the sensitivity and specificity of the novel cocktail ELISA were 85.6% and 96.4%, respectively, with an overall diagnostic accuracy comparable to that of the Office International des Epizooties (OIE)-prescribed serological assays. In addition, we provide a proof of principle for a field-applicable, easy-to-use commercially produced prototype lateral-flow test for rapid (<30-min) diagnosis of CBPP. PMID:27053669

  9. Experimental Contagious Caprine Pleuropneumonia: A Long Term Study on the Course of Infection and Pathology in a Flock of Goats Infected with Mycoplasma capricolum subsp. capripneumoniae

    Bölske G


    Full Text Available Contagious caprine pleuropneumonia (CCPP is a major threat to goat farming in parts of Africa and Asia. It classically causes acute high morbidity and mortality early in infection, but little is known of its long term epizootiology and course. In this study, 10 goats were inoculated with Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae and then mixed with 15 goats for contact transmission. The disease course was monitored in each goat for 56–105 days, whereafter the goats were killed and necropsied. Varying features signifying infection occurred in altogether 17 goats (7 inoculated, 10 in-contact. Clinical signs were severe in 8 goats but no fatalities occurred. Only 6 goats had serum antibody titres against M. capripneumoniae in ELISA. Fourteen goats (5 inoculated, 9 in-contact had chronic pleuropulmonary lesions compatible with CCPP at necropsy and 7 of those showed M. capripneumoniae antigen in the lung by immunohistochemistry. Neither cultivation nor PCR tests were positive for the agent in any goat. The results indicate that the clinical course of CCPP in a flock may be comparatively mild, M. capripneumoniae-associated lung lesions may be present at a late stage of infection, and chronic infection may occur without a significant serological response.

  10. Determination of pneumococcal serotypes in meningitis cases in Niger, 2003-2011.

    Jean-Marc Collard

    Full Text Available BACKGROUND: The epidemiology of pneumococcal meningitis in the African 'meningitis belt' is poorly studied. In order to ensure an effective vaccination strategy and post-vaccination surveillance, we examined the serotype distribution patterns of pneumococcal meningitis in Niger over the period 2003-2011. METHODS: Cerebrospinal fluid (CSF samples were collected from different health facilities throughout Niger in the frame of the national microbiological surveillance of meningitis. Determination of the serotype of CSF positive for pneumococci was performed using a sequential multiplex PCR method (SM-PCR adapted with a national algorithm in which 32 different serotypes were covered and grouped into eight consecutive PCR. RESULTS: The SM-PCR assay could predict the Sp serotype for 779 CSF (88.7%, 98 CSF (11.3% were not-typeable in our national-adapted algorithm. In total, 26 different serotypes were identified. Serotype 1 (n = 393 was the most prevalent and accounted for 45.3% of infections, followed by serogroups/serotypes 12F/(12A/(44/(46 (7.3%, 6/(6A/6B/6C/6D (5.4%, 14 (5.2%, 5 (4.6%, 23F (4.2%, 45 (3.6%, 2 (3.1%, 18/(18A/18B/18C/18F (2.9% and 17 others serotypes with a prevalence of less than 2%. The proportion of serotype 1 in infants(<2 years old represented only 4.3% of the cases affected by this serotype. In contrast, serotypes 5, 6, 14, 19A and 23F were only detected in very young children. CONCLUSIONS: The proportion of serotype 1 in the pneumococcal meningitis cases and the theoretical vaccine coverage across all age groups advocates for the introduction of a conjugate vaccine (PCV10 or 13 into the Expanded Programme on Immunization (EPI in Niger. Post-vaccine introduction surveillance supported by molecular approaches will be essential to provide a comprehensive picture of the impact of the vaccine on the burden reduction of pneumococcal meningitis and on pneumococcal serotype distribution.

  11. Occurrence of Actinobacillus actinomycetemcomitans in patients with chronic periodontitis, aggressive periodontitis, healthy subjects and children with gingivitis in two cities of the state of São Paulo, Brazil Ocorrência de Actinobacillus actinomycetemcomitans em pacientes com periodontite crônica, periodontite agressiva, pessoas saudáveis e crianças com gengivite em duas cidades do Estado de São Paulo, Brasil

    Elerson Gaetti Jardim Júnior; Joseane Maria Dias Bosco; Angélica Marquezim Lopes; Luís Fernando Landucci; Ellen Cristina Gaetti Jardim; Sílvia Rosana Soares Carneiro


    The aim of this study was to determine the frequency of isolation of Actinobacillus actinomycetemcomitans (Aa) in 100 patients with chronic periodontitis, 14 patients with aggressive periodontitis, 142 pre-school children with gingivitis and 134 periodontally healthy subjects. Samples of subgingival plaque were taken using sterilized paper points introduced into periodontal pockets or gingival crevice for 60 seconds and inoculated on TSBV agar, which was incubated under anaerobiosis at 37ºC, ...

  12. A novel high-throughput method for molecular serotyping and serotype-specific quantification of Streptococcus pneumoniae using a nanofluidic real-time PCR system.

    Dhoubhadel, Bhim Gopal; Yasunami, Michio; Yoshida, Lay-Myint; Thi, Hien Anh Nguyen; Thi, Thu Huong Vu; Thi, Thuy Ai Nguyen; Watanabe, Kiwao; Suzuki, Motoi; Morimoto, Konosuke; Dang, Duc Anh; Ariyoshi, Koya


    Serotype-specific quantification data are essential for elucidating the complex epidemiology of Streptococcus pneumoniae and evaluating pneumococcal vaccine efficacy. Various PCR-based assays have been developed to circumvent the drawback of labour-intensive and time-consuming culture-based procedures for serotype determination and quantification of pneumococcus. Here, we applied a nanofluidic real-time PCR system to establish a novel assay. Twenty-nine primer pairs, 13 of which were newly designed, were selected for the assay to cover 50 serotypes including all currently available conjugate and polysaccharide vaccine serotypes. All primer pairs were evaluated for their sensitivity, specificity, efficiency, repeatability, accuracy and reproducibility on the Fluidigm Biomark HD System, a nanofluidic real-time PCR system, by drawing standard curves with a serial dilution of purified DNA. We applied the assay to 52 nasopharyngeal swab samples from patients with pneumonia confirmed by chest X-ray to validate its accuracy. Minimum detection levels of this novel assay using the nanofluidic real-time PCR system were comparable to the conventional PCR-based assays (between 30 and 300 copies per reaction). They were specific to their targets with good repeatability (sd of copy number of 0.1), accuracy (within ±0.1 fold difference in log10 copy number) and reproducibility (sd of copy number of 0.1). When artificially mixed DNA samples consisting of multiple serotypes in various ratios were tested, all the serotypes were detected proportionally, including a minor serotype of one in 1000 copies. In the nasopharyngeal samples, the PCR system detected all the culture-positive samples and 22 out of 23 serotypes identified by the conventional method were matched with PCR results. We conclude that this novel assay, which is able to differentially quantify 29 pneumococcus groups for 45 test samples in a single run, is applicable to the large-scale epidemiological study of

  13. Serotype markers in a Streptococcus agalactiae strain collection from Zimbabwe

    Mavenyengwa R


    Full Text Available Objective: Group B streptococci (GBS from Southern African areas have been less well characterized. Our objective was to study serotype and serovariant distribution of carrier GBS strains as part of a study of the epidemiology of GBS carriage in pregnant women from Zimbabwe. Materials and Methods: We studied GBS isolated from 121 healthy pregnant women living in Harare and surrounding areas, Zimbabwe. Capsular polysaccharide (CPS testing for serotype determination and surface-anchored protein testing for serosubtype determination were done by gene-based serotyping (PCR, except for the proteins R3 and a novel protein called Z, which were detected by antibody-based methods. Results: Strains of the CPS types Ia (15.7%, Ib (11.6%, II (8.3%, III (38.8%, V (24.0% and NT (1.7% were detected along with the strain-variable proteins Cί (15.7% of isolates, Cα (19.8%, Alp1 (epsilon-22.3%, Alp3 (5.0%, R4/Rib (46.3%, R3 (27.3%, Z (27.3%, and SAR5 (28.9%, which encodes the R5 protein. Up to four of the protein genes could be possessed or the gene product expressed by one and the same isolate. A total of 32 serovariants were detected. The findings assessed by us as most important were the very low prevalence of the gene Alp3 (Alp3 - 4.9%, high prevalence of R4 (Rib - 46.2%, the proteins R3 (27.3%, Z (27.3%, and of SAR5 (R5 - 28.9%. The low prevalence of Alp3, notably in GBS type V strains, differed from findings with CPS type V GBS from non-African areas. Bacteria of the various CPS types showed distinct CPS/protein-marker associations. Conclusion: The results are of importance in relation to regional variations of GBS phenotypes and genotypes and thus, of importance in planning and research in the context of future vaccine formulations.

  14. Epidemiological analysis of pneumococcal serotype 19A in healthy children following PCV7 vaccination.

    Tóthpál, A; Laub, K; Kardos, S; Tirczka, T; Kocsis, A; VAN DER Linden, M; Dobay, O


    After the introduction of conjugate vaccines, a strong rearrangement of pneumococcal serotypes was observed globally. Probably most concerning was the emergence of serotype 19A, which has not only high invasive disease potential, but also high antibiotic resistance. In the current study we focused on the increased prevalence of serotype 19A after the PCV vaccination rate became widely used in Hungary. A total of 2262 children aged 3-6 years were screened for pneumococcus carriage using nasal swabs. Children were divided into two groups according to the vaccination rates, low level (group 1) vs. high level (group 2). While the carriage rate did not change over time (average 32·9%), the serotype distribution differed greatly in the two groups. The prevalence of serotype 19A increased >eightfold. Almost all 19A isolates had high-level macrolide resistance and elevated penicillin minimum inhibitory concentrations. Genotyping methods revealed that these new 19A isolates are different from the previously frequent Hungary19A-6 PMEN clone. Both the carriage rate and the overall penicillin and macrolide resistance remained stable over time, but while several serotypes were represented in group 1, serotype 19A alone was clearly dominant in group 2. PMID:26548594

  15. El Niño-Southern Oscillation, local weather and occurrences of dengue virus serotypes

    Huang, Xiaodong; Clements, Archie C. A.; Williams, Gail; Devine, Gregor; Tong, Shilu; Hu, Wenbiao


    Severe dengue fever is usually associated with secondary infection by a dengue virus (DENV) serotype (1 to 4) that is different to the serotype of the primary infection. Dengue outbreaks only occur following importations of DENV in Cairns, Australia. However, the majority of imported cases do not result in autochthonous transmission in Cairns. Although DENV transmission is strongly associated with the El Niño-Southern Oscillation (ENSO) climate cycle and local weather conditions, the frequency and potential risk factors of infections with the different DENV serotypes, including whether or not they differ, is unknown. This study used a classification tree model to identify the hierarchical interactions between Southern Oscillation Index (SOI), local weather factors, the presence of imported serotypes and the occurrence of the four autochthonous DENV serotypes from January 2000-December 2009 in Cairns. We found that the 12-week moving average of SOI and the 2-week moving average of maximum temperature were the most important factors influencing the variation in the weekly occurrence of the four DENV serotypes, the likelihoods of the occurrence of the four DENV serotypes may be unequal under the same environmental conditions, and occurrence may be influenced by changes in global and local environmental conditions in Cairns.

  16. Serotype distribution of Streptococcus pneumoniae causing invasive disease in the Republic of Ireland.

    Vickers, I


    The 7-valent pneumococcal conjugate vaccine (PCV7) was included in the routine infant immunization schedule in Ireland in September 2008. We determined the serotype of 977 S. pneumoniae isolates causing invasive disease between 2000-2002 and 2007-2008, assessed for the presence of the recently described serotype 6C and determined the susceptibility of isolates during 2007-2008 to penicillin and cefotaxime. Serotype 14 was the most common serotype during both periods and 7·7% of isolates previously typed as serotype 6A were serotype 6C. During 2000-2002 and 2007-2008, PCV7 could potentially have prevented 85% and 74% of invasive pneumococcal disease in the target population (i.e. children aged <2 years), respectively. The level of penicillin non-susceptibility was 17% in 2007-2008. Ongoing surveillance of serotypes is required to determine the impact of PCV7 in the Irish population and to assess the potential of new vaccines with expanded valency.

  17. Serotype and mating type characterization of Cryptococcus neoformans by multiplex PCR.

    Carvalho, Vívian Gonçalves; Terceti, Mateus Souza; Dias, Amanda Latercia Tranches; Paula, Claudete Rodrigues; Lyon, Juliana Pereira; de Siqueira, Antônio Martins; Franco, Marília Caixeta


    Cryptococcus neoformans is an encapsulated yeast, etiological agent of cryptococcosis. The species is commonly associated with pigeon droppings and plant materials. The aim of the present work was to verify the presence of the yeast in pigeon droppings, and to identify the isolates obtained in serotypes and mating types (MAT). Ten samples of pigeon droppings were collected in the rural area of the city of Alfenas, Brazil. Samples were inoculated in agar Niger medium for fungal isolation and 22 isolates with characteristics of C. neoformans were obtained. The serotypes and MAT were determined by multiplex PCR using specific primers. Serotypes were also determined by using the Kit Crypto Check. Among the 22 samples evaluated, eight were identified as C. neoformans by classic identification tests. These samples were characterized as serotype A by the Kit Crypto check and as serotype A MAT alpha by the multiplex PCR. The present study reinforces the evidence that pigeon droppings are a reservoir for C. neoformans and confirms the prevalence of C. neoformans var. grubii (A alpha) among environmental isolates. It also demonstrates that multiplex PCR is an acceptable alternative for serotype analysis because it reduces the costs for each reaction and analyses serotype and MAT simultaneously. PMID:17823747

  18. Serotypes and antimicrobial resistance of meningeal isolates of Streptococcus pneumonia. Cuba, 2007-2012

    Gilda Toraño-Peraza


    Full Text Available An observational study was conducted to know the serotypes and antimicrobial susceptibility of isolates of Streptococcus pneumoniae responsible for meningitis in Cuba, where there is no vaccine yet to prevent invasive pneumococcal disease. The study included the total number of isolates submitted to the "Pedro Kourí" Institute between 2007 and 2012 (N=237. Serotypes identification was performed using capsular swelling test and antimicrobial susceptibility was studied by determining the minimum inhibitory concentration using the broth microdilution method. Predominant serotypes were 6A, 6B, 14, 19F and 23F and other non-vaccinal 18 serogroups/serotypes were identified in 29.1% of the isolates. A tendency to an increased resistance to penicillin (44.3 % was observed; the most common resistance patterns were: penicillin-trimethoprim/sulfamethoxazole and penicillin-erythromycin (21.1% and 10.5%, respectively. The largest number of isolates resistant to penicillin was in serotypes 6B, 14, 19F and 23F and the possibility of resistant non-vaccine serotypes emergence should be considered. The results show that 70.4 % of the isolates studied corresponds to the serotypes included in 13-valent conjugated pneumococcal vaccine, but with 10-valent it would achieve a lower vaccination potential coverage (56.1%. This information must be considered when evaluating the decision to use in Cuba any commercially available vaccine or the proposal of another strategy of vaccination from autochthonous vaccine candidates.

  19. Molecular epidemiology of serotype 19A Streptococcus pneumoniae isolated from children in Beijing, 1997-2006

    XUE Lian; YAO Kai-hu; YU Sang-jie; LIU Zun-jie; QIAN Jing; SHEN Xu-zhuang; YANG Yong-hong


    Background Despite the prevalence of Streptococcus pneumoniae serotype 19A, the molecular characteristics of this serotype are yet to be fully elucidated. The aim of this study was therefore to determine the homology of the serotype 19A in China.Methods Pulsed-field gel electrophoresis and multilocus sequence typing were done to these forty-nine serotype 19A isolates to investigate the relationship between the strains prevalent in Beijing and other regions. Results From 1997 to 2006, the percentage of serotype 19A isolates increased. The susceptibility rate to penicillin and amoxicillin decreased and the resistance rate to cefuroxime increased. ST320 was the most prevalent ST, followed by ST3546. There were six new STs identified in our study. The serotype 19A strains were classified into six different pulsed-field gel electrophoresis (PFGE) patterns. ST320, which was associated with two different PFGE patterns (A and D), accounted for 32 isolates, and ST3546, which was associated with two PFGE patterns (B and E), accounted for eightConclusions From 2003 onwards, ST320 was the most common ST and the rate of resistance to cefuroxime increased significantly. Further long-term surveys of Streptococcus pneumoniae serotype 19A are required to monitor ST prevalence and antimicrobial resistance in this important human pathogen.

  20. VP2-serotyped live-attenuated bluetongue virus without NS3/NS3a expression provides serotype-specific protection and enables DIVA.

    Feenstra, Femke; Maris-Veldhuis, Mieke; Daus, Franz J; Tacken, Mirriam G J; Moormann, Rob J M; van Gennip, René G P; van Rijn, Piet A


    Bluetongue virus (BTV) causes Bluetongue in ruminants and is transmitted by Culicoides biting midges. Vaccination is the most effective measure to control vector borne diseases; however, there are 26 known BTV serotypes showing little cross protection. The BTV serotype is mainly determined by genome segment 2 encoding the VP2 protein. Currently, inactivated and live-attenuated Bluetongue vaccines are available for a limited number of serotypes, but each of these have their specific disadvantages, including the inability to differentiate infected from vaccinated animals (DIVA). BTV non-structural proteins NS3 and NS3a are not essential for virus replication in vitro, but are important for cytopathogenic effect in mammalian cells and for virus release from insect cells in vitro. Recently, we have shown that virulent BTV8 without NS3/NS3a is non-virulent and viremia in sheep is strongly reduced, whereas local in vivo replication leads to seroconversion. Live-attenuated BTV6 without NS3/NS3a expression protected sheep against BTV challenge. Altogether, NS3/NS3a knockout BTV6 is a promising vaccine candidate and has been named Disabled Infectious Single Animal (DISA) vaccine. Here, we show serotype-specific protection in sheep by DISA vaccine in which only genome segment 2 of serotype 8 was exchanged. Similarly, DISA vaccines against other serotypes could be developed, by exchange of only segment 2, and could therefore safely be combined in multi-serotype cocktail vaccines with respect to reassortment between vaccine viruses. Additionally, NS3 antibody responses are raised after natural BTV infection and NS3-based ELISAs are therefore appropriate tools for DIVA testing accompanying the DISA vaccine. To enable DIVA, we developed an experimental NS3 ELISA. Indeed, vaccinated sheep remained negative for NS3 antibodies, whereas seroconversion for NS3 antibodies was associated with viremia after heterologous BTV challenge. PMID:25454873

  1. Serotype b of Aggregatibacter actinomycetemcomitans increases osteoclast and memory T-lymphocyte activation.

    Melgar-Rodríguez, S; Díaz-Zúñiga, J; Alvarez, C; Rojas, L; Monasterio, G; Carvajal, P; Escobar, A; Sanz, M; Vernal, R


    During periodontitis, alveolar bone resorption is associated with activation of T helper type 17 (Th17) lymphocytes and receptor activator of nuclear factor-κB ligand (RANKL) -induced osteoclasts. We previously reported that serotype b of Aggregatibacter actinomycetemcomitans has a higher capacity to trigger Th17-type differentiation and function in activated T lymphocytes and its lipopolysaccharide is a more potent immunogen compared with the other serotypes. This study aimed to investigate whether serotype b of A. actinomycetemcomitans induces higher Th17-associated RANKL production, RANKL-induced osteoclast activation, and antigen-specific memory T lymphocyte proliferation. On naive CD4(+) T lymphocytes stimulated with autologous dendritic cells primed with different A. actinomycetemcomitans serotypes, RANKL production, T-bet, GATA-3, RORC2 and Foxp3 expression, RORC2/RANKL intracellular double-expression, TRAP(+) osteoclast activation, and bone resorption were quantified. The frequency of proliferating memory T lymphocytes in response to A. actinomycetemcomitans serotypes was determined in periodontitis and healthy subjects. Naive CD4(+) T lymphocytes stimulated by serotype b-primed dendritic cells elicited higher levels of RANKL, RORC2, TRAP(+) osteoclasts, and bone resorption than the same cells stimulated with the other serotypes. RANKL positively correlated and co-expressed with RORC2. Memory T lymphocytes responding to serotype b were more frequently detected in periodontitis patients than healthy subjects. These results indicate that serotype b of A. actinomycetemcomitans is associated with higher production of RANKL and these increased levels are associated with Th17 lymphocyte induction, osteoclast activation, and bone resorption. PMID:26172400

  2. Serotype variation among infectious bronchitis viral isolates taken from several areas of Java

    Risa Indriani


    Full Text Available Infectious bronchitis (IB is an acute highly contagious viral respiratory disease of poultry caused by virus belongs to the family of Coronaviridae. The virus consist of many serotypes with low level of cross-protectivity among serotypes. Field data showed that the outbreaks of IB were frequently reported in chicken flocks, although vaccinations against the disease have been practiced. Hence, the study on serotype relationship among isolates of the viruses is essentially required. The aim of this study was to isolate and characterize IB viruses from chicken flocks in some areas of Java. Isolation of the virus was carried out in nine-day old embrionated chicken eggs and identified by means of agar gel precipitation (AGP tests against standard antisera to IB virus. The serotypes of the IB viral isolates were determined by cross-neutralization tests in nine day old embryonated chicken eggs using r value derived from homologous and heterologous serum titres as criteria. This study obtained 12 IB viral isolates which were identified on the basis of the ability to cause lesions in chicken embryos and positive to agar gel presipitation test against standard positive antiserum to the virus. Based on the cross-neutralization tests in embryonated chicken eggs, isolate I.9 was formed to have relationship closed to Mass-41 serotype, while I.2, I. 3, and I.7 isolates were closely to the serotype of Con-46. Virus isolates (I.5, I.14, I.24, and I.25 were decided to have no serotype relationships to either Mass-41 or Con-46 serotype. Since the I.5, I.14, I.24 and I.25 isolates were not neutralized by antisera against the previous identified local infectious bronchitis viral isolates, and that were considered to be distinct serotype to the previously identified local IB viral isolates.

  3. [Detection and Serotyping of Streptococcus pneumoniae Carried in Healthy Adults with a Modified PCR Method].

    Ishihara, Yuka; Okamoto, Akira; Ohta, Michio


    Detection of Streptococcus pneumoniae colonized in the pharynx of healthy carriers currently relies on conventional culture methods of direct plating with pharyngeal swab specimens. The accurate measurement of the carriage of pneumococci, however, has not been necessarily achieved with these methods due to low density colonization and contamination of numerous oral streptococci that express α-hemolysis. A PCR-based detection method of pneumococci-specific for lytA as well as PCR serotyping of S. pneumoniae was recently developed and their effectiveness was confirmed. We modified the reaction conditions of these methods to improve the detection rate and applied them to the measurement of S. pneumoniae carried in healthy adults. Pharyngeal swab specimens obtained from 110 healthy volunteers over 40 and living in Nagoya were enriched for 5 hours with broth medium supplemented with rabbit serum and the template DNA for PCR was extracted from the mixed enriched culture. Of 110 specimens 36 (32.7%) were lytA-positive, the rate of which was much higher than the results of previous culture-based studies. The DNA template preparations were then used for PCR-based serotyping with primers specific for each of the types included in pneumococcal 23 valent vaccine (PPV23). We found that 28 out of 36 lytA-positive carriers were identified as being positive for the serotypes belonging to PPV23, although serotypes 6A and 6B were indistinguishable with the PCR method. The most frequent serotype was serotype 14, and serotypes 4, 18C, and 6A/B were also frequently identified. Five lytA-positive carriers were previously vaccinated with PPV23, and among them, 4 were positive for serotypes contained in PPV23. We recommend PCR-based identification and serotyping of S. pneumoniae in broth enrichment culture of pharyngeal swab specimens as a reliable method for the surveillance of healthy carriers with low density colonization. PMID:26552129

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

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


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

  5. Chemical Analysis of Major Outer Membrane Proteins of Neisseria meningitidis: Comparison of Serotypes 2 and 11

    Tsai, Chao-Ming; Frasch, Carl E.


    Most of the 15 protein serotypes found in group B Neisseria meningitidis have distinct major outer membrane protein patterns when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) by the Weber-Osborn system. Both serotypes 2 and 11 contain major outer membrane proteins with apparent molecular weights of 41,000 and 28,000 (41K and 28K). The 41K and 28K proteins were purified from the prototype strains of these two serotypes (M986 type 2 and M136 type 11) by prepa...

  6. Clonal relationship of recent invasive Haemophilus influenzae serotype f isolates from Denmark and the United States

    Bruun, B; Gahrn-Hansen, B; Westh, H;


    Surveillance performed after the introduction of general Haemophilus influenzae serotype b (Hib) vaccination in Denmark identified 13 cases of invasive bacteraemic H. influenzae serotype f (Hif) disease in adults over a period of 7 years. Bacteraemic respiratory tract infections accounted for 61......A gene sequences. Multilocus enzyme electrophoresis typing revealed that recent Danish and American isolates belonged to a single Hif clone, which may be undergoing expansion. The need for accurate serotyping of H. influenzae to enable reliable monitoring for Hib replacement by other capsular types is...

  7. Serotypes of Streptococcus agalactiae cultured from dairy milk samples in Québec

    Daignault, Danielle; Guévremont, Evelyne; Guillemette, Jean-Marc; Messier, Serge; Gottschalk, Marcelo; Higgins, Robert


    Streptococcus agalactiae remains an important pathogen of dairy herds in Québec, but data about antigenic characteristics of this microorganism are sparse. This study was conducted to determine the variety of S. agalactiae serotypes in dairy herds in Québec. Two hundred and ninety-five isolates cultured from the milk of individual cows from 7 regions of Québec were serotyped. Sixty-two percent of the isolates were untypeable. Among the 38% of typeable isolates, serotype III was found most fre...

  8. Dicty_cDB: Contig-U15214-1 [Dicty_cDB

    Full Text Available e) Actinobacillus pleuropneumoniae... 107 4e-22 CP000927_3202( CP000927 |pid:none) Cau...lobacter sp. K31, complete g... 107 4e-22 CP000569_1501( CP000569 |pid:none) Actinobacillus pleuropneumoniae...000687_1497( CP000687 |pid:none) Actinobacillus pleuropneumoniae... 107 6e-22 (Q9...P001098_708( CP001098 |pid:none) Halothermothrix orenii H 168, co... 104 4e-21 (Q8RQM6) RecName: Full=Diaminopim... 83 9e-15 AE017283_1241( AE017283 |pid:none) Propionibacterium acnes KPA1712... 83 9e-15 (Q9X5M1) RecName: Full=Diaminopim

  9. Streptococcus pneumoniae Serotype Distribution and Pneumococcal Conjugate Vaccine Serotype Coverage among Pediatric Patients in East and Southeast Asia, 2000–2014: a Pooled Data Analysis

    Stanley S. Tai


    Full Text Available Pneumococcal infection is one of the leading causes of death worldwide, especially in children of developing and underdeveloped countries. Capsular polysaccharide-based vaccines are available for the prevention of this disease. A 7-valent pneumococcal conjugate vaccine (PCV7 was licensed in 2000 for use in children less than two years of age. Subsequently, to broaden the protection, 10-valent (PCV10 and 13-valent (PCV13 vaccines were licensed in 2009 and 2010, respectively. All of these conjugate vaccines elicit an immune response that only provides protection against the infection of S. pneumoniae serotypes included in the formulation. Profiles of S. pneumoniae serotype distribution and serotype coverage for both PCV7 and PCV13 have been reported in some Asian countries/territories. But the published results cannot provide conclusive information due to the difference in studied population and geographic areas. The goals of this review are to obtain an accurate estimate of serotype coverage for PCV7, PCV10, and PCV13 and examine the change in the S. pneumoniae serotype distribution after PCV7 use among pediatric patients in East and Southeast Asia through the analysis of pooled data that were published in the English literature between 2000 and 2014.

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

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


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

  11. Molecular serotyping of rough Salmonella strains isolated from Danish pork production

    Löfström, Charlotta; Boel, Jeppe; Sisic, E.;


    Salmonella is an important zoonotic pathogen and meat, including pork, is one of the main sources of salmonellosis. Surveillance of Salmonella in the meat production chain is therefore essential to increase food safety. To enable e.g. source attribution and to reveal the source of outbreaks methods...... incomplete LPS structures, referred to as rough strains, or do not express H antigens, therefore serotyping by agglutination cannot be performed on these isolates. This results in data gaps when subtyping data is needed. To overcome this obstacle, serotypes can be determined on DNA level, i.e. by determining...... the same time period from similar sources. A number of rough Salmonella strains (n = 211) isolated from pig carcasses during 2005-2012 were analyzed with molecular serotyping employing Luminex technology. Results show that molecular serotyping enabled serovar identification in 168 of the strains (80...

  12. Genome sequences of mannheimia haemolytica serotype A2: ovine and bovine isolates

    This report describes the genome sequences of Mannheimia haemolytica, serotype A2 isolated from pneumonic lungs of two different ruminant species, one from Ovis aries, designated as Ovine (O) and the other from Bos taurus, designated as Bovine (B)....

  13. The Widespread Multidrug-Resistant Serotype O12 Pseudomonas aeruginosa Clone Emerged through Concomitant Horizontal Transfer of Serotype Antigen and Antibiotic Resistance Gene Clusters

    Thrane, Sandra Wingaard; Taylor, Véronique L.; Freschi, Luca;


    conclusion, serotype switching in combination with acquisition of an antibiotic resistance determinant most likely contributed to the dissemination of the O12 serotype in clinical settings. Infection rates in hospital settings by multidrug-resistant (MDR) Pseudomonas aeruginosa clones have increased during...... switching was the result of horizontal transfer and genetic recombination of lipopolysaccharide (LPS) biosynthesis genes originating from an MDR taxonomic outlier P. aeruginosa strain. Moreover, the recombination event also resulted in acquisition of antibiotic resistance genes. These results impact on our...... clinical settings and outbreaks. These serotype O12 isolates exhibit high levels of resistance to various classes of antibiotics. Here, we explore how the P. aeruginosa OSA biosynthesis gene clusters evolve in the population by investigating the association between the phylogenetic relationships among 83 P...

  14. Detection and transmission of extracellular fac-tor producing Streptococcus suis serotype 2 strains in pigs

    Swildens, B.


    DETECTION AND TRANSMISSION OF EXTRACELLULAR FACTOR PRODUCING STREPTOCOCCUS SUIS SEROTYPE 2 STRAINS IN PIGS INTRODUCTION Streptococcus suis (S.suis) has been implicated in the etiology of many diseases among which meningitis in pigs. The virulent extracellular factor-positive strains of S.suis serotype 2 (S.suis2EF+) have frequently been isolated from diseased. Sows carrying S.suis strains in their tonsils are considered the most important source of infection for their susceptible offspring. D...

  15. Correlation of Serotype-Specific Dengue Virus Infection with Clinical Manifestations

    Halsey, Eric S; Marks, Morgan A.; Gotuzzo, Eduardo; Fiestas, Victor; Suarez, Luis; Vargas, Jorge; Aguayo, Nicolas; Madrid, Cesar; Vimos, Carlos; Kochel, Tadeusz J.; Laguna-Torres, V. Alberto


    Background Disease caused by the dengue virus (DENV) is a significant cause of morbidity throughout the world. Although prior research has focused on the association of specific DENV serotypes (DENV-1, DENV-2, DENV-3, and DENV-4) with the development of severe outcomes such as dengue hemorrhagic fever and dengue shock syndrome, relatively little work has correlated other clinical manifestations with a particular DENV serotype. The goal of this study was to estimate and compare the prevalence ...

  16. Coexistence of two dengue virus serotypes and forecasting for Madeira island

    Rocha, Filipa Portugal; Rodrigues, Helena Sofia; Monteiro, M. Teresa T.; Torres, Delfim F. M.


    The first outbreak of dengue occurred in Madeira Island on 2012, featuring one virus serotype. Aedes aegypti was the vector of the disease and it is unlikely that it will be eliminated from the island. Therefore, a new outbreak of dengue fever can occur and, if it happens, risk to the population increases if two serotypes coexist. In this paper, mathematical modeling and numerical simulations are carried out to forecast what may happen in Madeira Island in such scenario.

  17. Vibrio parahaemolyticus Strains of Pandemic Serotypes Identified from Clinical and Environmental Samples from Jiangsu, China

    Jingjiao eLi; Feng eXue; Zhenquan eYang; Xiaoping eZhang; Dexin eZeng; Guoxiang eChao; Yuan eJiang; Baoguang eLi


    Vibrio parahaemolyticus has emerged as a major foodborne pathogen in China, Japan, Thailand and other Asian countries. In this study, 72 strains of V. parahaemolyticus were isolated from clinical and environmental samples between 2006 and 2014 in Jiangsu, China. The serotypes and six virulence genes including thermostable direct hemolysin (TDR) and TDR-related hemolysin (TRH) genes were assessed among the isolates. Twenty five serotypes were identified and O3:K6 was one of the dominant seroty...

  18. Association of Three Plasmid-Encoded spv Genes Among Different Salmonella Serotypes Isolated from Different Origins

    Derakhshandeh, Abdollah; Firouzi, Roya; Khoshbakht, Rahem


    The virulence plasmid associated Salmonella plasmid virulence (spv) locus is strongly concomitant with strains that cause non typhoid bacteremia. The spv region contains three genes required for the virulence, the positive transcriptional regulator spvR and two structural genes spvB and spvC. The purpose of this study was to investigate the presence of these three genes among salmonella serotypes isolated from different sources. A collection of 60 salmonella serotypes from different sources w...

  19. Characterization of serological cross-reactivity between polysaccharide antigens of Streptococcus mutans serotypes c and d.

    Grossi, S.; Prakobphol, A; Linzer, R; Campbell, L K; Knox, K W


    Immunological assays with antisera prepared against purified Streptococcus mutans serotype c polysaccharide demonstrated that a cross-reacting determinant on c polysaccharide reacted with the wall-associated rhamnose-glucose polysaccharide from S. mutans serotype d. Studies with 60 antisera prepared against chemostat cultures of S. mutans Ingbritt (c) demonstrated that the rhamnose-glucose polysaccharide cross-reactive determinant was consistently expressed on c antigen under a variety of gro...

  20. Vibrio parahaemolyticus Strains of Pandemic Serotypes Identified from Clinical and Environmental Samples from Jiangsu, China

    Jingjiao eLi


    Full Text Available Vibrio parahaemolyticus has emerged as a major foodborne pathogen in China, Japan, Thailand and other Asian countries. In this study, 72 strains of V. parahaemolyticus were isolated from clinical and environmental samples between 2006 and 2014 in Jiangsu, China. The serotypes and six virulence genes including thermostable direct hemolysin (TDR and TDR-related hemolysin (TRH genes were assessed among the isolates. Twenty five serotypes were identified and O3:K6 was one of the dominant serotypes. The genetic diversity was assessed by multilocus sequence typing (MLST analysis, and 48 sequence types (STs were found, suggesting this V. parahaemolyticus group is widely dispersed and undergoing rapid evolution. A total of 25 strains of pandemic serotypes such as O3:K6, O5:K17 and O1:KUT were identified. It is worth noting that the pandemic serotypes were not exclusively identified from clinical samples, rather, nine strains were also isolated from environmental samples; and some of these strains harbored several virulence genes, which may render those strains pathogenicity potential. Therefore, the emergence of these environmental pandemic V. parahaemolyticus strains may poses a new threat to the public health in China. Furthermore, six novel serotypes and 34 novel STs were identified among the 72 isolates, indicating that V. parahaemolyticus were widely distributed and fast evolving in the environment in Jiangsu, China. The findings of this study provide new insight into the phylogenic relationship between V. parahaemolyticus strains of pandemic serotypes from clinical and environmental sources and enhance the MLST database; and our proposed possible O- and K- antigen evolving paths of V. parahaemolyticus may help understand how the serotypes of this dispersed bacterial population evolve.

  1. Simple, Rapid Latex Agglutination Test for Serotyping of Pneumococci (Pneumotest-Latex)

    Slotved, H.-C.; Kaltoft, M.; Skovsted, I. C.; Kerrn, M. B.; Espersen, F


    The “gold standard” for epidemiological typing of Streptococcus pneumoniae (pneumococcus) is the capsular reaction test (Neufeld test) with antisera against the 90 pneumococcal polysaccharide capsules, i.e., serotyping. The method is labor intensive and requires a certain level of experience to be performed satisfactory, and thus it has been restricted for use in specialized reference or research laboratories. Surveillance of the serotype distribution of pneumococci that cause infections is i...

  2. Diagnosis of Invasive Pneumococcal Infection by Serotype-Specific Urinary Antigen Detection

    Leeming, John P.; Cartwright, Keith; Morris, Rhonwen; Martin, Siobhan A.; Smith, Michael D.


    Widespread use of conjugate pneumococcal polysaccharide-protein vaccines may alter the spectrum of pneumococci producing invasive disease. Novel sensitive diagnostic methods would be valuable for monitoring the epidemiology of pneumococcal disease within populations and vaccine recipients. Ideally, these methods should allow determination of the serotype of the infecting clone. Serotype-specific enzyme-linked immunosorbent assays (ELISA) for 13 capsular polysaccharides (types 1, 3, 4, 5, 6A, ...

  3. Impact of Preceding Flu-Like Illness on the Serotype Distribution of Pneumococcal Pneumonia

    Song, Joon Young; Nahm, Moon H.; Cheong, Hee Jin; Kim, Woo Joo


    Background Even though the pathogenicity and invasiveness of pneumococcus largely depend on capsular types, the impact of serotypes on post-viral pneumococcal pneumonia is unknown. Methods and Findings This study was performed to evaluate the impact of capsular serotypes on the development of pneumococcal pneumonia after preceding respiratory viral infections. Patients with a diagnosis of pneumococcal pneumonia were identified. Pneumonia patients were divided into two groups (post-viral pneumococcal pneumonia versus primary pneumococcal pneumonia), and then their pneumococcal serotypes were compared. Nine hundred and nineteen patients with pneumococcal pneumonia were identified during the study period, including 327 (35.6%) cases with post-viral pneumococcal pneumonia and 592 (64.4%) cases with primary pneumococcal pneumonia. Overall, serotypes 3 and 19A were the most prevalent, followed by serotypes 19F, 6A, and 11A/11E. Although relatively uncommon (33 cases, 3.6%), infrequently colonizing invasive serotypes (4, 5, 7F/7A, 8, 9V/9A, 12F, and 18C) were significantly associated with preceding respiratory viral infections (69.7%, P<0.01). Multivariate analysis revealed several statistically significant risk factors for post-viral pneumococcal pneumonia: immunodeficiency (OR 1.66; 95% CI, 1.10–2.53), chronic lung diseases (OR 1.43; 95% CI, 1.09–1.93) and ICI serotypes (OR 4.66; 95% CI, 2.07–10.47). Conclusions Infrequently colonizing invasive serotypes would be more likely to cause pneumococcal pneumonia after preceding respiratory viral illness, particularly in patients with immunodeficiency or chronic lung diseases. PMID:24691515

  4. Serotype Specific Invasive Capacity and Persistent Reduction in Invasive Pneumococcal Disease

    Yildirim, Inci; William P Hanage; Lipsitch, Marc; Shea, Kimberly M.; STEVENSON, ABBIE; Finkelstein, Jonathan; Huang, Susan S.; Grace M Lee; Kleinman, Ken; Pelton, SI


    Defining the propensity of Streptoccocus pneumoniae (SP) serotypes to invade sterile body sites following nasopharyngeal (NP) acquisition has the potential to inform about how much invasive pneumococcal disease (IPD) may occur in a typical population with a given distribution of carriage serotypes. Data from enhanced surveillance for IPD in Massachusetts children ≤7 years in 2003/04, 2006/07 and 2008/09 seasons and surveillance of SP NP carriage during the corresponding respiratory seasons in...

  5. Associations of Streptococcus suis serotype 2 ribotype profiles with clinical disease and antimicrobial resistance

    Rasmussen, S. R.; Aarestrup, Frank Møller; Jensen, N. E.;


    A total of 122 Streptococcus suis serotype 2 strains were characterized thoroughly by comparing clinical and pathological observations, ribotype profiles, and antimicrobial resistance. Twenty-one different ribotype profiles were found and compared by cluster analysis, resulting in the identificat......A total of 122 Streptococcus suis serotype 2 strains were characterized thoroughly by comparing clinical and pathological observations, ribotype profiles, and antimicrobial resistance. Twenty-one different ribotype profiles were found and compared by cluster analysis, resulting in the...

  6. Changing pattern of dengue virus serotypes circulating during 2008-2012 and reappearance of dengue serotype 3 may cause outbreak in Kolkata, India.

    Saha, Kallol; Ghosh, Monika; Firdaus, Rushna; Biswas, Aritra; Seth, Bikash; Bhattacharya, Debojyoti; Mukherjee, Kheya; Sadhukhan, Provash Chandra


    Dengue virus infection is a major cause of morbidity within the endemic tropical and subtropical regions of the world. Dengue virus has four distinct serotypes with specific clinical manifestations. In this study, we observed the changing pattern of dengue serotypes, age-wise dengue infection and useful sero-detection methods needed in a dengue endemic region. We identified dengue serotypes during a period of 5 years among patients with dengue symptoms visiting one of the largest tertiary care infectious disease hospitals of eastern India in Kolkata. A total of 433 dengue RNA positive samples were isolated from 712 acute dengue suspected cases. Age wise distribution highlighted the susceptible age group being >21 years (24.02%) followed by 11-15 years (21.71%) and 5-10 years (21.02%) of the total infected population. Higher numbers of infected cases were found within females as they are involved in more indoor works. The period of study experienced two dengue outbreaks one in 2008 and another in 2012. For early dengue detection, NS1 was found to be more confirmatory than IgM ELISA regarding sensitivity and specificity. DENV-1, 2, and 4 serotypes were the common circulating strains from 2008 until 2010, after which DENV-3 serotype infections rise and led to a massive dengue outbreak in Kolkata with increased numbers of DHF and DSS cases in 2012. The finding within our study emphasizes the public health importance of such prospective surveillance programs with respect to the changing dengue viral etiology and serotypes. J. Med. Virol. 88:1697-1702, 2016. © 2016 Wiley Periodicals, Inc. PMID:26991505

  7. Simultaneous Rapid Detection and Serotyping of Cronobacter sakazakii Serotypes O1, O2, and O3 by Using Specific Monoclonal Antibodies.

    Scharinger, Eva J; Dietrich, Richard; Kleinsteuber, Ina; Märtlbauer, Erwin; Schauer, Kristina


    Cronobacter sakazakiiis a foodborne pathogen associated with rare but often lethal infections in neonates. Powdered infant formula (PIF) represents the most frequent source of infection. Out of the identified serotypes (O1 to O7), O1, O2, and O3 are often isolated from clinical and PIF samples. Serotype-specific monoclonal antibodies (MAbs) suitable for application in enzyme immunoassays (EIAs) for the rapid detection ofC. sakazakiihave not yet been developed. In this study, we created specific MAbs with the ability to bind toC. sakazakiiof serotypes O1, O2, and O3. Characterization by indirect EIAs, immunofluorescence, motility assays, and immunoblotting identified lipopolysaccharide (LPS) and exopolysaccharide (EPS) as the antigenic determinants of the MAbs. The established sandwich EIAs were highly sensitive and were able to detect between 2 × 10(3)and 9 × 10(6)CFU/ml. Inclusivity tests confirmed that 93% of serotype O1 strains, 100% of O2 strains, and 87% of O3 strains were detected at low cell counts. No cross-reactivity with >100 strains ofCronobacterspp. and otherEnterobacteriaceaewas observed, except for that withC. sakazakiiserotype O3 andCronobactermuytjensiiserotype O1. Moreover, the sandwich EIAs detectedC. sakazakiiin PIF samples artificially contaminated with 1 to 10 bacterial cells per 10 g of sample after 15 h of preenrichment. The use of these serotype-specific MAbs not only allows the reliable detection ofC. sakazakiistrains but also enables simultaneous serotyping in a simple sandwich EIA method. PMID:26850303

  8. Qualitative and quantitative impacts assessment of contagious bovine pleuropneumonia in Fulani pastoral herds of North-central Nigeria: The associated socio-cultural factors.

    Alhaji, N B; Babalobi, O O


    Contagious bovine pleuropneumonia is one of the most important trans-boundary disease affecting Fulani cattle herds of Nigeria and whose control is urgently needed. A Participatory Epidemiology approach and cross-sectional study were concurrently conducted to investigate qualitative and quantitative impacts of CBPP, respectively and associated socio-cultural factors that influenced exposure of Fulani nomadic pastoral communities to its risk in Niger State, North-central Nigeria between January and December 2013. A total of nine pastoral communities were purposively selected for qualitative impact assessment using Participatory Rural Appraisal tools, while 765 cattle randomly sampled from 125 purposively selected nomadic herds were analyzed using c-ELISA. Data on socio-cultural characteristics were collected using structured questionnaires administered on nomadic herd owners of the 125 selected herds. Kendall's Coefficient of Concordance W statistics and OpenEpi 2.3 were used for statistical analyses. Pastoralists' dependent factors associated with their socio-cultural activities were tested using Chisquare tests and likelihood backward logistic regressions. The mean proportional piles (relative qualitative impact) of CBPP was 12.6%, and nomads agreement on this impact was strong (W=0.6855) and statistically significant (Psource that caused concentration of stocks in one point was fifty three times more likely (OR 53.08; 95% CI: 14.91, 189.00) to be satisfactory practice that influenced occurrence of the disease in herds. This study highlighted the critical gap that exists in terms of significant influence of socio-cultural factors on CBPP occurrence in pastoral herds in Nigeria. Thus, CBPP surveillance, control and prevention programs that take these factors into consideration will be beneficial to the livestock industry in Nigeria, and indeed Africa. PMID:27237398

  9. Chlamydia trachomatis serotype A infections in the Amazon region of Brazil: prevalence, entry and dissemination

    Marluísa de Oliveira Guimarães Ishak


    Full Text Available INTRODUCTION: Chlamydia infection is associated with debilitating human diseases including trachoma, pneumonia, coronary heart disease and urogenital diseases. Serotypes of C. trachomatis show a fair correlation with the group of diseases they cause, and their distribution follows a well-described geographic pattern. Serotype A, a trachoma-associated strain, is known for its limited dissemination in the Middle East and Northern Africa. However, knowledge on the spread of bacteria from the genus Chlamydia as well as the distribution of serotypes in Brazil is quite limited. METHODS: Blood samples of 1,710 individuals from ten human population groups in the Amazon region of Brazil were examined for antibodies to Chlamydia using indirect immunofluorescence and microimmunofluorescence assays. RESULTS: The prevalence of antibodies to Chlamydia ranged from 23.9% (Wayana-Apalai to 90.7% (Awa-Guaja with a mean prevalence of 50.2%. Seroreactivity was detected to C. pneumoniae and to all serotypes of C. trachomatis tested; furthermore, we report clear evidence of the as-yet-undescribed occurrence of serotype A of C. trachomatis. CONCLUSIONS: Specific seroreactivity not only accounts for the large extent of dissemination of C. trachomatis in the Amazon region of Brazil but also shows an expanded area of occurrence of serotype A outside the epidemiological settings previously described. Furthermore, these data suggest possible routes of Chlamydia introduction into the Amazon region from the massive human migration that occurred during the 1,700s.

  10. Serotype and serovar distribution of Neisseria gonorrhoeae isolated from high-risk populations in Bangladesh.

    Alam, M A; Chowdhury, M Z; Ahmed, F; Alam, A; Hossain, M A


    Neisseria gonorrhoeae, the causative agent of gonococcal infection, is known to frequently change their characteristics to evade host immune mechanism. Characterization of the clinical isolates of the organism can lead to identification of the circulating strains and often a sexual network in a community to help in designing the control strategy. Keeping in mind the above consideration, a total of 239 N. gonorrhoeae, isolated from high-risk populations, were characterized for serotypes and serovars by monoclonal antibodies against protein 1 of the organism. Majority of the serotypes were serotype B (142, 59.4%). Majority of the isolates showing resistance to at least one of the antibiotics tested were also serotype B (139, 59.2%), whereas, majority of the isolates showing resistance to any three of the antibiotics (multidrug resistant, MDR) (63%) was serotype A. A total of 41 different serovars were also identified and five of which (Arst, Bropt, Bopt, Arost, and Brop) included the highest percent (49.3%) of the isolates. Many serovars (23/41, 56.1%) were new emergent and included 58 (24.3%) of the isolates investigated. All of the new serovars were resistant to at least one of the antibiotics tested and the highest rate (40/102, 39.2%) was MDR. Serotyping and serovar determination was found contributory to understand the microepidemics of the N. gonorrhoeae isolates. Further studies including antibiogram and contact tracing can efficiently help in control of the disease. PMID:23540188

  11. Rapid and Easy In Silico Serotyping of Escherichia coli Isolates by Use of Whole-Genome Sequencing Data

    Joensen, Katrine Grimstrup; Tetzschner, Anna M. M.; Iguchi, Atsushi;


    typing and surveillance. The aim of this study was to establish a valid and publicly available tool for WGS-based in silico serotyping of E. coli applicable for routine typing and surveillance. A FASTA database of specific O-antigen processing system genes for O typing and flagellin genes for H typing...... was created as a component of the publicly available Web tools hosted by the Center for Genomic Epidemiology (CGE) ( All E. coli isolates available with WGS data and conventional serotype information were subjected to WGS-based serotyping employing this specific Serotype......Finder CGE tool. SerotypeFinder was evaluated on 682 E. coli genomes, 108 of which were sequenced for this study, where both the whole genome and the serotype were available. In total, 601 and 509 isolates were included for O and H typing, respectively. The O-antigen genes wzx, wzy, wzm, and wzt and the...

  12. Spatial Trend of Foot and Mouth Disease Virus (FMDV) Serotypes in Cattle and Buffaloes, Pakistan

    Muhammad Abubakar; Muhammad Javed Arshed; Qurban Ali; Manzoor Hussain


    The present study describes the frequency of Foot and Mouth Disease (FMD) virus serotypes (O,A and Asia-1) in major regions (all provinces) of Pakistan using Indirect Sandwich ELISA.Also,spatial distribution of various FMD serotypes and their comparison is discussed.A total of 590 samples (Epithelial tissue) have been analyzed during a period of five years (2005-2009).Out of 590 samples,180 were found positive,giving an overall confirmation of FMDV about 33.2 %.Of the prevalent serotypes,FMDV ‘O’ serotype caused most outbreaks (20.7 %),followed by serotype A (6.6 %) and serotype Asia-1 (4.6 %) while there was no positive case of type ‘C’.The study clearly showed that the disease was more frequent in the agro-climatic zones than in hilly areas.Based on the data of 590 samples (>50 outbreaks),the overall prevalence of FMDV in cattle and buffaloes in Pakistan was 33.2 %,while in cattle alone,it was 37.1%,higher than in buffalo (28.7 %).There were eight cases of mixed serotypes infection,indicating the presence of endemic state of disease.Another significant feature was the change over time.In phase-I (2005-2007),there was an overall prevalence of 29.4 %,while the occurrence of the serotype O,A and Asia-1 was 20.4 %,2.9 % and 4.7 %,respectively.During phase-II (2008-2009),the overall prevalence was 59.21%,while those of serotype O,A and Asia-1 were 22.4 %,31.6 % and 4.0 %,respectively.This clearly indicated a shift from serotype O to A,which may help to explain the occurrence of more severe outbreaks,despite vaccination.

  13. Identification of a natural human serotype 3 parainfluenza virus

    Wang Xiao-Jing


    Full Text Available Abstract Parainfluenza virus is an important pathogen threatening the health of animals and human, which brings human many kinds of disease, especially lower respiratory tract infection involving infants and young children. In order to control the virus, it is necessary to fully understand the molecular basis resulting in the genetic diversity of the virus. Homologous recombination is one of mechanisms for the rapid change of genetic diversity. However, as a negative-strand virus, it is unknown whether the recombination can naturally take place in human PIV. In this study, we isolated and identified a mosaic serotype 3 human PIV (HPIV3 from in China, and also provided several putative PIV mosaics from previous reports to reveal that the recombination can naturally occur in the virus. In addition, two swine PIV3 isolates transferred from cattle to pigs were found to have mosaic genomes. These results suggest that homologous recombination can promote the genetic diversity and potentially bring some novel biologic characteristics of HPIV.

  14. Rapid typing of foot-and-mouth disease serotype Asia 1 by reverse transcription loop-mediated isothermal amplification

    Chen Hao-tai; Zhang Jie; Liu Yong-sheng; Liu Xiang-tao


    Abstract A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was rapidly used to detect serotype Asia 1 of foot-and-mouth disease virus (FMDV) within 45 min at 61°C. All FMDV serotype Asia 1 reference strains were positive by RT-LAMP, while other viruses such as FMDV serotypes O, C, A and classical swine fever virus, swine vesicular disease virus, porcine reproductive and respiratory syndrome virus and Japanese encephalitis virus remained negative. Furthermore, FMDV...

  15. Identification of a high-virulence clone of serotype III Streptococcus agalactiae by growth characteristics at 40 degrees C.

    Mattingly, S J; Maurer, J J; Eskew, E K; Cox, F


    A high-virulence clone of serotype III Streptococcus agalactiae causing invasive neonatal disease was previously identified by multilocus enzyme electrophoresis. A simple procedure involving growth at 40 degrees C distinguished all isolates classified in this high-virulence clone from other serotype III isolates, which are more frequently associated with asymptomatically colonized infants, as well as the other serotypes of group B streptococci. The high-virulence clone failed to grow at 40 de...

  16. Dengue virus-specific human T cell clones. Serotype crossreactive proliferation, interferon gamma production, and cytotoxic activity


    The severe complications of dengue virus infections, hemorrhagic manifestation and shock, are much more commonly observed during secondary infections caused by a different serotype of dengue virus than that which caused the primary infections. It has been speculated, therefore, that dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) are caused by serotype crossreactive immunopathological mechanisms. We analyzed clones of dengue serotype crossreactive T lymphocytes derived from the...

  17. Concurrent infections by all four dengue virus serotypes during an outbreak of dengue in 2006 in Delhi, India

    Guleria Randeep; Dar Lalit; Diddi Kavita; Pandey Anubhav; Chahar Harendra S; Bharaj Preeti; Kabra Sushil K; Broor Shobha


    Abstract Background Co-circulation of multiple dengue virus serotypes has been reported from many parts of the world including India, however concurrent infection with more than one serotype of dengue viruses in the same individual is rarely documented. An outbreak of dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) occurred in and around Delhi in 2006. This is the first report from India with high percentage of concurrent infections with different dengue virus serotypes circulating d...

  18. Development and Characterization of a Reverse Genetic System for Studying Dengue Virus Serotype 3 Strain Variation and Neutralization

    Messer, William B.; Boyd Yount; Kari E Hacker; Donaldson, Eric F.; Huynh, Jeremy P.; de Silva, Aravinda M.; Baric, Ralph S.


    Dengue viruses (DENV) are enveloped single-stranded positive-sense RNA viruses transmitted by Aedes spp. mosquitoes. There are four genetically distinct serotypes designated DENV-1 through DENV-4, each further subdivided into distinct genotypes. The dengue scientific community has long contended that infection with one serotype confers lifelong protection against subsequent infection with the same serotype, irrespective of virus genotype. However this hypothesis is under increased scrutiny an...

  19. Role of RpoS in Fine-Tuning the Synthesis of Vi Capsular Polysaccharide in Salmonella enterica Serotype Typhi▿

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


    Regulation of the synthesis of Vi polysaccharide, a major virulence determinant in Salmonella enterica serotype Typhi, is under the control of two regulatory systems, ompR-envZ and rscB-rscC, which respond to changes in osmolarity. Some serotype Typhi strains exhibit overexpression of Vi polysaccharide, which masks clinical detection of lipopolysaccharide O antigen. This variation in Vi polysaccharide and O antigen display (VW variation) has been observed since the initial studies of serotype...

  20. Distribution and content of class 1 integrons in different Vibrio cholerae O-serotype strains isolated in Thailand

    Dalsgaard, Anders; Forslund, Anita; Serichantalergs, Oralak;


    In this study, 176 clinical and environmental Vibrio cholerae strains of different O serotypes isolated in Thailand from 1982 to 1995 were selected and studied for the presence of class 1 integrons, a new group of genetic elements which carry antibiotic resistance genes. Using PCR and DNA...... strains. Serotype O139 strains did not contain class 1 integrons. However, the appearance and disappearance of the O139 serotype in the coastal city Samutsakorn in 1992 and 1993 were associated with the emergence of a distinct V. cholerae O1 strain which contained the aad-V resistance gene cassette. A 150....... cholerae O serotypes of mainly clinical origin in Thailand....

  1. Frequency of Pseudomonas aeruginosa serotypes in burn wound infections and their resistance to antibiotics.

    Estahbanati, Hamid Karimi; Kashani, Parnian Pour; Ghanaatpisheh, Fahimeh


    Pseudomonas aeruginosa plays a prominent role as an etiological agent involved in serious infections in burned patients. In this study P. aeruginosa infections were analyzed at the Motahari Burn Center in Tehran (from 22 December 1998 to April 1999) to estimate their frequency, antibiotic susceptibility and serotypes. One hundred and eighty-four positive cultures and 205 bacterial strains were isolated among swabs or biopsy specimens during the study period. Pseudomonas was found to be the most common (57%) followed by Acinetobacter (17%), Escherichia coli (12%), Staphylococcus aureus (8%) and other organisms (6%). The frequency of P. aeruginosa resistance to gentamicin, ceftizoxime, carbenicillin, cephalothin and ceftazidime was over 90%. The antibiotics to which P. aeruginosa was most sensitive were amikacin and tetracyclin. The "O" serotypes isolated from the 117 Pseudomona aeroginosa isolates were serotypes O:2, O:5, O:6, O:8, O:11, O:12 and O:16. The most common serotype was O:6 (20/17%) followed by O:11 (18/15%) and O:5 (14/12%). The serotype most resistant was O:16 (8%) and the most sensitive was O:8 (2%). Since treatment of infection with available antibiotics according to the results attained proved to be difficult, prevention of infection in the burned patients is considered as an appropriate means of conquering overcoming infection problems. The sum of frequencies of serotypes O:6, O:11, O:5 and O:16 was more than 60%, therefore vaccination of burn patients with polyvalent antiserum to these serotypes could possibly produce immunity in more than half of the burned patients. PMID:12052372

  2. Subtyping of Salmonella Food Isolates Suggests the Geographic Clustering of Serotype Telaviv.

    Durul, Bora; Acar, Sinem; Bulut, Ece; Kyere, Emmanuel O; Soyer, Yeşim


    Salmonella is commonly found in a variety of food products and is a major cause of bacterial foodborne illness throughout the world. In this study, we investigated the prevalence and diversity of Salmonella in eight different food types: sheep ground meat, cow ground meat, chicken meat, cow offal, traditional Sanliurfa cheese, unripened feta cheese, pistachios, and isot (a spice blend of dried red peppers specific to Sanliurfa), traditionally and commonly consumed in Turkey. Among 192 food samples, Salmonella was detected in 59 samples, with the highest prevalence in raw poultry parts (58%) and offal (58%) samples, while Salmonella was not detected in pistachios and dried red pepper. Resultant Salmonella isolates were characterized by serotyping, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). Ten different serotypes represented 10 MLST sequence types (STs) with 1 novel ST and 17 PFGE types. Antimicrobial resistance profiling revealed that 30.5% of the isolates were resistant to two or more antimicrobials. Salmonella enterica subsp. enterica serotype Telaviv, which is rare throughout the world, was the second most common serotype isolated from food samples in this study, suggesting that this serotype might be one of the subtypes that is endemic to Turkey. PMID:26489049

  3. Recurrent Invasive Pneumococcal Disease Serotype 12F in a Vaccinated Splenectomized Patient

    Blaabjerg, Anne Katrine; Schumacher, Anna Holst; Kantsø, Bjørn; Kristensen, Lena Hagelskjær; Schumacher, Helga


    This is the first case report of recurrent invasive pneumococcal disease (IPD), specifically, due to serotype 12F. The patient described here was vaccinated with the 23-valent pneumococcal polysaccharide vaccine (PPV23) due to previous splenectomy, and an anti-pneumococcal IgG test concluded that she had responded sufficiently to vaccination. Still, she had a fulminate recurrent infection with PPV23 serotype 12F. We investigated the anti-pneumococcal IgG test, and it turned out that it is based on the geometric mean value of only 12 of the serotypes included in PPV23; 12F is none of them. The reason is that there are no titer cut-offs available for 11 of the PPV23 serotypes, including 12F, neither nationally nor internationally. Yet, this is not specified in the answer to the clinicians. This case illustrates the need for titer cut-offs for the remaining pneumococcal serotypes in available vaccines, in order to get a more accurate estimation of the vaccination coverage for the individual patient. Therefore, more research on this area is warranted, along with a discussion of whether the laboratory answers to the clinicians should be more detailed.

  4. Benzoquinones as inhibitors of botulinum neurotoxin serotype A.

    Bremer, Paul T; Hixon, Mark S; Janda, Kim D


    Although botulinum neurotoxin serotype A (BoNT/A) is known for its use in cosmetics, it causes a potentially fatal illness, botulism, and can be used as a bioterror weapon. Many compounds have been developed that inhibit the BoNTA zinc-metalloprotease light chain (LC), however, none of these inhibitors have advanced to clinical trials. In this study, a fragment-based approach was implemented to develop novel covalent inhibitors of BoNT/A LC. First, electrophilic fragments were screened against BoNT/A LC, and benzoquinone (BQ) derivatives were found to be active. In kinetic studies, BQ compounds acted as irreversible inhibitors that presumably covalently modify cysteine 165 of BoNT/A LC. Although most BQ derivatives were highly reactive toward glutathione in vitro, a few compounds such as natural product naphthazarin displayed low thiol reactivity and good BoNT/A inhibition. In order to increase the potency of the BQ fragment, computational docking studies were employed to elucidate a scaffold that could bind to sites adjacent to Cys165 while positioning a BQ fragment at Cys165 for covalent modification; 2-amino-N-arylacetamides met these criteria and when linked to BQ displayed at least a 20-fold increase in activity to low μM IC₅₀ values. Unlike BQ alone, the linked-BQ compounds demonstrated only weak irreversible inhibition and therefore acted mainly as non-covalent inhibitors. Further kinetic studies revealed a mutual exclusivity of BQ covalent inactivation and competitive inhibitor binding to sites adjacent to Cys165, refuting the viability of the current strategy for developing more potent irreversible BoNT/A inhibitors. The highlights of this study include the discovery of BQ compounds as irreversible BoNT/A inhibitors and the rational design of low μM IC50 competitive inhibitors that depend on the BQ moiety for activity. PMID:24984937

  5. Effects of crp deletion in Salmonella enterica serotype Gallinarum

    Rubino Salvatore


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

  6. Development of an Indirect Enzyme-Linked Immunosorbent Assay for Seromonitoring Contagious Bovine Pleuropneumonia Using Recombinant Lipoprotein LppQ of Mycoplasma mycoides subsp mycoides SC as Antigen

    XIN Jiu-qing; GAO Yun-long; LI Yuan; WANG Yan-fan; QIAN Ai-dong


    Mycoplasma mycoides subsp mycoides SC (MmmSC) is the etiological agent of contagious bovine pleuropneumonia (CBPP). The lipoprotein LppQ encoded by lppQ gene is specific to MmmSC and is found in the type strain and in field strains isolated in Europe, Africa, and Australia, as well as in vaccine strains. No serological cross-reactions were observed with the related mycoplasmas of the Mycoplasma mycoides cluster. The N-terminal domain of the mature lipoprotein LppQ is hydrophilic, and it induces a strong, specific, early, and persistent immune response in naturally and experimentally infected animals. Mycoplasma-specific TGA (Trp) codons are utilized as stop codons in most other organisms. The lppQ N-terminal fragment from MmmSC HVRI Ⅹ strain, the Chinese strain for CF antigen production, was mutated with one-step overlapping extension PCR. Sequence analysis confirmed the successful mutation from A to G in codon 198 in the lppQ gene. The fragment containing the mutation site was subcloned into the pET32a expression vector. The recombinant protein with molecular weight of 42 kDa was purified using the Ni-NTA His.Bind purification kit, with a purity of up to 95%. Western blot indicated that the standard positive serum of CBPP could react with the recombinant protein. The purified protein was diluted to 0.35 μg mL-1, and coated to microtiter enzyme-linked immunosorbent assay (ELISA) plates. Indirect ELISA reaction conditions were optimized. The value of P/N was determined to be 4.8 (0.934/0.193), the sensitivity to be 95.8% (46/48), and the specificity to be 98.9% (161/163). 3 817 cattle serum samples from three different provinces were detected by the indirect ELISA and CFT. The Kappa value is 0.63, which is middle or high agreement between the two methods.

  7. Serotypes and Clonal Diversity of Streptococcus pneumoniae Causing Invasive Disease in the Era of PCV13 in Catalonia, Spain.

    Eva Del Amo

    Full Text Available The aim of this study was to study the serotypes and clonal diversity of pneumococci causing invasive pneumococcal disease in Catalonia, Spain, in the era of 13-valent pneumococcal conjugate vaccine (PCV13. In our region, this vaccine is only available in the private market and it is estimated a PCV13 vaccine coverage around 55% in children. A total of 1551 pneumococcal invasive isolates received between 2010 and 2013 in the Molecular Microbiology Department at Hospital Sant Joan de Déu, Barcelona, were included. Fifty-two serotypes and 249 clonal types-defined by MLST-were identified. The most common serotypes were serotype 1 (n = 182; 11.7%, 3 (n = 145; 9.3%, 19A (n = 137; 8.8% and 7F (n = 122; 7.9%. Serotype 14 was the third most frequent serotype in children < 2 years (15 of 159 isolates. PCV7 serotypes maintained their proportion along the period of study, 16.6% in 2010 to 13.4% in 2013, whereas there was a significant proportional decrease in PCV13 serotypes, 65.3% in 2010 to 48.9% in 2013 (p<0.01. This decrease was mainly attributable to serotypes 19A and 7F. Serotype 12F achieved the third position in 2013 (n = 22, 6.4%. The most frequent clonal types found were ST306 (n = 154, 9.9%, ST191 (n = 111, 7.2%, ST989 (n = 85, 5.5% and ST180 (n = 80, 5.2%. Despite their decrease, PCV13 serotypes continue to be a major cause of disease in Spain. These results emphasize the need for complete PCV13 vaccination.

  8. Usefulness of real time PCR for the differentiation and quantification of 652 and JP2 Actinobacillus actinomycetemcomitans genotypes in dental plaque and saliva

    Piras Vincenzo


    Full Text Available Abstract Background The aim of our study is to describe a fast molecular method, able to distinguish and quantize the two different genotypes (652 and JP2 of an important periodontal pathogen: Actinobacillus actinomycetemcomitans. The two genotypes show differences in the expression of an important pathogenic factor: the leukotoxin (ltx. In order to evidence this, we performed a real time PCR procedure on the ltx operon, able to recognize Aa clinical isolates with different leukotoxic potentials. Methods The specificity of the method was confirmed in subgingival plaque and saliva specimens collected from eighty-one Italian (Sardinian subjects with a mean age of 43.9, fifty five (68 % of whom had various clinical forms of periodontal disease. Results This procedure showed a good sensitivity and a high linear dynamic range of quantization (107-102 cells/ml for all genotypes and a good correlation factor (R2 = 0.97–0.98. Compared with traditional cultural methods, this real time PCR procedure is more sensitive; in fact in two subgingival plaque and two positive saliva specimens Aa was only detected with the molecular method. Conclusion A low number of Sardinian patients was found positive for Aa infections in the oral cavity, (just 10 positive periodontal cases out of 81 and two of these were also saliva positive. The highly leukotoxic JP2 strain was the most representative (60 % of the positive specimens; the samples from periodontal pockets and from saliva showed some ltx genotype for the same patient. Our experience suggests that this approach is suitable for a rapid and complete laboratory diagnosis for Aa infection.

  9. Improved Multiplex PCR Using Conserved and Species-Specific 16S rRNA Gene Primers for Simultaneous Detection of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis

    Tran, Simon Dangtuan; Rudney, Joel D.


    Among putative periodontal pathogens, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis are most convincingly implicated as etiological agents in periodontitis. Therefore, techniques for detection of those three species would be of value. We previously published a description of a multiplex PCR that detects A. actinomycetemcomitans and P. gingivalis. The present paper presents an improvement on that technique, which now allows more sensitive detection of all three periodontal pathogens. Sensitivity was determined by testing serial dilutions of A. actinomycetemcomitans, B. forsythus, and P. gingivalis cells. Primer specificity was tested against (i) all gene sequences from the GenBank-EMBL database, (ii) six A. actinomycetemcomitans, one B. forsythus, and four P. gingivalis strains, (iii) eight different species of oral bacteria, and (iv) supra- and subgingival plaque samples from 20 healthy subjects and subgingival plaque samples from 10 patients with periodontitis. The multiplex PCR had a detection limit of 10 A. actinomycetemcomitans, 10 P. gingivalis, and 100 B. forsythus cells. Specificity was confirmed by the fact that (i) none of our forward primers were homologous to the 16S rRNA genes of other oral species, (ii) amplicons of predicted size were detected for all A. actinomycetemcomitans, B. forsythus, and P. gingivalis strains tested, and (iii) no amplicons were detected for the eight other bacterial species. A. actinomycetemcomitans, B. forsythus, and P. gingivalis were detected in 6 of 20, 1 of 20, and 11 of 20 of supragingival plaque samples, respectively, and 4 of 20, 7 of 20, and 13 of 20 of subgingival plaque samples, respectively, from periodontally healthy subjects. Among patients with periodontitis, the organisms were detected in 7 of 10, 10 of 10, and 7 of 10 samples, respectively. The simultaneous detection of three periodontal pathogens is an advantage of this technique over conventional PCR assays. PMID

  10. Neural Network Model for Survival and Growth of Salmonella enterica Serotype 8,20:-:z6 in Ground Chicken Thigh Meat during Cold Storage: Extrapolation to Other Serotypes.

    Oscar, T P


    Mathematical models that predict the behavior of human bacterial pathogens in food are valuable tools for assessing and managing this risk to public health. A study was undertaken to develop a model for predicting the behavior of Salmonella enterica serotype 8,20:-:z6 in chicken meat during cold storage and to determine how well the model would predict the behavior of other serotypes of Salmonella stored under the same conditions. To develop the model, ground chicken thigh meat (0.75 cm(3)) was inoculated with 1.7 log Salmonella 8,20:-:z6 and then stored for 0 to 8 -8 to 16°C. An automated miniaturized most-probable-number (MPN) method was developed and used for the enumeration of Salmonella. Commercial software (Excel and the add-in program NeuralTools) was used to develop a multilayer feedforward neural network model with one hidden layer of two nodes. The performance of the model was evaluated using the acceptable prediction zone (APZ) method. The number of Salmonella in ground chicken thigh meat stayed the same (P > 0.05) during 8 days of storage at -8 to 8°C but increased (P < 0.05) during storage at 9°C (+0.6 log) to 16°C (+5.1 log). The proportion of residual values (observed minus predicted values) in an APZ (pAPZ) from -1 log (fail-safe) to 0.5 log (fail-dangerous) was 0.939 for the data (n = 426 log MPN values) used in the development of the model. The model had a pAPZ of 0.944 or 0.954 when it was extrapolated to test data (n = 108 log MPN per serotype) for other serotypes (S. enterica serotype Typhimurium var 5-, Kentucky, Typhimurium, and Thompson) of Salmonella in ground chicken thigh meat stored for 0 to 8 days at -4, 4, 12, or 16°C under the same experimental conditions. A pAPZ of ≥0.7 indicates that a model provides predictions with acceptable bias and accuracy. Thus, the results indicated that the model provided valid predictions of the survival and growth of Salmonella 8,20:-:z6 in ground chicken thigh meat stored for 0 to 8 days at -8 to

  11. Does age acquired immunity confer selective protection to common serotypes of Campylobacter jejuni?

    Ogden Iain D


    Full Text Available Abstract Background Campylobacter infection is a major cause of bacterial gastrointestinal disease. Exposure to Campylobacter is known to produce an immune response in humans that can prevent future symptomatic infections. Further, studies of the general population have shown that seroprevalence to Campylobacter increases with age. Methods A large collection of serotyped Campylobacter isolates, obtained from human clinical faecal samples, were analysed by comparing the ratio of uncommon to common serotypes by different age groups, using χ2 tests. Results We have identified that older age groups, as well as having generally lower incidence, are significantly less likely to be infected by the more common serotypes. Conclusion These results are indicative of acquired immunity, however, further studies are needed to rule out the confounding effects of the variations in exposure pathways experienced by different age groups.

  12. Simultaneous circulation of all four dengue serotypes in Manaus, State of Amazonas, Brazil in 2011

    Michele de Souza Bastos


    Full Text Available INTRODUCTION: Manaus, the capital city of the state of Amazon with nearly 2 million inhabitants, is located in the middle of the Amazon rain forest and has suffered dengue outbreaks since 1998. METHODS: In this study, blood samples were investigated using reverse transcriptase-polymerase chain reaction (RT-PCR, aimed at identifying dengue virus serotypes. RESULTS: Acute phase sera from 432 patients were tested for the presence of dengue virus. Out of the 432 patients, 137 (31.3% were found to be positive. All the four dengue virus serotypes were observed. CONCLUSIONS: The simultaneous circulation of the four dengue serotypes is described for the first time in Manaus and in Brazil.

  13. Effect of Culicoides sonorensis salivary proteins on clinical disease outcome in experimental Bleutongue virus serotype 8 infection of Dorset sheep

    Drolet, B.S.; Reister, L.M.; Lehiy, C.J.; Rijn, van P.A.; Bowen, R.A.


    The severity of Bluetongue clinical disease in ruminants varies greatly depending on the outbreak serotype/strain, animal species/breed, and immune status of the herd. To predict disease risk from any of the 26 Bluetongue virus (BTV) serotypes identified to date, experimental animal susceptibility s

  14. Adenoviral-based foot-and-mouth disease virus vaccine: evaluation of new vectors expressing serotype O in bovines

    Foot-and-mouth disease virus (FMDV), an antigenically variable virus, is considered the most important infectious disease of cloven-hoofed animals. Recently serotypes A and O have been the cause of major outbreaks. We previously demonstrated that an adenovirus-based FMDV serotype A24 subunit vaccine...

  15. A simulated metagenomic approach for bacterial serotyping using shotgun genome sequences coupled with O-Antigen gene cluster analysis

    Background: Accurate determination of food-borne pathogen serotype and genotype information is important for disease surveillance and outbreak source tracking. E. coli serotype O157:H7 and non-O157 of Shiga toxin-producing E. coli (STEC) serogroups, including O26, O45, O103, O111, O121, O145 (top ...

  16. Streptococcus pneumoniae from Palestinian nasopharyngeal carriers: serotype distribution and antimicrobial resistance.

    Abedelmajeed Nasereddin

    Full Text Available Infections of Streptococcus pneumoniae in children can be prevented by vaccination; left untreated, they cause high morbidity and fatalities. This study aimed at determining the nasopharyngeal carrier rates, serotype distribution and antimicrobial resistance patterns of S. pneumoniae in healthy Palestinian children under age two prior to the full introduction of the pneumococcal 7-valent conjugate vaccine (PCV7, which was originally introduced into Palestine in a pilot trial in September, 2010. In a cross sectional study, nasopharyngeal specimens were collected from 397 healthy children from different Palestinian districts between the beginning of November 2012 to the end of January 2013. Samples were inoculated into blood agar and suspected colonies were examined by amplifying the pneumococcal-specific autolysin gene using a real-time PCR. Serotypes were identified by a PCR that incorporated different sets of specific primers. Antimicrobial susceptibility was measured by disk diffusion and MIC methods. The resulting carrier rate of Streptococcus pneumoniae was 55.7% (221/397. The main serotypes were PCV7 serotypes 19F (12.2%, 23F (9.0%, 6B (8.6% and 14 (4% and PCV13 serotypes 6A (13.6% and 19A (4.1%. Notably, serotype 6A, not included in the pilot trial (PCV7 vaccine, was the most prevalent. Resistance to more than two drugs was observed for bacteria from 34.1% of the children (72/211 while 22.3% (47/211 carried bacteria were susceptible to all tested antibiotics. All the isolates were sensitive to cefotaxime and vancomycin. Any or all of these might impinge on the type and efficacy of the pneumococcal conjugate vaccines and antibiotics to be used for prevention and treatment of pneumococcal disease in the country.

  17. Identification of the serotypes of bacterial meningitis agents; implication for vaccine usage.

    Mohammad Mehdi Attarpour-Yazdi


    Full Text Available Bacterial meningitis is one of the most serious infections and should be treated as emergency. As it has significant morbidity and mortality throughout the world, every country should have precise information regarding the etiological agents of disease and populations at risk to design public health prevention strategy. In the present study in addition of evaluation of common etiological agents (Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae in bacterial meningitis cases, we sero-grouped or serotyped the obtained agents in order to predict the usefulness of existing vaccines against bacterial meningitis.Cerebrospinal fluid of 182 suspected meningitis patients were collected, from which 114 cases were approved by biochemical, microbiological and molecular tests as bacterial meningitis. The isolated bacteria were serogrouped or serotyped to determine the dominant serotypes.Streptococcus pneumoniae accounted for 36%, Haemophilus influenza for 26% and Neisseria meningitidis for 14% of cases. From 13 serogroups of N. meningitides the most frequent serogroups, were meningococcus group B (51%, C(24% A (18%, Z(2%, W135 (1% and 3% was not identified. In H. influenzae group only serotype b (100% have been identified and in pneumococcal meningitis the most common serotype among our cases were 18C (44% followed by14 (17%, 19A (13%, 6A (9%, 7F (4%, 4(3%, 3 (3%, 9V (2%, 8 (2%, 23f (2%, 5 (1%.Since there is no nationwide mass immunization program for common agents of bacterial meningitis in Iran, the result of this study can be used to improve the existing vaccines to cover the detected serotypes and consequently reduce the incidence of bacterial meningitis.

  18. Pediatric invasive pneumococcal disease caused by vaccine serotypes following the introduction of conjugate vaccination in Denmark.

    Zitta B Harboe

    Full Text Available A seven-valent pneumococcal conjugate vaccine (PCV7 was introduced in the Danish childhood immunization program (2+1 schedule in October 2007, followed by PCV13 starting from April 2010. The nationwide incidence of IPD among children younger than 5 years nearly halved after the introduction of PCV7 in the program, mainly due to a decline in IPD caused by PCV7-serotypes. We report the results from a nationwide population-based cohort study of laboratory confirmed IPD cases in children younger than 5 years during October 1, 2007 to December 31, 2010 and describe the characteristics of children suspected to present with a vaccine failure. The period between April 19 and December 31, 2010 was considered a PCV7/PCV13 transitional period, where both vaccines were offered. We identified 45 episodes of IPD caused by a PCV7 serotype (23% of the total number and 105 (55% caused by one of the 6 additional serotypes in PCV13. Ten children had received at least one PCV7 dose before the onset of IPD caused by a PCV7 serotype. Seven children were considered to be incompletely vaccinated before IPD, but only three cases fulfilled the criteria of vaccine failure (caused by serotypes 14, 19F and 23F. One case of vaccine failure was observed in a severely immunosuppressed child following three PCV7 doses, and two cases were observed in immunocompetent children following two infant doses before they were eligible for their booster. None of the IPD cases caused by the additional PCV13 serotypes had been vaccinated by PCV13 and there were therefore no PCV13-vaccine failures in the first 8-months after PCV13 introduction in Denmark.

  19. Association of Serotype-Specific Antibody Concentrations and Functional Antibody Titers with Subsequent Pneumococcal Carriage in Toddlers Immunized with a 9-Valent Pneumococcal Conjugate Vaccine

    Simell, Birgit; Nurkka, Anu; Lahdenkari, Mika; Givon-Lavi, Noga; Käyhty, Helena; Dagan, Ron; Jokinen, Jukka


    Association of pneumococcal nasopharyngeal carriage with the concentration and opsonophagocytic activity (OPA) of serum serotype-specific antibodies was determined for toddlers 1 month after immunization with a 9-valent pneumococcal conjugate vaccine. Higher anti-serotype 14 and anti-serotype 19F IgG and anti-serotype 14 IgM correlated with a lowered probability of pneumococcal acquisition. Postvaccination OPA did not correlate with pneumococcal carriage.

  20. Air Travel Is Associated with Intracontinental Spread of Dengue Virus Serotypes 1-3 in Brazil

    Nunes, Marcio R. T.; Palacios, Gustavo; Faria, Nuno Rodrigues; Sousa, Edivaldo Costa; Pantoja, Jamilla A; Rodrigues, Sueli G.; Carvalho, Valéria L.; Medeiros, Daniele B A; Savji, Nazir; Baele, Guy; Suchard, Marc A.; Lemey, Philippe; Pedro F. C. Vasconcelos; Lipkin, W. Ian


    Dengue virus and its four serotypes (DENV-1 to DENV-4) infect 390 million people and are implicated in at least 25,000 deaths annually, with the largest disease burden in tropical and subtropical regions. We investigated the spatial dynamics of DENV-1, DENV-2 and DENV-3 in Brazil by applying a statistical framework to complete genome sequences. For all three serotypes, we estimated that the introduction of new lineages occurred within 7 to 10-year intervals. New lineages were most likely to b...

  1. Genetic Analysis of the Capsule Locus of Haemophilus influenzae Serotype f

    Satola, Sarah W.; Patricia L Schirmer; Farley, Monica M.


    A 19-kb DNA region containing genes involved in the biosynthesis of the capsule of Haemophilus influenzae serotype f (Hif) has been cloned and characterized. The Hif cap locus organization is typical of group II capsule biosynthetic loci found in other H. influenzae serotype b bacteria and other gram-negative bacteria. However, the Hif cap locus was not associated with an IS1016 element. Three new open reading frames, Fcs1, Fcs2, and Fcs3, were identified in the Hif capsule-specific region II...

  2. Distribution of Serotypes and Antimicrobial Resistance Genes among Streptococcus agalactiae Isolates from Bovine and Human Hosts

    Dogan, Belgin; Schukken, Y. H.; de Santisteban, C.; Boor, Kathryn J.


    To better understand the emergence and transmission of antibiotic-resistant Streptococcus agalactiae, we compared phenotypic and genotypic characteristics of 52 human and 83 bovine S. agalactiae isolates. Serotypes found among isolates from human hosts included V (48.1%), III (19.2%), Ia and Ib (13.5% each), and II (5.8%). Among isolates from bovine hosts, molecular serotypes III and II were predominant (53 and 14.5%, respectively). Four and 21 different ribotypes were found among human and b...

  3. Outer membrane protein and biotype analysis of non-serotypable strains of Haemophilus influenzae.

    Coverdale, C H; Temple, G S


    Strains of Haemophilus influenzae (n = 161) were isolated from inpatients with symptoms of pulmonary infection. Conventional tests showed that 144 strains were non-serotypable and all belonged to one of eight biotypes. The common biotypes were 2 (41%), 3 (27.1%), 1 (13.2%) and 5 (10.4%). The outer membrane protein (OMP) profiles of 59 non-serotypable strains were examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). A comparison of OMP profiles suggested a possibl...

  4. Genome-wide dissection of globally emergent multi-drug resistant serotype 19A Streptococcus pneumoniae

    Dylan R Pillai; Shahinas, Dea; Buzina, Alla; Pollock, Remy A; Lau, Rachel; Khairnar, Krishna; Wong, Andrew; Farrell, David J.; Green, Karen; McGeer, Allison; Low, Donald E.


    Background Emergence of multi-drug resistant (MDR) serotype 19A Streptococcus pneumoniae (SPN) is well-documented but causal factors remain unclear. Canadian SPN isolates (1993-2008, n = 11,083) were serotyped and in vitro susceptibility tested. A subset of MDR 19A were multi-locus sequence typed (MLST) and representative isolates' whole genomes sequenced. Results MDR 19A increased in the post-PCV7 era while 19F, 6B, and 23F concurrently declined. MLST of MDR 19A (n = 97) revealed that sequen...

  5. Serotyping, Genotyping, and Antimicrobial Susceptibility of Ornithobacterium rhinotracheale Isolates from Mexico.

    Peña-Vargas, Edgar Rafael; Vega-Sánchez, Vicente; Morales-Erasto, Vladimir; Trujillo-Ruíz, Héctor Hugo; Talavera-Rojas, Martín; Soriano-Vargas, Edgardo


    The bacterium Ornithobacterium rhinotracheale is associated with respiratory disease and septicemia in poultry. In this study, 9 reference strains and a total of 23 isolates of O. rhinotracheale from respiratory diseased poultry from Mexico were serotyped and genotyped. Furthermore, the antimicrobial susceptibility of isolates and reference strains of O. rhinotracheale were determined. All isolates belong to serotype A and showed a clonal relationship. All reference strains and isolates were resistant to colistin, fosfomycin, gentamicin, kanamycin, streptomycin, and trimethoprim-sulfamethoxazole. These results should eventually be helpful in planning strategies for the control of O. rhinotracheale infections in poultry in Mexico. PMID:27610728


    Made Agus Hendrayana; Komang Januartha Putra Pinatih; Amy Yelly


    Escherichia coli (E.coli)  is the pathogenic bacteria which commonly associated with intestine and kidney infection in human. One of the strain of zoonotic E.coli serotype O157. Human and cattle, mostly swine are predominant reservoirs for E. coli O157. Swine butchering in Bali specifically in Denpasar is increasing each year. The aim of this study was to identify E.coli serotype O157 in pork in Denpasar. They were 31 samples of pork taken from different butchers in 24 traditional markets and...

  7. Serotype-Specific Cell-Mediated Immunity Associated With Clearance of Homotypic Group B Streptococcus Rectovaginal Colonization in Pregnant Women.

    Kwatra, Gaurav; Adrian, Peter V; Shiri, Tinevimbo; Izu, Alane; Cutland, Clare L; Buchmann, Eckhart J; Madhi, Shabir A


    We investigated the association between group B Streptococcus (GBS) serotype-specific capsular polysaccharide cellular immunity, measured with enzyme-linked immunospot (ELISPOT) interferon γ release assay at 20 weeks gestation in pregnant women, and its effect on rectovaginal serotype-specific GBS colonization up to 37 weeks gestation. Among women colonized by serotype III at enrollment, interferon γ ELISPOT positivity was more common in those in whom colonization was cleared (44.4%) than in those in whom colonization persisted (7.4%; P = .008), with a similar trend observed for serotype Ia. Presence of serotype-specific capsular polysaccharide cell-mediated immunity contributes to the clearance of GBS rectovaginal colonization. PMID:27029777

  8. Validation and characterization of a human volunteer challenge model for cholera by using frozen bacteria of the new Vibrio cholerae epidemic serotype, O139

    Cohen, MB; Giannella, RA; Losonsky, GA; Lang, DR; Parker, S; Hawkins, JA; Gunther, C; Schiff, GA


    Until recently, all epidemic strains of Vibrio cholerae were of the O1 serotype. Current epidemics have also been caused by a new serotype, Vibrio cholerae O139. Although the pathogenesis and clinical features of O139 cholera are similar to those of O1 cholera, immunity to serotype O1 does not confe

  9. The use of serotype 1-and serotype 3-specific polymerase chain reaction for the detection of Marek's disease virus in chickens

    Handberg, Kurt; Nielsen, Ole L.; Jørgensen, Poul Henrik


    to develop and evaluate a reliable and easy-to-handle method for surveillance of the occurrence of MDV in chicken flocks. We emphasize the development of a method, which can be applied to types of samples conveniently collected in the field, e.g. feather tips and blood samples. In addition, the PCR...... albumen pretreatment. The PCR proved to be a convenient tool for the monitoring of MDV in the poultry population, and feather tips were the most convenient and sensitive samples.......A serotype 1- and serotype 3-specific detection of Marek's disease virus (MDV) by polymerase chain reaction (PCR) was developed. The sensitivity of the method when applied to cell culture grown virus was comparable with that of cultivation. The method was applied to various tissue samples from...

  10. Distribution of Campylobacter jejuni Penner serotypes in broiler flocks 1998-2000 in a small Danish community with special reference to serotype 4-complex

    Wedderkopp, A.; Nielsen, E.M.; Pedersen, Karl


    During the period January 1998-December 2001, all Danish broiler flocks were monitored bacteriologically for thermophilic campylobacters and isolates were stored at -80 degreesC. Six neighbouring broiler farms in a small community were selected for detailed examination of all Campylobacter jejuni...... isolated (n = 180) from these farms during 1998-2000 using Penner serotyping and pulsed-field gel electrophoresis (PFGE). The area and the farms were selected according to their prevalence of campylobacter so that both farms with low and high frequencies of campylobacter positive flocks were included in...... the study. The frequency of campylobacter positive flocks on the six farms ranged from 24.5 to 72.7%. One hundred and eighty of the isolates were C. jejuni (included in this study), 14 isolates were C. coli whereas 7 isolates belonged to other species but were not further identified. By serotyping of...

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

    Laetitia Fabre

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

  12. Antibodies Directed against Shiga-Toxin Producing Escherichia coli Serotype O103 Type III Secreted Proteins Block Adherence of Heterologous STEC Serotypes to HEp-2 Cells.

    Taseen S Desin

    Full Text Available Shiga toxin-producing Escherichia coli (STEC serotype O103 is a zoonotic pathogen that is capable of causing hemorrhagic colitis and hemolytic uremic syndrome (HUS in humans. The main animal reservoir for STEC is ruminants and hence reducing the levels of this pathogen in cattle could ultimately lower the risk of STEC infection in humans. During the process of infection, STECO103 uses a Type III Secretion System (T3SS to secrete effector proteins (T3SPs that result in the formation of attaching and effacing (A/E lesions. Vaccination of cattle with STEC serotype O157 T3SPs has previously been shown to be effective in reducing shedding of STECO157 in a serotype-specific manner. In this study, we tested the ability of rabbit polyclonal sera against individual STECO103 T3SPs to block adherence of the organism to HEp-2 cells. Our results demonstrate that pooled sera against EspA, EspB, EspF, NleA and Tir significantly lowered the adherence of STECO103 relative to pre-immune sera. Likewise, pooled anti-STECO103 sera were also able to block adherence by STECO157. Vaccination of mice with STECO103 recombinant proteins induced strong IgG antibody responses against EspA, EspB, NleA and Tir but not against EspF. However, the vaccine did not affect fecal shedding of STECO103 compared to the PBS vaccinated group over the duration of the experiment. Cross reactivity studies using sera against STECO103 recombinant proteins revealed a high degree of cross reactivity with STECO26 and STECO111 proteins implying that sera against STECO103 proteins could potentially provide neutralization of attachment to epithelial cells by heterologous STEC serotypes.

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


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

  14. Serotypes and typability of Campylobacter jejuni and Campylobacter coli isolated from poultry products

    Nielsen, Eva Møller; Nielsen, Niels Ladefoged


    Campylobacter infection is one of the most common bacterial enteric pathogens. Campylobacter jejuni and Campylobacter coli infections are mostly food- and waterborne and especially poultry is often assumed to be an important source. The heat-stable serotyping system (the 'Penner' scheme) was used...

  15. Epidemiologic Investigation of Riemerella anatipestifer in a Commercial Duck Company by Serotyping and DNA Fingerprinting

    A commercial duck company that raises approximately two million Pekin ducks per year experienced an outbreak of Riemerella anatipestifer(RA)on nine farms over a one year period. Due to concerns that the bacteria was being spread from farm to farm, an investigation using serotyping and DNA fingerprin...

  16. Passive immunization of pigs against experimental infection with Streptococcus suis serotype 2

    Andresen, Lars Ole; Tegtmeier, Conny


    The safety and protective efficacy of a horse antiserum raised against inactivated whole cell preparations of Streptococcus suis serotype 2 was investigated in pigs by experimental challenge. The antiserum was evaluated in two similar experiments each comprising 12 4-week-old pigs treated with 6 ml...

  17. Foot-and-mouth disease virus serotype SAT 3 in long-horned ankole calf, Uganda

    Dhikusooka, Moses Tefula; Tjørnehøj, Kirsten; Ayebazibwe, Chrisostom; Namatovu, Alice; Ruhweza, Simon; Siegismund, Hans Redlef; Wekesa, Sabenzia Nabalayo; Normann, Preben; Belsham, Graham J.


    After a 16-year interval, foot-and-mouth disease virus serotype SAT 3 was isolated in 2013 from an apparently healthy long-horned Ankole calf that grazed close to buffalo in Uganda. The emergent virus strain is ≈20% different in nucleotide sequence (encoding VP1 [viral protein 1]) from its closest relatives isolated previously from buffalo in Uganda.

  18. Twinned crystals of adeno-associated virus serotype 3b prove suitable for structural studies

    Lerch, Thomas F.; Xie, Qing; Ongley, Heather M.; Hare, Joan; Chapman, Michael S.


    Crystals of adeno-associated virus serotype 3b, a human DNA virus with promise as a vector for gene therapy, have been grown, diffract X-rays to ∼2.6 Å resolution and are suitable for structure determination in spite of twinning.

  19. Rapid Molecular Determination of Serotype from Clinical Isolates of Salmonella Enterica

    Background: The conventional serotyping of Salmonella Enterica is time consuming, costly, and requires highly skilled staff. In the present study, we report a multiplex PCR typing method using capillary electrophoresis for fragment analysis that allows for the identification of the 30 most common h...

  20. Serotype distribution and antimicrobial resistance of Streptococcus pneumoniae causing noninvasive diseases in a Children's Hospital, Shanghai

    Fen Pan


    Full Text Available Background:Streptococcus pneumoniae, which cause noninvasive pneumococcal diseases, severely impair children's health. This study analyzed serotype distribution and antimicrobial resistance of S. pneumoniae from January 2012 to December 2012 in a Children's Hospital, Shanghai.Methods:A total of 328 pneumococcal isolates were serotyped by multiplex sequential PCR and/or capsule-quellung reaction. The minimum inhibitory concentrations for 11 antimi- crobial agents were determined by broth microdilution method.Results:Among 328 strains, 19F (36.3%, 19A (13.4%, 6A (11.9%, 23F (11.0%, 14 (5.8%, 6B (5.2%, and 15B/C (4.3% were the most common serotypes. The coverage rates of 7-, 10-, and 13-valent conjugate vaccines (PCV7, PCV10, and PCV13 were 58.2%, 58.2%, and 84.1%, respectively. Out of the isolates, 26 (7.9% strains were penicillin resistant. Most of the strains displayed high resistance rate to macrolides (98.5% to erythromycin, 97.9% to azithromycin, and 97.0% to clindamycin.Conclusions:The potential coverage of PCV13 is higher than PCV7 and PCV10 because of the emergence of 19A and there should be long-term and systematic surveillance for non-vaccine serotypes.

  1. Foot-and-Mouth Disease Virus Serotype SAT 3 in Long-Horned Ankole Calf, Uganda

    Dhikusooka, Moses Tefula; Tjørnehøj, Kirsten; Ayebazibwe, Chrisostom;


    After a 16-year interval, foot-and-mouth disease virus serotype SAT 3 was isolated in 2013 from an apparently healthy long-horned Ankole calf that grazed close to buffalo in Uganda. The emergent virus strain is ≈20% different in nucleotide sequence (encoding VP1 [viral protein 1]) from its closest...

  2. Rapid and early detection of salmonella serotypes with hyperspectral microscope and multivariate data analysis

    This study was designed to evaluate hyperspectral microscope images for early and rapid detection of Salmonella serotypes: S. Enteritidis, S. Heidelberg, S. Infantis, S. Kentucky, and S. Typhimurium at incubation times of 6, 8, 10, 12, and 24 hours. Images were collected by an acousto-optical tunab...

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

    Ktsoyan, Zhanna A.; Mkrtchyan, Mkhitar S.; Zakharyan, Magdalina K.; Mnatsakanyan, Armine A.; Arakelova, Karine A.; Gevorgyan, Zaruhi U.; Ktsoyan, Lusntag A.; Sedrakyan, Anahit; Hovhannisyan, Alvard I; Ghazaryan, Karine A.; Boyajyan, Anna S.; Aminov, Rustam


    The main goal of this study was to establish how the inflammation caused by infection with two different Salmonella enterica serotypes, S. Typhimurium and S. Enteritidis, may lead to the predisposition to allergy as measured by total IgE level in the blood. Infection by S. Typhimurium did not aff...

  4. Serotyping of Toxoplasma gondii in Cats (Felis domesticus) Reveals Predominance of Type II Infections in Germany

    Background: Cats are definitive hosts of Toxoplasma gondii and play an essential role in the epidemiology of this parasite. The study aims at clarifying whether cats are able to develop specific antibodies against different clonal types of T. gondii and to determine by serotyping the T. gondii clona...

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

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


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

  6. Whole-Genome Sequence of Aeromonas hydrophila Strain AH-1 (Serotype O11).

    Forn-Cuní, Gabriel; Tomás, Juan M; Merino, Susana


    Aeromonas hydrophila is an emerging pathogen of aquatic and terrestrial animals, including humans. Here, we report the whole-genome sequence of the septicemic A. hydrophila AH-1 strain, belonging to the serotype O11, and the first mesophilic Aeromonas with surface layer (S-layer) to be sequenced. PMID:27587829

  7. Invasiveness of Serotypes and Clones of Streptococcus pneumoniae among Children in Finland

    William P Hanage; Kaijalainen, Tarja H.; Ritva K Syrjänen; Auranen, Kari; Leinonen, Maija; Mäkelä, P. Helena; Brian G. Spratt


    Streptococcus pneumoniae (the pneumococcus) causes diseases from otitis media to life-threatening invasive infection. The species is extremely antigenically and clonally diverse. We wished to determine odds ratios (ORs) for serotypes and clones of S. pneumoniae that cause invasive disease in Finland. A total of 224 isolates of S. pneumoniae from cases of invasive disease in children

  8. K2 Serotype Klebsiella pneumoniae Causing a Liver Abscess Associated with Infective Endocarditis▿

    Rivero, Andres; Gomez, Eric; Alland, David; David B. Huang; Chiang, Tom


    Klebsiella pneumoniae primary liver abscess (KPLA) is an emerging disease that is associated with distant septic complications. We report the first case of KPLA associated with infective endocarditis. The K. pneumoniae strain was a hypermucoid K2 serotype carrying the rmpA virulence-associated gene.

  9. AAV Natural Infection Induces Broad Cross-Neutralizing Antibody Responses to Multiple AAV Serotypes in Chimpanzees.

    Calcedo, Roberto; Wilson, James M


    Cross-sectional studies of primates have revealed that natural neutralizing antibody (NAb) responses to adeno-associated viruses (AAV) span multiple serotypes. This differs from the phenotype of the NAb response to an AAV vector delivered to seronegative nonhuman primates that is typically restricted to the administered AAV serotype. To better understand the mechanism by which natural AAV infections result in broad NAb responses, we conducted a longitudinal study spanning 10 years in which we evaluated serum-circulating AAV NAb levels in captive-housed chimpanzees. In a cohort of 25 chimpanzees we identified 3 distinct groups of animals: those that never seroconverted to AAV (naïve), those that were persistently seropositive (chronic), and those that seroconverted during the 10-year period (acute). For the chronic group we found a broad seroresponse characterized by NAbs reacting to multiple AAV serotypes. A similar cross-neutralization pattern of NAbs was observed in the acute group. These data support our hypothesis that a single natural infection with AAV induces a broadly cross-reactive NAb response to multiple AAV serotypes. PMID:27314914

  10. Clonal expansion within pneumococcal serotype 6C after use of seven-valent vaccine.

    Nicholas J Loman

    Full Text Available Streptococcus pneumoniae causes invasive infections, primarily at the extremes of life. A seven-valent conjugate vaccine (PCV7 is used to protect against invasive pneumococcal disease in children. Within three years of PCV7 introduction, we observed a fourfold increase in serotype 6C carriage, predominantly due to a single clone. We determined the whole-genome sequences of nineteen S. pneumoniae serotype 6C isolates, from both carriage (n = 15 and disease (n = 4 states, to investigate the emergence of serotype 6C in our population, focusing on a single multi-locus sequence type (MLST clonal complex 395 (CC395. A phylogenetic network was constructed to identify different lineages, followed by analysis of variability in gene sets and sequences. Serotype 6C isolates from this single geographical site fell into four broad phylogenetically distinct lineages. Variation was seen in the 6C capsular locus and in sequences of genes encoding surface proteins. The largest clonal complex was characterised by the presence of lantibiotic synthesis locus. In our population, the 6C capsular locus has been introduced into multiple lineages by independent capsular switching events. However, rapid clonal expansion has occurred within a single MLST clonal complex. Worryingly, plasticity exists within current and potential vaccine-associated loci, a consideration for future vaccine use, target selection and design.

  11. An easy method for detection of nasopharyngeal carriage of multiple Streptococcus pneumoniae serotypes

    Kaltoft, Margit S.; Sørensen, Uffe; Slotved, Hans-Christian;


    In this paper, a simplified method for detection of pneumococcal carriage and for revealing the presence of several serotypes in a nasopharyngeal sample is evaluated. Enrichment broth was used for transportation and for the initial culturing of samples. All specimens were examined directly by the...

  12. Vector competence of Culicoides sonorensis (Diptera: Ceratopogonidae) to epizootic hemorrhagic disease virus serotype 7

    Background: Culicoides sonorensis (Diptera: Ceratopogonidae) is a vector of epizootic hemorrhagic disease virus (EHDV) serotypes 1 and 2 in North America, where these viruses are well-known pathogens of white-tailed deer (WTD) and other wild ruminants. Although historically rare, reports of clinica...

  13. Genomic Epidemiology of Salmonella enterica Serotype Enteritidis based on Population Structure of Prevalent Lineages

    Deng, Xiangyu; Desai, Prerak T.; den Bakker, Henk C.;


    Salmonella enterica serotype Enteritidis is one of the most commonly reported causes of human salmonellosis. Its low genetic diversity, measured by fingerprinting methods, has made subtyping a challenge. We used whole-genome sequencing to characterize 125 S. enterica Enteritidis and 3 S. enterica...

  14. Streptococcus agalactiae Serotype Distribution and Antimicrobial Susceptibility in Pregnant Women in Gabon, Central Africa

    Sabine Belard; Nicole Toepfner; Mesküre Capan-Melser; Ghyslain Mombo-Ngoma; Rella Zoleko-Manego; Mirjam Groger; Pierre-Blaise Matsiegui; Agnandji, Selidji T; Adegnika, Ayôla A.; Raquel González; Kremsner, Peter G; Clara Menendez; Michael Ramharter; Reinhard Berner


    Neonatal invasive disease due to Streptococcus agalactiae is life threatening and preventive strategies suitable for resource limited settings are urgently needed. Protective coverage of vaccine candidates based on capsular epitopes will relate to local epidemiology of S. agalactiae serotypes and successful management of critical infections depends on timely therapy with effective...

  15. Correlation of disease spectrum among four Dengue serotypes: a five years hospital based study from India

    Rajni Kumaria


    Full Text Available The recognition of DF (DHF Dengue Hemorrhagic Fever is very complicated due to occurrence of a wide spectrum of clinical signs and symptoms during acute phase of illness. Moreover, presence of four serotypes further complicates the prognosis. To investigate the predictors of disease severity and elucidate the prognostic markers among four dengue serotypes, this study was conducted on 320 inpatients having acute febrile illness clinically suspected as DI, over a period of five years. Dengue serotypes were confirmed by multiplex reverse transcriptase (RT-PCR. Eighty patients were positive for DI with presence of Den-1, Den-2, Den-3, and Den-4 in 8, 35, 27 and 10 patients, respectively. The severe clinical manifestations, abdominal pain and hepatomegaly, were comparatively higher in Den-2 patients. Liver aminotransferases levels were also higher in Den-2 patients (app. 5 fold. This study clearly indicates the hyperendemicity of all dengue serotypes. Nucleotide sequencing of Envelope region revealed that the presently emerged Den-3 belongs to type III, having high homology with genotype responsible for number of outbreaks in 1980s. The re-emergence of this deadly type can be suspected to cause more outbreaks in future and is a matter of great concern.

  16. Co-circulation of two extremely divergent serotype SAT 2 lineages in Kenya highlights challenges to foot-and-mouth disease control

    Sangula, Abraham; Belsham, Graham; Muwanika, Vincent;


    Amongst the SAT serotypes of foot-and-mouth disease virus (FMDV), the SAT 2 serotype is the most widely distributed throughout sub-Saharan Africa. Kenyan serotype SAT 2 viruses have been reported to display the highest genetic diversity for the serotype globally. This complicates diagnosis...... and control, and it is essential that patterns of virus circulation are known in order to overcome these difficulties. This study was undertaken to establish patterns of evolution of FMDV serotype SAT 2 in Kenya using complete VP1 coding sequences in a dataset of 65 sequences from Africa, collected over...

  17. The concentration of apolipoprotein A-I decreases during experimentally induced acute-phase processes in pigs

    Carpintero, R.; Pineiro, M.; Andres, M.;


    In this work, apolipoprotein A-I (ApoA-I) was purified from pig sera. The responses of this protein after sterile inflammation and in animals infected with Actinobacillus pleuropneumoniae or Streptococcus suis were investigated. Decreases in the concentrations of ApoA-I, two to five times lower...

  18. Knoflook remt App

    Mul, M.F.; Becker, P.M.; Peet-Schwering, van der C.M.C.; Wikselaar, van P.G.; Wisselink, H.J.; Stockhofe, N.


    De Animal Sciences Group van Wageningen UR heeft in opdracht van biologische varkenshouders onderzocht of het mogelijk is om in plaats van antibiotica, knoflook te gebruiken voor de bestrijding van longontsteking door de bacterie Actinobacillus pleuropneumoniae (App). Uit de resultaten blijkt dat Al

  19. Susceptibility of bacteria isolated from pigs to tiamulin and enrofloxacin metabolites

    Lykkeberg, Anne Kruse; Halling-Sørensen, Bent; Jensen, Lars Bogø

    -tiamulin (8 alpha-HTIA), and the ENR metabolites were: ciprofloxacin (CIP) and enrofloxacin N-oxide (ENR-N). Bacteria, all of porcine origin, we're selected as representatives of bacterial infections (Stap4ylococcus hyicus and Actinobacillus pleuropneumoniae), zoonotic bacteria (Campylobacter coli) and...

  20. A Pair of Novel Primers for Universal Detection of the NS1 Gene from Various Bluetongue Virus Serotypes

    Hui-qiong YIN; Gai-ping ZHANG; Hong ZHANG; Jin-gang ZHANG


    Twenty five serotypes of Bluetongue virus (BTV) have been identified worldwide. Rapid and reliable methods of virus universal detection are essential for fighting against bluetongue (BT). We have therefore developed and evaluated a pair of primers which can detect various serotypes of BTV by RT-PCR. Analysis of the viral protein 7 (VP7) and the non-structural protein (NS1) gene from different serotypes of BTV by DNAstar showed that the 5' end of the NS1 gene is the most conserved region. The primer pairs (P1 and P2) were designed based on the highly conserved region of NS1. The novel primers were evaluated by detecting BTV serotypes 1, 3, 5, 8, 10, 11, 21 and 22. The specificity of the primers was estimated by comparing to gene sequences of viruses published in GenBank, and further assessed by detecting BTV serotype 1-12 and Epizootic hemorrhagic disease virus (EHDV) serotype 1-4. The sensitivity and repeatability of PCR with the novel primers were evaluated by successfully detecting the recombinant plasmid pGEM-T121 containing the diagnosed nucleotide sequence. Our results suggest that these unique primers can be used in high throughout and universal detection of the NS1 gene from various BTV serotypes.

  1. Serotypes, antibiotic susceptibilities, and multi-locus sequence type profiles of Streptococcus agalactiae isolates circulating in Beijing, China.

    Ping Wang

    Full Text Available To investigate the serotypes, antibiotic susceptibilities, and multi-locus sequence type (MLST profiles of Streptococcus agalactiae (S. agalactiae in Beijing to provide references for the prevention and treatment of S. agalactiae infections.All isolates were identified using the CAMP test and the latex-agglutination assay and serotyped using a Strep-B-Latex kit, after which they were assessed for antibiotic susceptibility, macrolide-resistance genes, and MLST profiles.In total, 56 S. agalactiae isolates were identified in 863 pregnant women (6.5%. Serotypes Ia, Ib, II, III, and V were identified, among which types III (32.1%, Ia (17.9%, Ib (16.1%, and V (14.3% were the predominant serotypes. All isolates were susceptible to penicillin and ceftriaxone. The nonsusceptiblity rates measured for erythromycin, clarithromycin, azithromycin, telithromycin, clindamycin, tetracycline, and levofloxacin were 85.7%, 92.9%, 98.2%, 30.4%, 73.2%, 91%, and 39.3%, respectively. We identified 14 sequence types (STs for the 56 isolates, among which ST19 (30.4% was predominant. The rate of fluoroquinolone resistance was higher in serotype III than in the other serotypes. Among the 44 erythromycin-resistant isolates, 32 (72.7% carried ermB.S. agalactiae isolates of the serotypes Ia, Ib, III, and V are common in Beijing. Among the S. agalactiae isolates, the macrolide and clindamycin resistance rates are extremely high. Most of the erythromycin-resistant isolates carry ermB.

  2. Concurrent infections by all four dengue virus serotypes during an outbreak of dengue in 2006 in Delhi, India

    Guleria Randeep


    Full Text Available Abstract Background Co-circulation of multiple dengue virus serotypes has been reported from many parts of the world including India, however concurrent infection with more than one serotype of dengue viruses in the same individual is rarely documented. An outbreak of dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS occurred in and around Delhi in 2006. This is the first report from India with high percentage of concurrent infections with different dengue virus serotypes circulating during one outbreak. Results Acute phase sera from patients were tested for the presence of dengue virus RNA by RT-PCR assay. Of the 69 samples tested for dengue virus RNA, 48 (69.5% were found to be positive. All the four dengue virus serotypes were found to be co-circulating in this outbreak with DENV-3 being the predominant serotype. In addition in 9 of 48 (19% dengue virus positive samples, concurrent infection with more than one dengue virus serotype were identified. Conclusion This is the first report in which concurrent infections with different dengue virus serotypes is being reported during an outbreak from India. Delhi is now truly hyperendemic for dengue.

  3. Rapid and Simultaneous Identification of Two Salmonella enterica Serotypes, Enteritidis and Typhimurium from Chicken and Meat Products by Multiplex PCR

    Hanan Issa Malkawi


    Full Text Available In order to overcome the large volume of work required to detect and identify food samples that may contain Salmonella enterica serotypes Enteritidis �and Typhimurium, multiplex polymerase chain reaction (m-PCR was used as a diagnostic tool to identify Salmonella enterica, Salmonella enterica ser. Enteritidis and Typhimurium in naturally contaminated meat and poultry products. Three sets of known S. enterica and serotypes S. enteritidis and S. typhimurium specific primers were applied to simultaneous identification of such pathogens in the most frequently used fresh and frozen meat (beef and lamb and poultry (chicken products (whole, cut, ground and processed collected from eight locations within Irbid city (Jordan. Out of 93 samples, Only 21 (22.5% samples contained serotype S. typhimurium alone. Nineteen samples (20% showed the 312 bp specific band for serotype S. enteritidis specific band. A total of 28 (30% samples showed only for S. enterica genus and 25 (26.8% samples showed both serotypes S. enteritidis and S. typhimurium. In general, 46 (49.5% samples contained serotype S. typhimurium, while 44 (47.3% samples contained serotype S. enteritidis. Multiplex PCR amplification was shown to be an effective and rapid method for the simultaneous identification of such pathogens in food samples.

  4. Geo-spatial distribution of serologically detected bovine Foot and Mouth Disease (FMD serotype outbreaks in Ilesha Baruba, Kwara State-Nigeria

    Hamza Olatunde Olabode


    Full Text Available The study was aimed at assessing the prevalence and distribution of bovine Foot and Mouth Disease (FMD serotypes in Ilesha Baruba, Kwara state-Nigeria. To identify the source of epidemics, geo-spatial analysis was done on the FMD outbreak locations (n=15 using Global Positioning Service (GPS device (EtrexR. Randomly sampled bovine sera (n=64 from herd representatives were subjected to FMD 3ABC enzyme-linked immunosorbent assay (FMD 3ABC ELISA and solid-phase competitive ELISA (SP-cELISA, for the screening and serotyping of FMD virus, respectively. Through ELISA, the FMD serotypes detected in this study were- serotype O (83%; n=53/64, serotype A (7.8%; n=5/64, serotype vaccine O (1.6%; n=1/64, and serotype vaccine SAT2 (1.6%; n=1/64. Multiple serotypes were observed in two different combinations; these were O and A (4.7%; n=3/64, and O and SAT2 (1.6%; n=1/64. FMD multiple serotype infections were associated with absence of cross-immunity between serotypes and cross reactivity enhanced by clustered herds, highland study area topography, road and river interconnectivity, possible human settlements, activities and traffic. This study provides baseline information on geo-spatial distribution, and identification of prevalent FMD serotypes in Ilesha Baruba, Kwara state-Nigeria.

  5. Re-emergence of dengue virus serotype 2 strains in the 2013 outbreak in Nepal

    Birendra Prasad Gupta


    Full Text Available Background & objectives: Epidemiological interventions and mosquito control are the available measures for dengue control. The former approach uses serotype and genetic information on the circulating virus strains. Dengue has been frequently reported from Nepal, but this information is mostly lacking. The present study was done to generate a comprehensive clinical and virological picture of a dengue outbreak in Nepal during 2013. Methods: A hospital-based study involving patients from five districts of Nepal was carried out. Demographic information, clinical details and dengue serological status were obtained. Viral RNA was characterized at the molecular level by reverse-transcription polymerase chain reaction (RT-PCR, nucleotide sequencing and phylogenetic analysis. Results: From among the 2340 laboratory-confirmed dengue cases during the study period, 198 patients consented for the study. Clinically they had fever (100%, headache (59.1%, rashes (18.2%, retro-orbital pain (30.3%, vomiting (15.1%, joint pain (28.8% and thrombocytopenia (74.3%. Fifteen (7.5% of them had mucosal bleeding manifestations, and the rest were uncomplicated dengue fever. The patients were mostly adults with a mean age of 45.75 ± 38.61 yr. Of the 52 acute serum samples tested, 15 were positive in RT-PCR. The causative virus was identified as DENV serotype 2 belonging to the Cosmopolitan genotype. Interpretations & conclusions: We report here the involvement of DENV serotype 2 in an outbreak in Nepal in 2013. Earlier outbreaks in the region in 2010 were attributed to serotype 1 virus. As serotype shifts are frequently associated with secondary infections and severe disease, there is a need for enhancing surveillance especially in the monsoon and post-monsoon periods to prevent large-scale, severe dengue outbreaks in the region.

  6. Clustering of serotypes in a longitudinal study of Streptococcus pneumoniae carriage in three day care centres

    Tanskanen Antti


    Full Text Available Abstract Background Streptococcus pneumoniae (pneumococcus causes a wide range of clinical manifestations that together constitute a major burden of disease worldwide. The main route of pneumococcal transmission is through asymptomatic colonisation of the nasopharynx. Studies of transmission are currently of general interest because of the impact of the new conjugate-polysaccharide vaccines on nasopharyngeal colonisation (carriage. Here we report the first longitudinal study of pneumococcal carriage that records serotype specific exposure to pneumococci simultaneously within the two most important mixing groups, families and day care facilities. Methods We followed attendees (N = 59 with their family members (N = 117 and the employees (N = 37 in three Finnish day care centres for 9 months with monthly sampling of nasopharyngeal carriage. Pneumococci were cultured, identified and serotyped by standard methods. Results Children in day care constitute a core group of pneumococcal carriage: of the 36 acquisitions of carriage with documented exposure to homologous pneumococci, the attendee had been exposed in her/his day care centre in 35 cases and in the family in 9 cases. Day care children introduce pneumococci to the family: 66% of acquisitions of a new serotype in a family were associated with simultaneous or previous carriage of the same type in the child attending day care. Consequently, pneumococcal transmission was found to take place as micro-epidemics driven by the day care centres. Each of the three day care centres was dominated by a serotype of its own, accounting for 100% of the isolates of that serotype among all samples from the day care attendees. Conclusion The transmission of pneumococci is more intense within than across clusters defined by day care facilities. The ensuing micro-epidemic behaviour enhances pneumococcal transmission.

  7. Clustering of serotypes in a longitudinal study of Streptococcus pneumoniae carriage in three day care centres

    Leino, Tuija; Hoti, Fabian; Syrjänen, Ritva; Tanskanen, Antti; Auranen, Kari


    Background Streptococcus pneumoniae (pneumococcus) causes a wide range of clinical manifestations that together constitute a major burden of disease worldwide. The main route of pneumococcal transmission is through asymptomatic colonisation of the nasopharynx. Studies of transmission are currently of general interest because of the impact of the new conjugate-polysaccharide vaccines on nasopharyngeal colonisation (carriage). Here we report the first longitudinal study of pneumococcal carriage that records serotype specific exposure to pneumococci simultaneously within the two most important mixing groups, families and day care facilities. Methods We followed attendees (N = 59) with their family members (N = 117) and the employees (N = 37) in three Finnish day care centres for 9 months with monthly sampling of nasopharyngeal carriage. Pneumococci were cultured, identified and serotyped by standard methods. Results Children in day care constitute a core group of pneumococcal carriage: of the 36 acquisitions of carriage with documented exposure to homologous pneumococci, the attendee had been exposed in her/his day care centre in 35 cases and in the family in 9 cases. Day care children introduce pneumococci to the family: 66% of acquisitions of a new serotype in a family were associated with simultaneous or previous carriage of the same type in the child attending day care. Consequently, pneumococcal transmission was found to take place as micro-epidemics driven by the day care centres. Each of the three day care centres was dominated by a serotype of its own, accounting for 100% of the isolates of that serotype among all samples from the day care attendees. Conclusion The transmission of pneumococci is more intense within than across clusters defined by day care facilities. The ensuing micro-epidemic behaviour enhances pneumococcal transmission. PMID:19116005

  8. Replication, neurotropism, and pathogenicity of avian paramyxovirus serotypes 1-9 in chickens and ducks.

    Shin-Hee Kim

    Full Text Available Avian paramyxovirus (APMV serotypes 1-9 have been isolated from many different avian species. APMV-1 (Newcastle disease virus is the only well-characterized serotype, because of the high morbidity, mortality, and economic loss caused by highly virulent strains. Very little is known about the pathogenesis, replication, virulence, and tropism of the other APMV serotypes. Here, this was evaluated for prototypes strains of APMV serotypes 2-9 in cell culture and in chickens and ducks. In cell culture, only APMV-1, -3 and -5 induced syncytium formation. In chicken DF1 cells, APMV-3 replicated with an efficiency approaching that of APMV-1, while APMV-2 and -5 replicated to lower, intermediate titers and the others were much lower. Mean death time (MDT assay in chicken eggs and intracerebral pathogenicity index (ICPI test in 1-day-old SPF chicks demonstrated that APMV types 2-9 were avirulent. Evaluation of replication in primary neuronal cells in vitro as well as in the brains of 1-day-old chicks showed that, among types 2-9, only APMV-3 was neurotropic, although this virus was not neurovirulent. Following intranasal infection of 1-day-old and 2-week-old chickens, replication of APMV types 2-9 was mostly restricted to the respiratory tract, although APMV-3 was neuroinvasive and neurotropic (but not neurovirulent and also was found in the spleen. Experimental intranasal infection of 3-week-old mallard ducks with the APMVs did not produce any clinical signs (even for APMV-1 and exhibited restricted viral replication of the APMVs (including APMV-1 to the upper respiratory tract regardless of their isolation source, indicating avirulence of APMV types 1-9 in mallard ducks. The link between the presence of a furin cleavage site in the F protein, syncytium formation, systemic spread, and virulence that has been well-established with APMV-1 pathotypes was not evident with the other APMV serotypes.

  9. European Bluetongue Serotype 8: Disease Threat Assessment for U.S. Sheep.

    Drolet, Barbara S; Reister-Hendricks, Lindsey M; Podell, Brendan K; Breitenbach, Jonathan E; McVey, D Scott; van Rijn, Piet A; Bowen, Richard A


    Bluetongue virus (BTV) is an orbivirus transmitted by biting midges (Culicoides spp.) that can result in moderate to high morbidity and mortality primarily in sheep and white-tailed deer. Although only 5 serotypes of BTV are considered endemic to the United States, as many as 11 incursive serotypes have been detected in livestock and wildlife in the past 16 years. Introductions of serotypes, with unknown virulence and disease risk, are constant threats to US agriculture. One potential incursive serotype of particular concern is the European strain of BTV-8, which was introduced into Northern Europe in 2006 and caused unprecedented livestock disease and mortality during the 2006-2007 vector seasons. To assess disease risk of BTV-8 in a common white-faced American sheep breed, eight Polled Dorset yearlings were experimentally infected and monitored for clinical signs. Viremia and viral tissue distribution were detected and quantified by real-time qRT-PCR. Overall, clinical disease was moderate with no mortality. Viremia reached as high as 9.7 log10 particles/mL and persisted at 5 logs or higher through the end of the study (28 days). Virus distribution in tissues was extensive with the highest mean titers at the peak of viremia (day 8) in the kidney (8.38 log10 particles/mg) and pancreas (8.37 log10 particles/mg). Virus persisted in tissues of some sheep at 8 logs or higher by day 28. Results of this study suggest that should BTV-8 emerge in the United States, clinical disease in this common sheep breed would likely be similar in form, duration, and severity to what is typically observed in severe outbreaks of endemic serotypes, not the extraordinary disease levels seen in Northern Europe. In addition, a majority of exposed sheep would be expected to survive and act as significant BTV-8 reservoirs with high titer viremias for subsequent transmission to other livestock and wildlife populations. PMID:27111674

  10. Experimental infection of hamsters with avian paramyxovirus serotypes 1 to 9

    Samuel Arthur S


    Full Text Available Abstract Avian paramyxoviruses (APMVs are frequently isolated from domestic and wild birds throughout the world and are separated into nine serotypes (APMV-1 to -9. Only in the case of APMV-1, the infection of non-avian species has been investigated. The APMVs presently are being considered as human vaccine vectors. In this study, we evaluated the replication and pathogenicity of all nine APMV serotypes in hamsters. The hamsters were inoculated intranasally with each virus and monitored for clinical disease, pathology, histopathology, virus replication, and seroconversion. On the basis of one or more of these criteria, each of the APMV serotypes was found to replicate in hamsters. The APMVs produced mild or inapparent clinical signs in hamsters except for APMV-9, which produced moderate disease. Gross lesions were observed over the pulmonary surface of hamsters infected with APMV-2 & -3, which showed petechial and ecchymotic hemorrhages, respectively. Replication of all of the APMVs except APMV-5 was confirmed in the nasal turbinates and lungs, indicating a tropism for the respiratory tract. Histologically, the infection resulted in lung lesions consistent with bronchointerstitial pneumonia of varying severity and nasal turbinates with blunting or loss of cilia of the epithelium lining the nasal septa. The majority of APMV-infected hamsters exhibited transient histological lesions that self resolved by 14 days post infection (dpi. All of the hamsters infected with the APMVs produced serotype-specific HI or neutralizing antibodies, confirming virus replication. Taken together, these results demonstrate that all nine known APMV serotypes are capable of replicating in hamsters with minimal disease and pathology.

  11. Multidrug resistance in Salmonella enterica serotype Typhimurium from humans in France (1993 to 2003).

    Weill, François-Xavier; Guesnier, Françoise; Guibert, Véronique; Timinouni, Mohammed; Demartin, Marie; Polomack, Lucette; Grimont, Patrick A D


    The aim of this study was to determine the distribution of the antimicrobial resistance phenotypes (R types), the phage types and XbaI-pulsed-field gel electrophoresis (PFGE) types, the genes coding for resistance to beta-lactams and to quinolones, and the class 1 integrons among a representative sample of Salmonella enterica serotype Typhimurium isolates collected from humans in 2002 through the French National Reference Center for Salmonella (NRC-Salm) network. The trends in the evolution of antimicrobial resistance of serotype Typhimurium were reviewed by using NRC-Salm data from 1993, 1997, 2000, and 2003. In 2002, 3,998 isolates of serotype Typhimurium were registered at the NRC-Salm among 11,775 serotyped S. enterica isolates (34%). The most common multiple antibiotic resistance pattern was resistance to amoxicillin, chloramphenicol, streptomycin and spectinomycin, sulfonamides, and tetracycline (ACSSpSuTe R type), with 156 isolates (48.8%). One isolate resistant to extended-spectrum cephalosporins due to the production of TEM-52 extended-spectrum beta-lactamase was detected (0.3%), and one multidrug-resistant isolate was highly resistant to ciprofloxacin (MIC > 32 mg/liter). We found that 57.2% of the isolates tested belonged to the DT104 clone. The main resistance pattern of DT104 isolates was R type ACSSpSuTe (83.2%). However, evolutionary changes have occurred within DT104, involving both loss (variants of Salmonella genomic island 1) and acquisition of genes for drug resistance to trimethoprim or to quinolones. PFGE profile X1 was the most prevalent (74.5%) among DT104 isolates, indicating the need to use a more discriminatory subtyping method for such isolates. Global data from the NRC-Salm suggested that DT104 was the main cause of multidrug resistance in serotype Typhimurium from humans from at least 1997 to 2003, with a roughly stable prevalence during this period. PMID:16517842

  12. Classification of Salmonella enterica serotypes with selective bands using visible/NIR hyperspectral microscope images.

    Eady, M; Park, B


    Optical detection of foodborne bacteria such as Salmonella classifies bacteria by analysing spectral data, and has potential for rapid detection. In this experiment hyperspectral microscopy is explored as a means for classifying five Salmonella serotypes. Initially, the microscope collects 89 spectral measurements between 450 and 800 nm. Here, the objective was to develop correct classification of five serotypes with optimal spectral bands selected through multivariate data analysis (MVDA), thus reducing the data processing and storage requirement necessary for practical application in the food industry. An upright digital microscope is equipped with an acousto-optical tuneable filter, electron multiplying charge-coupled device, and metal halide lighting source. Images for each of the five serotypes were collected, and informative bands were identified through a principal component analysis, for four abbreviated spectral ranges containing 3, 7, 12 and 20 spectral bands. The experiment was repeated with an independent repetition and images were collected at each of the reduced band sets, identified by the first repetition. A support vector machine (SVM) was used to classify serotypes. Results showed that with the first repetition, classification accuracy decreased from 99.5% (89 bands) to 84.5% (3 bands), whereas the second repetition showed classification accuracies of 100%, possibly due to a reduction in spectral noise. The support vector machine regression (SVMR) was applied with cross-validation, and had R(2) calibration and validation values >0.922. Although classification accuracies through SVM classification showed that as little as 3 bands were able to classify 100% of the samples, the SVMR shows that the smallest root-mean squared-error values were 0.001 and 0.002 for 20 and 12 bands, respectively, suggesting that the 12 band range collected between 586 and 630 nm is optimal for classifying bacterial serotypes, with only the informative HMI bands selected

  13. Cariogenicity of a lactate dehydrogenase-deficient mutant of Streptococcus mutans serotype c in gnotobiotic rats.

    Fitzgerald, R J; Adams, B O; Sandham, H J; Abhyankar, S


    A lactate dehydrogenase-deficient (Ldh-) mutant of a human isolate of Streptococcus mutans serotype c was tested in a gnotobiotic rat caries model. Compared with the wild-type Ldh-positive (Ldh+) strains, it was significantly (alpha less than or equal to 0.005) less cariogenic in experiments with two different sublines of Sprague-Dawley rats. The Ldh- mutant strain 044 colonized the oral cavity of the test animals to the same extent as its parent strain 041, although its initial implantation was slightly but not significantly (P greater than or equal to 0.2) less. Multiple oral or fecal samples plated on 2,3,5-triphenyltetrazolium indicator medium revealed no evidence of back mutation from Ldh- to Ldh+ in vivo. Both Ldh+ strain 041 and Ldh- strain 044 demonstrated bacteriocinlike activity in vitro against a number of human strains of mutans streptococci representing serotype a (S. cricetus) and serotypes c and e (S. mutans). Serotypes b (S. rattus) and f (S. mutans) and strains of S. mitior, S. sanguis, and S. salivarius were not inhibited. Thus, Ldh mutant strain 044 possesses a number of desirable traits that suggest it should be investigated further as a possible effector strain for replacement therapy of dental caries. These traits include its stability and low cariogenicity in the sensitive gnotobiotic rat caries model, its bacteriocinlike activity against certain other cariogenic S. mutans (but not against more inocuous indigenous oral streptococci), and the fact that it is a member of the most prevalent human serotype of cariogenic streptococci. PMID:2917788

  14. Association between infection of different strains of Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in subgingival plaque and clinical parameters in chronic periodontitis

    WU Yan-min; YAN Jie; CHEN Li-li; GU Zhi-yuan


    Objective: The aim of this study was to investigate subgingival infection frequencies ofPorphyromonas gtngivalis and Actinobacillus actinomycetemcomitans strains with genetic variation in Chinese chronic periodontitis (CP) patients and to evaluate its correlation with clinical parameters. Methods: Two multiplex polymerase chain reaction (PCR) assays were developed to detect the 16SrDNA, collagenase (prtC) and fimbria (fimA) genes of P. gingivalis and the 16SrDNA, leukotoxin (lktA) and fimbria-associated protein (lap) genes ofA. actinomycetemcomitans in 60 sulcus samples from 30 periodontal healthy subjects and in 122 subgingival plaque samples from 61 patients with CP. The PCR products were further T-A cloned and sent for nucleotide sequence analysis. Results: The 16SrDNA, prtC andfimA genes ofP. gingivalis were detected in 92.6%, 85.2% and 80.3% of the subgingival plaque samples respectively, while the 16SrDNA, lktA andfap genes ofA. actinomycetemcomitans were in 84.4%,75.4% and 50.0% respectively. Nucleotide sequence analysis showed 98.62%~100% homology of the PCR products in these genes with the reported sequences. P. gingivalis strains with prtC+/fimA+ and A. actinomycetemcomitans with lktA+ were predominant in deep pockets (>6 mm) or in sites with attachment loss ≥5 mm than in shallow pockets (3~4 mm) or in sites with attachment loss ≤2 mm (P<0.05). P. gingivalis strains with prtC+/fimA+ also showed higher frequency in gingival index (GI)=3than in GI= 1 group (P<0.05). Conclusion: Infection of P. gingivalis with prtC+/fimA+ and A. actinomycetemcomitans with lktA+correlates with periodontal destruction of CP in Chinese. Nonetheless P. gingivalis fimA, prtC genes and A. actinomycetemcomitans IktA gene are closely associated with periodontal destruction, while A. actinomycetemcomitansfap gene is not.

  15. Streptococcus pneumoniae Serotypes and Mortality in Adults and Adolescents in South Africa: Analysis of National Surveillance Data, 2003 - 2008.

    Cheryl Cohen

    Full Text Available An association between pneumococcal serotypes and mortality has been suggested. We aimed to investigate this among individuals aged ≥15 years with invasive pneumococcal disease (IPD in South Africa.IPD cases were identified through national laboratory-based surveillance at 25 sites, pre-pneumococcal conjugate vaccine (PCV introduction, from 2003-2008. We assessed the association between the 20 commonest serotypes and in-hospital mortality using logistic regression with serotype 4 (the third commonest serotype with intermediate case-fatality ratio (CFR as referent.Among 3953 IPD cases, CFR was 55% (641/1166 for meningitis and 23% (576/2484 for bacteremia (p<0.001. Serotype 19F had the highest CFR (48%, 100/207, followed by serotype 23F (39%, 99/252 and serotype 1 (38%, 246/651. On multivariable analysis, factors independently associated with mortality included serotype 1 (OR 1.9, 95%CI 1.1-3.5 and 19F (OR 2.9, 95%CI 1.4-6.1 vs. serotype 4; increasing age (25-44 years, OR 1.8, 95%CI 1.0-3.0; 45-64 years, OR 3.6, 95%CI 2.0-6.4; ≥65 years, OR 5.2, 95%CI 1.9-14.1; vs. 15-24 years; meningitis (OR 4.1, 95%CI 3.0-5.6 vs. bacteremic pneumonia; and HIV infection (OR1.7, 95%CI 1.0-2.8. On stratified multivariate analysis, serotype 19F was associated with increased mortality amongst bacteremic pneumococcal pneumonia cases, while no serotype was associated with increased mortality in meningitis cases.Mortality was increased in HIV-infected individuals, which may be reduced by increased antiretroviral therapy availability. Serotypes associated with increased mortality are included in the 10-and-13-valent PCV and may become less common in adults due to indirect effects following routine infant immunization.

  16. Serotype Distribution and Antimicrobial Resistance of Streptococcus pneumoniae prior to Introduction of the 10-Valent Pneumococcal Conjugate Vaccine in Brazil, 2000-2007: Pneumococcal Serotypes Prior to Conjugate Vaccine Introduction

    Menezes, Ana Paula de O.; Campos, Leila C.; dos Santos, Milena S.; Azevedo, Jailton; dos Santos, Renan Cardoso Nery; Carvalho, Maria da Gloria S.; Beall, Bernard W.; Martin, Stacey W.; Salgado, Katia; Reis, Mitermayer G.; Ko, Albert I.; Reis, Joice N


    This study describes the serotype distribution and antibiotic resistance patterns among 397 S. pneumoniae meningitis case isolates recovered in Salvador, Brazil, during the period of 2000-2007, before introduction of the 10-valent pneumococcal conjugate vaccine.

  17. Limited cross-reactivity of mouse monoclonal antibodies against Dengue virus capsid protein among four serotypes

    Noda M


    Full Text Available Megumi Noda,1 Promsin Masrinoul,1 Chaweewan Punkum,1 Chonlatip Pipattanaboon,2,3 Pongrama Ramasoota,2,4 Chayanee Setthapramote,2,3 Tadahiro Sasaki,6 Mikiko Sasayama,1 Akifumi Yamashita,1,5 Takeshi Kurosu,6 Kazuyoshi Ikuta,6 Tamaki Okabayashi11Mahidol-Osaka Center for Infectious Diseases, 2Center of Excellence for Antibody Research, 3Department of Microbiology and Immunology, 4Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Ratchathewi, Bangkok, Thailand; 5Graduate School of Life Science, Tohoku University, Sendai, Miyagi, 6Department of Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, JapanBackground: Dengue illness is one of the important mosquito-borne viral diseases in tropical and subtropical regions. Four serotypes of dengue virus (DENV-1, DENV-2, DENV-3, and DENV-4 are classified in the Flavivirus genus of the family Flaviviridae. We prepared monoclonal antibodies against DENV capsid protein from mice immunized with DENV-2 and determined the cross-reactivity with each serotype of DENV and Japanese encephalitis virus.Methods and results: To clarify the relationship between the cross-reactivity of monoclonal antibodies and the diversity of these viruses, we examined the situations of flaviviruses by analyses of phylogenetic trees. Among a total of 60 prepared monoclonal antibodies specific for DENV, five monoclonal antibodies stained the nuclei of infected cells and were found to be specific to the capsid protein. Three were specific to DENV-2, while the other two were cross-reactive with DENV-2 and DENV-4. No monoclonal antibodies were cross-reactive with all four serotypes. Phylogenetic analysis of DENV amino acid sequences of the capsid protein revealed that DENV-2 and DENV-4 were clustered in the same branch, while DENV-1 and DENV-3 were clustered in the other branch. However, these classifications of the capsid protein were different from those of the

  18. Comparação de métodos de isolamento de bactérias NAD-dependentes do trato respiratório superior de suínos sadios Methods to isolate NAD-dependent bacteria from the upper respiratory tract of health pigs

    J.D. Kich


    Full Text Available Secreções nasais, tonsilares e tecido tonsilar foram coletados de 67 leitões de 9 a 15 semanas de idade, provenientes de três rebanhos positivos para Actinobacillus pleuropneumoniae (App, e de 50 leitões provenientes de dois rebanhos negativos. Foram classificados como positivos aqueles rebanhos com isolamento prévio de sorotipos 3, 5 e 7 e rebanhos negativos aqueles submetidos a controle veterinário, sem notificação de sintomas clínicos, lesões de pleuropneumonia suína e sem isolamento do agente. O material coletado foi submetido a três diferentes métodos de cultivo: 1- semeadura direta em meio de cultivo sólido seletivo; 2- diluição em caldo seletivo seguido de subsemeadura em meio de cultivo sólido seletivo; 3- diluição em caldo seletivo seguido de subsemeadura em ágar sangue. Entre as amostras NAD-dependentes recuperadas 86 foram classificadas como App, 13 como grupo minor e 21 como grupo taxon (C, D, E e F. Dos rebanhos positivos foram recuperadas quatro amostras de App (sorotipos 3, 7 e 12 e 51 não sorotipificáveis. Dos rebanhos negativos foram recuperadas 31 amostras de App não sorotipificáveis, indicando que o App faz parte da flora normal do trato respiratório superior dos suínos. O melhor método de isolamento de amostras NAD-dependentes de leitões portadores foi da biópsia de tecido tonsilar semeado diretamente em meio sólido seletivo (PPLO ágar adicionado de 2mi g de cristal violeta, 10mi g NAD, 1mi g de lincomicina, 1,4mi g de bacitracina por ml.Nasal and tonsil secretions and tonsil tissue were collected from 67 piglets (9-to-15 weeks old from three positive herds and from 50 piglets from two negative herds. Positive herds were those in which an Actinobacillus pleuropneumoniae (App strain of serotype 3, 5 or 7 was isolated and negative ones those in which there was no notification of clinical symptoms or lesions of swine pleuropneumoniae, no App isolation and the herd was under veterinarian control

  19. Infants aged 12 months can mount adequate serotype-specific IgG responses to pneumococcal polysaccharide vaccine

    Balloch, Anne; Licciardi, Paul V.; Russell, Fiona M.; Edward K Mulholland; Tang, Mimi L. K.


    This is the first study examining serotype-specific IgG responses following immunization with the polysaccharide vaccine Pneumovax® in infants aged 12 months in the absence of prior pneumococcal conjugate vaccine priming.

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

    Stojiljković Jasmina


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

  1. Serotype- and virulence-associated gene profile of Streptococcus suis isolates from pig carcasses in Chiang Mai Province, Northern Thailand.

    Wongsawan, Kanruethai; Gottschalk, Marcelo; Tharavichitkul, Prasit


    In this present study, the serotype of 40 Streptococcus suis isolates from submaxillary glands of pig carcasses sold in wet markets in Chiang Mai Province, northern Thailand, was investigated. Eleven serotypes, including types 2, 3, 4, 5, 7, 8, 9, 17, 21, 22 and 31, were found in the isolates by a Multiplex PCR combined with serum agglutination. Of the eleven serotypes present, type 3 was the most prevalent, while types 2, 4, 5 and 21 were of primary interest due to their human isolate serotype. The mrp+/epf - /sly - genotype was found to be the most prevalent genotype. This study indicates the importance of effective control of human S. suis infection due to raw pork or pig carcass handling in northern Thailand. PMID:25367105

  2. Cloning and sequence analysis of hsf, an outer membrane protein gene of Pasteurella multocida serotype B:2

    A. Priyadarshini


    Full Text Available Aim: The present study was undertaken to clone, sequence and analyze the hsf, an outer membrane protein gene of Pasteurella multocida serotype B:2 Materials and Methods: hsf gene was amplified from genomic DNA of P. multocida. Polymerase chain reaction (PCR product was cloned in pET-32a vector and was characterized. hsf gene was sequenced, analyzed and phylogenetic tree was constructed taking sequences of other strains. Results: Amplicon size was found to be 785 bp. Recombinant got characterized through colony PCR and restriction enzyme analysis. Conclusion: hsf gene of P. multocida serotype B is similar to serotype A, but different from serotype D. Further work is needed to evaluate role of Hsf protein in protection studies and to study the antigenic properties of this recombinant protein as a candidate for vaccine.

  3. Rapid typing of foot-and-mouth disease serotype Asia 1 by reverse transcription loop-mediated isothermal amplification

    Chen Hao-tai


    Full Text Available Abstract A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP assay was rapidly used to detect serotype Asia 1 of foot-and-mouth disease virus (FMDV within 45 min at 61°C. All FMDV serotype Asia 1 reference strains were positive by RT-LAMP, while other viruses such as FMDV serotypes O, C, A and classical swine fever virus, swine vesicular disease virus, porcine reproductive and respiratory syndrome virus and Japanese encephalitis virus remained negative. Furthermore, FMDV sreotype Asia 1 positive samples were able to detect by RT-LAMP assay. This RT-LAMP assay may be suitable particularly for diagnosis of FMDV serotype Asia 1 infection in field stations.

  4. Genome sequences of serotype A6 Mannheimia haemolytica isolates D174 and D38 recovered from bovine pneumonia

    Here we report two genomes, one complete and one draft, from virulent bovine strains of Mannheimia haemolytica(strains D174 and D38)serotype A2 recovered prior to the field usage of modern antimicrobial drugs....

  5. Realtime PCR is more sensitive than multiplex PCR for diagnosis and serotyping in children with culture negative pneumococcal invasive disease.

    C. Azzari; M. Moriondo; G. INDOLFI; CORTIMIGLIA M; Canessa, C; BECCIOLINI L; F. Lippi; Martino, M; Resti, M


    Background Pneumococcal serotyping is usually performed by Quellung reaction, considered the gold standard test. However the method cannot be used on culture-negative samples. Molecular methods can be a useful alternative. The aim of the study was to evaluate the use of Multiplex-sequential-PCR (MS-PCR) or Realtime-PCR on blood samples for diagnosis and serotyping of invasive pneumococcal disease (IPD) in a pediatric clinical setting. Methodology/Principal Findings Sensitivity and specificity...

  6. Rates of Acquisition of Pneumococcal Colonization and Transmission Probabilities, by Serotype, Among Newborn Infants in Kilifi District, Kenya

    Tigoi, Caroline C.; Gatakaa, Hellen; Karani, Angela; Mugo, Daisy; Kungu, Stella; Wanjiru, Eva; Jomo, Jane; Musyimi, Robert; Ojal, John; Glass, Nina E.; Abdullahi, Osman; Scott, J Anthony G


    Background.  Herd protection and serotype replacement disease following introduction of pneumococcal conjugate vaccine (PCV) are attributable to the vaccine's impact on colonization. Prior to vaccine introduction in Kenya, we did an epidemiological study to estimate the rate of pneumococcal acquisition, by serotype, in an uncolonized population. Methods.  Nasopharyngeal swab specimens were taken from newborns aged ≤7 days and weekly thereafter for 13 weeks. Parents, and siblings aged

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

    Hofacre, Charles L.; Peter S. Holt; Lee, Margie D.; Susan Sanchez; Joseph Minicozzi; Maurer, John J.


    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 p...

  8. Genome sequence and comparative microarray analysis of serotype M18 group A Streptococcus strains associated with acute rheumatic fever outbreaks

    Smoot, James C; Barbian, Kent D.; Van Gompel, Jamie J.; Smoot, Laura M.; Chaussee, Michael S.; Sylva, Gail L.; Sturdevant, Daniel E.; Ricklefs, Stacy M.; Porcella, Stephen F.; Parkins, Larye D.; Beres, Stephen B.; Campbell, David S.; Smith, Todd M.; Zhang, Qing; Kapur, Vivek


    Acute rheumatic fever (ARF), a sequelae of group A Streptococcus (GAS) infection, is the most common cause of preventable childhood heart disease worldwide. The molecular basis of ARF and the subsequent rheumatic heart disease are poorly understood. Serotype M18 GAS strains have been associated for decades with ARF outbreaks in the U.S. As a first step toward gaining new insight into ARF pathogenesis, we sequenced the genome of strain MGAS8232, a serotype M18 organism isolated from a patient ...

  9. Validation of an Immunodiagnostic Assay for Detection of 13 Streptococcus pneumoniae Serotype-Specific Polysaccharides in Human Urine

    Pride, Michael W.; Huijts, Susanne M.; Wu, Kangjian; Souza, Victor; Passador, Sherry; Tinder, Chunyan; Song, Esther; Elfassy, Arik; McNeil, Lisa; Menton, Ronald; French, Roger; Callahan, Janice; Webber, Chris; Gruber, William C.; Bonten, Marc J. M.


    To improve the clinical diagnosis of pneumococcal infection in bacteremic and nonbacteremic community-acquired pneumonia (CAP), a Luminex technology-based multiplex urinary antigen detection (UAD) diagnostic assay was developed and validated. The UAD assay can simultaneously detect 13 different serotypes of Streptococcus pneumoniae by capturing serotype-specific S. pneumoniae polysaccharides (PnPSs) secreted in human urine. Assay specificity is achieved by capturing the polysaccharides with s...

  10. VP1 of serotype C foot-and-mouth disease viruses: long-term conservation of sequences.

    Piccone, M. E.; Kaplan, G; Giavedoni, L; Domingo, E; Palma, E L


    The nucleotide sequences of the VP1-coding regions of several isolates of serotype C3 foot-and-mouth disease virus (FMDV) were determined. The deduced amino acid sequences were compared with those of serotype C1 FMDV. The results provide evidence for two different lineages of FMDV C3 and document the potential for both long-term conservation and rapid evolution of FMDV.

  11. Dengue Virus Serotypes in Three Districts/Municipalities with Different Endemicity Level of Dengue in West Java

    Heni Prasetyowati


    Full Text Available The incidence rate of Dengue Hemorrhagic Fever (DHF disease in Indonesia is increasing over years. DHF outbreaks happen in many provinces of Indonesia. West Java is a DHF endemic province. Nearly all districts/municipalities at the West Java Province are endemic areas and have reported DHF outbreaks. Factors supporting high incidence rate of DHF are tropical climate of Indonesia and the circulation of four dengue virus serotypes. The study aimed to identify dengue virus serotype distribution in the districts with different DHF endemic at the Province of Jawa Barat.The study was observational with cross sectional design. Samples consisted of 60 samples of blood serum of patients serologically infected by dengue virus. Samples came from three districts/municipalities with different DHF endemic. Dengue virus serotype of samples was detected using nested RT-PCR (Reserve Transcription Polymerase Chain Reaction examination.Results showed that, four serotypes of dengue virus could be isolated from serum samples. Out of all positive samples, Den-2 was the serotype most frequently appeared (55% followed by Den-3 (29%, Den-1 (9.6% and Den-4 (6.4%. At dengue high endemic areas there were 4 serotypes of dengue virus Den-3 (6 times, Den-2(twice, Den-4 and Den-1 (once each. At medium endemic areas there were 4 serotypes of dengue virus, i.e. Den-2 (9 times, Den-3 (twice, Den-1 and Den-4 (once each. At low endemic areas there were two serotypes, i.e. Den-2 (6 times and Den-1 (once.

  12. Antigenic heterogeneity of capsid protein VP1 in foot-and-mouth disease virus (FMDV) serotype Asia 1

    Alam SM; Amin R; Rahman MZ; Hossain MA; Sultana M


    SM Sabbir Alam,1 Ruhul Amin,1 Mohammed Ziaur Rahman,2 M Anwar Hossain,1 Munawar Sultana11Department of Microbiology, University of Dhaka, Dhaka, Bangladesh; 2International Centre for Diarrhoeal Disease Research, Dhaka, BangladeshAbstract: Foot and mouth disease virus (FMDV), with its seven serotypes, is a highly contagious virus infecting mainly cloven-hoofed animals. The serotype Asia1 occurs mainly in Asian regions. An in-silico approach was taken to reveal the antigenic heterogeneities wit...

  13. Epidemiologic Typing of Salmonella enterica Serotype Enteritidis in a Canada-Wide Outbreak of Gastroenteritis due to Contaminated Cheese

    Ahmed, Rafiq; Soule, Geoff; Demczuk, Walter H.; Clark, Clifford; Khakhria, Rasik; Ratnam, Samuel; Marshall, Stephen; Ng, Lai-King; Woodward, David L.; Johnson, Wendy M; Rodgers, Frank G


    A major Canada-wide outbreak of gastroenteritis due to Salmonella enterica serotype Enteritidis phage type (PT) 8 occurred in 1998, and this was traced to contaminated cheese in a commercial lunch pack product. Phage typing and pulsed-field gel electrophoresis linked the clinical and cheese isolates of serotype Enteritidis but failed to differentiate outbreak from nonoutbreak PT 8 strains. Further differentiation was made by biotyping based on melibiose fermentation.

  14. Detection and serotyping of pneumococci in community acquired pneumonia patients without culture using blood and urine samples

    Elberse, K. (Karin); Mens, S.; Cremers, A.J.; Meijvis, S.C.A.; Vlaminckx, B.; de Jonge, M. I.; Meis, J. F. G. M.; Blauwendraat, C.; Pol, I. van de; Schouls, L. M.


    Background Treatment of community acquired pneumonia (CAP) patients with antibiotics before laboratory-confirmed diagnosis leads to loss of knowledge on the causative bacterial pathogen. Therefore, an increasing number of pneumococcal infections is identified using non-culture based techniques. However, methods for serotyping directly on the clinical specimen remain scarce. Here we present three approaches for detection and serotyping of pneumococci using samples from patients with CAP. Metho...

  15. The use of serotyping and PFGE-typing of Listeria monocytogenes in food processing contamination studies and human foodborne infections

    Sjöman, Maria


    Two different typing methods, serotyping and pulsed-field gel electrophoresis (PFGE) typing, were used to study Listeria monocytogenes contamination in food processing plants and human foodborne L. monocytogenes infections. L. monocytogenes contamination was studied in two different types of food processing plants: one ice cream plant and two broiler abattoirs. A dominant L. monocytogenes PFGE type of serotype 1/2b was found to have persisted in this ice cream plant for at least seven ...

  16. Direct and indirect interactions in the recognition between a cross-neutralizing antibody and the four serotypes of dengue virus.

    Lisova, Olesia; Belkadi, Laurent; Bedouelle, Hugues


    Dengue fever is the most important vector-borne viral disease. Four serotypes of dengue virus, DENV1 to DENV4, coexist. Secondary infection by a different serotype is a risk factor for severe dengue. Monoclonal antibody mAb4E11 neutralizes the four serotypes of DENV with varying efficacies by recognizing an epitope located within domain-III (ED3) of the viral envelope (E) protein. To better understand the cross-reactivities between mAb4E11 and the four serotypes of DENV, we constructed mutations in both Fab4E11 fragment and ED3, and we searched for indirect interactions in the crystal structures of the four complexes. According to the serotype, 7 to 12 interactions are mediated by one water molecule, 1 to 10 by two water molecules, and several of these interactions are conserved between serotypes. Most interfacial water molecules make hydrogen bonds with both antibody and antigen. Some residues or atomic groups are engaged in both direct and water-mediated interactions. The doubly-indirect interactions are more numerous in the complex of lowest affinity. The third complementarity determining region of the light chain (L-CDR3) of mAb4E11 does not contact ED3. The structures and double-mutant thermodynamic cycles showed that the effects of (hyper)-mutations in L-CDR3 on affinity were caused by conformational changes and indirect interactions with ED3 through other CDRs. Exchanges of residues between ED3 serotypes showed that their effects on affinity were context dependent. Thus, conformational changes, structural context, and indirect interactions should be included when studying cross-reactivity between antibodies and different serotypes of viral antigens for a better design of diagnostics, vaccine, and therapeutic tools against DENV and other Flaviviruses. PMID:24591178

  17. Recombinant Bivalent Vaccine against Foot-and-Mouth Disease Virus Serotype O/A Infection in Guinea Pig

    Jian-Zhong YI; Ming-Qiu LIU; Cai-Zhu ZHU; Qiang ZHANG; Zu-Tian SHENG; Qing-Yun DU; Wei-Yao YAN; Zhao-Xin ZHENG


    In this study, two DNA fragments encoding amino acid (141-160)-(21-40)-(141-160) of the VP 1 of FMDV (foot-and-mouth disease virus) serotype O and (138-160)-(21-40)-( 138-160) of the serotype A FMDV were chemically synthesized. These two tandem-repeat fragments were ligated and transfected into prokaryotic expression vector pTrcHis A to construct pTH-O-A. The other vector called pTH-O-scIgG-A was constructed similarly only that the two tandem-repeat DNA fragments were linked by the bovineIgG heavy chain coding sequence. Guinea pigs immunized with the two bivalent vaccines pTH-O-A and pTH-O-scIgG-A showed both specific antibody activity and T cell proliferation responses. FMDV challenge tests showed that 85% and 70% of guinea pigs vaccinated twice with 200 μg of the fusion protein of pTH-O-A were protected from FMDV serotype O and serotype A infection respectively. 70% and 57%of the guinea pigs immunized with the fusion protein of pTH-O-scIgG-A were protected from FMDV serotype O and serotype A infection respectively.

  18. Serotype and genotype diversity and hatchery transmission of Campylobacter jejuni in commercial poultry flocks

    Petersen, L.; Nielsen, E.M.; On, Stephen L.W.


    We investigated the genotype and serotype diversity of Campylobacter coli and C jejuni in two parent flocks of adult hens and their offspring over two rotations in order to evaluate the role of hatchery mediated transmission and/or vertical transmission of campylobacters in broiler flocks. In total......, 314 C jejuni and 32 C coli isolates from parent and broiler flocks and from the surroundings of broiler houses were typed by flagellin gene PCR/RFLP fla-typing), and selected isolates were also typed by serotyping and macrorestriction profiling using PFGE (MRP/PFGE). The combined typing results showed...... that the broiler flocks could be colonised by 1-3 different Campylobacter clones and parent flocks could be colonised by 2-6 different clones. C coli was isolated from up to 36% of birds in one parent flock, whereas only C jejuni was isolated from broiler flocks. C jejuni clones from different flocks...

  19. A Case of Necrotizing Fasciitis due to Streptococcus pneumoniae Serotype 5 in Saskatchewan

    Meenakshi Dawar


    Full Text Available Necrotizing fasciitis due to Streptococcus pneumoniae is a rare and grave condition, and only a few cases have been reported. Suggested risk factors include minor trauma, systemic lupus erythematosus, immunosuppression secondary to medication, use of intramuscular anti-inflammatories and alcoholism. A fatal case of pneumococcal necrotizing fasciitis that occurred in a 51-year-old woman with a history of alcohol abuse and oral anti-inflammatory use is presented. Her condition was caused by a multi-etiology outbreak of community-acquired pneumonia, from which S pneumoniae serotype 5 was also isolated. The case description outlines the subtle presentation and rapid clinical progression of this condition. Because serotype 5 antigen is included in the polysaccharide 23-valent pneumococcal vaccine, the present case highlights the importance of pneumococcal immunization programs in Canada.

  20. Characterization of the 2013 dengue epidemic in Myanmar with dengue virus 1 as the dominant serotype.

    Ngwe Tun, Mya Myat; Kyaw, Aung Kyaw; Makki, Nader; Muthugala, Rohitha; Nabeshima, Takeshi; Inoue, Shingo; Hayasaka, Daisuke; Moi, Meng Ling; Buerano, Corazon C; Thwe, Saw Myat; Thant, Kyaw Zin; Morita, Kouichi


    In 2013 in Myanmar, dengue epidemic occurred with 20,255 cases including 84 deaths. This study aimed to determine the serological and molecular characteristics of dengue virus (DENV) infection among children with clinical diagnosis of dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS) during this period. Single acute serum samples were collected from 300 children in Mandalay Children Hospital, Mandalay, Myanmar. Out of the 300 children, 175 (58.3%) and 183 (61%) were positive for anti-dengue IgM and anti-dengue IgG, respectively. Among the IgM positives, 41 (23.4%) had primary DENV infection. Thirty-nine DENV strains (23 DENV-1, 10 DENV-2 and 6 DENV-4) were successfully isolated after inoculation of the patient serum samples onto C6/36 cells. DENV 1 was the dominant serotype in the 2013 epidemic. There was no correlation between the infecting serotypes and clinical severities. The DENV-1 strains belonged to three lineages of the genotype 1; the DENV-2 strains were of the Asian I genotype and were separated into two lineages; and DENV-4 strains belonged to the same lineage of genotype I. It is of interest to note the diversity of DENV-1 and -2 circulating in the same location during June-August 2013. These DENV isolates were genetically close (98%-100%) to the other previously reported isolates from Myanmar and its neighboring countries, namely China, Thailand, Sri Lanka, Cambodia and Vietnam. Primary DENV infection was still high among the severe dengue cases. Different serotypes of DENV were co-circulating in 2013, however, genotype shift was not observed. Additionally, amino acid mutations were detected in the study strains not seen in the previously reported strains from other countries and Myanmar. This paper provided information on the circulating serotypes for the last 15years and the recent dengue situation in Mandalay, Myanmar after 2006. PMID:27154331

  1. Structural analysis of the receptor binding domain of botulinum neurotoxin serotype D

    Zhang, Yanfeng; Buchko, Garry W.; Qin, Lin; Robinson, Howard; Varnum, Susan M.


    Botulinum neurotoxins (BoNTs) are the most toxic proteins known. The mechanism for entry into neuronal cells for serotypes A, B, E, F, and G involves a well understood dual receptor (protein and ganglioside) process, however, the mechanism of entry for serotypes C and D remains unclear. To provide structural insights into how BoNT/D enters neuronal cells, the crystal structure of the receptor binding domain (S863-E1276) for this serotype (BoNT/D-HCR) was determined at 1.65 Å resolution. While BoNT/D-HCR adopts an overall fold similar to that observed in other known BoNT HCRs, several major structural differences are present. These structural differences are located at, or near, putative receptor binding sites and may be responsible for BoNT/D host preferences. Two loops, S1195-I1204 and K1236-N1244, located on both sides of the putative protein receptor binding pocket, are displaced >10 Å relative to the corresponding residues in the crystal structures of BoNT/B and G. Obvious clashes were observed in the putative protein receptor binding site when the BoNT/B protein receptor synaptotagmin II was modeled into the BoNT/D-HCR structure. Although a ganglioside binding site has never been unambiguously identified in BoNT/D-HCR, a shallow cavity in an analogous location to the other BoNT serotypes HCR domains is observed in BoNT/D-HCR that has features compatible with membrane binding. A portion of a loop near the putative receptor binding site, K1236-N1244, is hydrophobic and solvent-exposed and may directly bind membrane lipids. Liposome-binding experiments with BoNT/D-HCR demonstrate that this membrane lipid may be phosphatidylethanolamine.

  2. Novel Ganglioside-mediated Entry of Botulinum Neurotoxin Serotype D into Neurons

    Kroken, Abby R.; Karalewitz, Andrew P.-A.; Fu, Zhuji; Kim, Jung-Ja P.; Barbieri, Joseph T. (MCW)


    Botulinum Neurotoxins (BoNTs) are organized into seven serotypes, A-G. Although several BoNT serotypes enter neurons through synaptic vesicle cycling utilizing dual receptors (a ganglioside and a synaptic vesicle-associated protein), the entry pathway of BoNT/D is less well understood. Although BoNT/D entry is ganglioside-dependent, alignment and structural studies show that BoNT/D lacks key residues within a conserved ganglioside binding pocket that are present in BoNT serotypes A, B, E, F, and G, which indicate that BoNT/D-ganglioside interactions may be unique. In this study BoNT/D is shown to have a unique association with ganglioside relative to the other BoNT serotypes, utilizing a ganglioside binding loop (GBL, residues Tyr-1235-Ala-1245) within the receptor binding domain of BoNT/D (HCR/D) via b-series gangliosides, including GT1b, GD1b, and GD2. HCR/D bound gangliosides and entered neurons dependent upon the aromatic ring of Phe-1240 within the GBL. This is the first BoNT-ganglioside interaction that is mediated by a phenylalanine. In contrast, Trp-1238, located near the N terminus of the ganglioside binding loop, was mostly solvent-inaccessible and appeared to contribute to maintaining the loop structure. BoNT/D entry and intoxication were enhanced by membrane depolarization via synaptic vesicle cycling, where HCR/D colocalized with synaptophysin, a synaptic vesicle marker, but immunoprecipitation experiments did not detect direct association with synaptic vesicle protein 2. Thus, BoNT/D utilizes unique associations with gangliosides and synaptic vesicles to enter neurons, which may facilitate new neurotoxin therapies.

  3. Structural Analysis of the Receptor Binding Domain of Botulinum Neurotoxin Serotype D

    Y Zhang; G Buchko; L Qin; H Robinson; S Varnum


    Botulinum neurotoxins (BoNTs) are the most toxic proteins known. The mechanism for entry into neuronal cells for serotypes A, B, E, F, and G involves a well understood dual receptor (protein and ganglioside) process, however, the mechanism of entry for serotypes C and D remains unclear. To provide structural insights into how BoNT/D enters neuronal cells, the crystal structure of the receptor binding domain (S863-E1276) for this serotype (BoNT/D-HCR) was determined at 1.65{angstrom} resolution. While BoNT/D-HCR adopts an overall fold similar to that observed in other known BoNT HCRs, several major structural differences are present. These structural differences are located at, or near, putative receptor binding sites and may be responsible for BoNT/D host preferences. Two loops, S1195-I1204 and K1236-N1244, located on both sides of the putative protein receptor binding pocket, are displaced >10{angstrom} relative to the corresponding residues in the crystal structures of BoNT/B and G. Obvious clashes were observed in the putative protein receptor binding site when the BoNT/B protein receptor synaptotagmin II was modeled into the BoNT/D-HCR structure. Although a ganglioside binding site has never been unambiguously identified in BoNT/D-HCR, a shallow cavity in an analogous location to the other BoNT serotypes HCR domains is observed in BoNT/D-HCR that has features compatible with membrane binding. A portion of a loop near the putative receptor binding site, K1236-N1244, is hydrophobic and solvent-exposed and may directly bind membrane lipids. Liposome-binding experiments with BoNT/D-HCR demonstrate that this membrane lipid may be phosphatidylethanolamine.

  4. Culicoides fauna and bluetongue virus serotype 8 infection in South American camelid herds in Germany

    Schulz, Claudia


    Bluetongue (BT) is a Culicoides-born infectious disease caused by bluetongue virus (BTV). From 2006 to 2010, BTV serotype 8 (BTV-8) spread throughout Europe, causing severe disease in domestic and some wild ruminant species and in an alpaca. Compulsory vaccination of susceptible animals was the most effective strategy to control and eradicate the BTV-8 epizootic in Europe. However, South American camelids (SAC) were not included in the BTV-8 vaccination programmes in Europe. The presented...

  5. Immunity of Foot-and-Mouth Disease Serotype Asia 1 by Sublingual Vaccination

    Hao-tai Chen; Yong-sheng Liu


    Foot-and-mouth disease virus (FMDV) causes vesicular disease of cloven-hoofed animals, with severe agricultural and economic losses. Here we present study using a sublingual (SL) route with the killed serotype Asia 1 FMDV vaccine. Guinea pigs were vaccinated using a commercially available vaccine formulation at the manufacturer's recommended full, 1/4, and 1/16 antigen doses. Animals were challenged with homologous FMDV Asia1 strain at various times following vaccination. All control guinea p...

  6. Development of a Multiplex PCR Assay for Rapid Molecular Serotyping of Haemophilus parasuis

    Howell, Kate J; Peters, Sarah E; Wang, Jinhong; Hernandez-Garcia, Juan; Weinert, Lucy A; Luan, Shi-Lu; Chaudhuri, Roy R.; Angen, ?ystein; Aragon, Virginia; Williamson, Susanna M.; Parkhill, Julian; Langford, Paul R.; Rycroft, Andrew N.; Wren, Brendan W.; Maskell, Duncan J


    Haemophilus parasuis causes Glässer's disease and pneumonia in pigs. Indirect hemagglutination (IHA) is typically used to serotype this bacterium, distinguishing 15 serovars with some nontypeable isolates. The capsule loci of the 15 reference strains have been annotated, and significant genetic variation was identified between serovars, with the exception of serovars 5 and 12. A capsule locus and in silico serovar were identified for all but two nontypeable isolates in our collection of >200 ...

  7. Genome-Wide Association Studies of Virulent and Avirulent Haemophilus parasuis Serotype 4 Strains

    Lawrence, Paulraj K; Wiener, Brittanny L.; Kolander-Bremer, Tammy; Bey, Russell F.; Stine, Douglas L.; Kittichotirat, Weerayuth; Bumgarner, Roger E.


    Haemophilus parasuis is a normal commensal of the upper respiratory tract of healthy pigs. However, in conjunction with stress and/or viral infections, or in immunocompromised animals, H. parasuis can transform into a pathogen causing Glasser’s disease, which is typically characterized by fibrinous polyserositis, polyarthritis, meningitis, and sometimes acute pneumonia and septicemia. H. parasuis serotype 5 is highly virulent and more frequently isolated from respiratory and systemic infectio...

  8. Serotype Distribution, Antibiotic Resistance and Clonality of Streptococcus pneumoniae Isolated from Immunocompromised Patients in Tunisia

    Raddaoui, Anis; Simões, Alexandra S.; Baaboura, Rekaya; Félix, Sofia; Achour, Wafa; Ben Othman, Tarek; Béjaoui, Mohamed; Sá-Leão, Raquel; Ben Hassen, Assia


    Background Pneumococcal disease, a major cause of morbidity and mortality globally, has higher incidence among young children, the elderly and the immunocompromised of all ages. In Tunisia, pneumococcal conjugate vaccines (PCVs) are not included in the national immunization program. Also, few studies have described the epidemiology of S. pneumoniae in this country and, in particular, no molecular typing studies have been performed. The aim of this study was to evaluate serotype distribution, ...

  9. Epidemiology of serotype 19A isolates from invasive pneumococcal disease in German children

    van der Linden Mark


    Full Text Available Abstract Background This study presents an analysis of 159 serotype 19A isolates from IPD in children before and after the general recommendation for childhood pneumococcal conjugate vaccination in Germany in July 2006. Vaccination formulations used were PCV7, PCV10 (from April 2009 and PCV13 (from Dec. 2009, replacing PCV7. Methods Isolates from invasive pneumococcal disease in children were serotyped using the Quellung reaction, tested for antibiotic susceptibility and analysed for their multi locus sequence type. Results In an analysis of 3328 isolates from invasive pneumococcal disease (IPD in children that were sent to the German National Reference Center for Streptococci between July 1997 and June 2011, we show that the proportion of 19A isolates ranged between 1.7 and 4.2% in the period 1997 to 2006. After the recommendation for pneumococcal conjugate childhood vaccination, which was issued in July 2006, the proportion of 19A isolates increased significantly to 15.0% in 2010/11. Eight clonal complexes (CC and groups accounted for 77.2% and 65.3% of all serotype 19A isolates before and after vaccination, respectively. While three CCs and several STs were not detected after vaccine introduction, four CCs and several STs first appeared after vaccination, including three ST320 isolates that could be traced to recent imports from the US, UK and India. The proportion of penicillin-nonsusceptible and of multidrug-resistant 19A isolates moderately increased after vaccine introduction. A significant increase in the use of cephalosporins and azithromycin was noted post-vaccination (p=0.00001 and p=0.0013 respectively. Conclusions The prevalence of serotype 19A in Germany has increased significantly between July 2007 and June 2011. Possible reasons for this are the introduction of pneumococcal conjugate vaccination, increased use of cephalosporins and azithromycin, import of multidrug-resistant isolates and increased reporting.


    Jafari, A. A.


    After using 3 different generations of antibodies including human and non-human hyperimmune sera, monoclonal antibodies and chimeric antibodies, more recently a newer approach has been developed in which the antibody genes are cloned directly from a patient peripheral B-lymphocytes and expressed in a host like E. coli. In this study the Candida albicans serotype A (NCTC 3153) mannan was purified using a modified Fehling method and used for selection of human recombinant antibody from a C. alb...

  11. [Acute gastrointestinal involvement in dengue disease by serotype 4: a case report and literature review].

    Marín, Johan; Vilcarromero, Stalin; Forshey, Brett M; Celis-Salinas, Juan C; Ramal-Asayag, Cesar; Morrison, Amy C; Laguna-Torres, Alberto; Casapía, Martín; Halsey, Eric S


    Dengue fever is the world's most important arboviral disease, presenting a wide clinical spectrum. We report for the first time in Peru, a case caused by dengue virus serotype 4 with significant gastrointestinal involvement (acute acalculous cholecystitis and acute hepatitis). In addition we carried out a review of the literature atypical presentation illustrating the importance of the characteristics of abdominal pain (right upper quadrant); presence of Murphy's sign, ultrasound, and liver enzymes levels, for appropriate diagnosis and clinical management. PMID:24248170

  12. Capsular Gene Typing of Streptococcus agalactiae Compared to Serotyping by Latex Agglutination

    Yao, K.; Poulsen, K.; Maione, D.; Rinaudo, C. D.; Baldassarri, L.; Telford, J L; Sorensen, U. B. S.; Kilian, M.


    We evaluated three different PCR-based capsular gene typing methods applied to 312 human and bovine Streptococcus agalactiae (group B Streptococcus [GBS]) isolates and compared the results to serotyping results obtained by latex agglutination. Among 281 human isolates 27% could not be typed by latex agglutination. All 312 isolates except 5 could be typed by the three PCR methods combined. Two of these methods were multiplex assays. Among the isolates that were typeable by both latex agglutina...

  13. Complete Genome Sequence of Highly Virulent Haemophilus parasuis Serotype 11 Strain SC1401

    Dai, Ke; Jin, Jin; Wen, Xintian; He, Lvqin; Cao, Sanjie; Huang, Xiaobo; Wu, Rui; Zhao, Qin


    Haemophilus parasuis, a normal Gram-negative bacterium, may cause Glässer’s disease and pneumonia in pigs. This study aims to identify the genes related to natural competence of the serotype 11 strain SC1401, which frequently shows competence and high pathogenicity. SC1401 shows many differences from strains without natural competence within the molecular basis. We performed complete genome sequencing together with restriction modification system analysis to lay the foundation for later study. PMID:27445368

  14. PCR-based characterization of Yersinia enterocolitica : comparison with biotyping and serotyping

    Odinot, P. T.; Meis, J. F. G. M.; Hurk, P.J.J.C. van den; Hoogkamp-Korstanje, J. A. A.; Melchers, W. J. G.


    PCR-based DNA fingerprinting was used to characterize 48 clinical isolates of Yersinia enterocolitica. The samples were examined by random amplified polymorphic DNA (RAPD-PCR) and inter-repeat PCR (IR-PCR). IR-PCR with two enterobacterial repetitive intergenic consensus primers resulted in patterns which were poorly discriminated; 2 of 11 arbitrary primers (RAPD-PCR) provided sufficient discriminatory power. In comparisons with serotyping and biotyping, RAPD-fingerprinting was the most discri...

  15. Adeno-Associated Virus Serotype-9 Microdystrophin Gene Therapy Ameliorates Electrocardiographic Abnormalities in mdx Mice

    Bostick, Brian; Yue, Yongping; Lai, Yi; Long, Chun; Li, Dejia; Duan, Dongsheng


    Adeno-associated virus (AAV)-mediated microdystrophin gene therapy holds great promise for treating Duchenne muscular dystrophy (DMD). Previous studies have revealed excellent skeletal muscle protection. Cardiac muscle is also compromised in DMD patients. Here we show that a single intravenous injection of AAV serotype-9 (AAV-9) microdystrophin vector efficiently transduced the entire heart in neonatal mdx mice, a dystrophin-deficient mouse DMD model. Furthermore, microdystrophin therapy norm...

  16. Streptococcus pyogenes Serotype M1 Encodes Multiple Pathways for Entry into Human Epithelial Cells

    Cue, D; Dombek, P. E.; Lam, H; Cleary, P P


    The ability of a serotype M1 strain of Streptococcus pyogenes to efficiently invade A549 human lung epithelial cells was previously shown to be dependent on bacterial exposure to human or bovine serum proteins or synthetic peptides containing the sequence RGD. In this study, stimulation by invasion agonists was determined to be dependent on expression of the streptococcal cell surface protein, M1. Fetal bovine serum (FBS), fibronectin (Fn), the extracellular matrix protein laminin (Lm), and R...

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

    Mark Achtman

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

  18. Enhanced detection and serotyping of Streptococcus pneumoniae using multiplex polymerase chain reaction

    Jong Gyun Ahn


    Full Text Available &lt;B&gt;Purpose:&lt;/B&gt; Methods for quick and reliable detection of &lt;I&gt;Streptococcus pneumoniae&lt;/I&gt; are needed for the diagnosis of pneumococcal disease and vaccine studies. This study aimed to show that sequential multiplex polymerase chain reaction (PCR is more efficient than conventional culture in achieving &lt;I&gt;S. pneumoniae -positive&lt;/i&gt; results. &lt;B&gt;Methods:&lt;/B&gt; Nasopharyngeal (NP secretions were obtained from 842 pediatric patients admitted with lower respiratory infections at Severance Children’s Hospital in Korea between March 2009 and June 2010. For identification and serotype determination of pneumococci from the NP secretions, the secretions were evaluated via multiplex PCR technique with 35 serotype-specific primers arranged in 8 multiplex PCR sets and conventional bacteriological culture technique. &lt;B&gt;Results:&lt;/B&gt; Among the results for 793 samples that underwent both bacterial culture and PCR analysis for pneumococcal detection, 153 (19.3% results obtained by PCR and 81 (10.2% results obtained by conventional culture technique were positive for S. pneumoniae. The predominant serotypes observed, in order of decreasing frequency, were 19A (23%, 6A/B (16%, 19F (11%, 15B/C (5%, 15A (5%, and 11A (4%; further, 26% of the isolates were non-typeable. &lt;B&gt;Conclusion:&lt;/B&gt; As opposed to conventional bacteriological tests, PCR analysis can accurately and rapidly identify pneumococcal serotypes.

  19. Dengue Virus Serotype 2 from a Sylvatic Lineage Isolated from a Patient with Dengue Hemorrhagic Fever

    Jane Cardosa; Mong How Ooi; Phaik Hooi Tio; David Perera; Edward C Holmes; Khatijar Bibi; Zahara Abdul Manap


    Author Summary Dengue viruses are mosquito-borne RNA viruses that cause a spectrum of illness from mild disease to life-threatening dengue hemorrhagic fever (DHF). Dengue viruses exist in two separate cycles in nature, circulating in either non-human primates or humans. The viruses that are endemic in humans today most likely evolved from non-human primate dengue viruses a few hundred years ago and have since established themselves as four distinct serotypes in human populations, causing peri...

  20. Genome Sequence of Foot-and-Mouth Disease Virus Serotype O Isolated from Morocco in 2015.

    Bachanek-Bankowska, K; Wadsworth, J; Gray, A; Abouchoaib, N; King, D P; Knowles, N J


    The genome of a virus isolated from an outbreak of foot-and-mouth disease (FMD) in Morocco in 2015 is described here. This virus is classified as lineage Ind-2001d within serotype O, topotype ME-SA (Middle East-South Asia). This lineage is endemic on the Indian subcontinent but has caused outbreaks in the Middle East and North Africa since 2013. PMID:27103736

  1. Genome Sequence of Foot-and-Mouth Disease Virus Serotype O Isolated from Morocco in 2015

    Bachanek-Bankowska, K.; Wadsworth, J; Gray, A; Abouchoaib, N.; King, D.P.; Knowles, N.J.


    The genome of a virus isolated from an outbreak of foot-and-mouth disease (FMD) in Morocco in 2015 is described here. This virus is classified as lineage Ind-2001d within serotype O, topotype ME-SA (Middle East-South Asia). This lineage is endemic on the Indian subcontinent but has caused outbreaks in the Middle East and North Africa since 2013.

  2. All foot and mouth disease virus serotypes initiate protein synthesis at two separate AUGs.

    Sangar, D V; Newton, S E; Rowlands, D J; Clarke, B E


    Translation of the foot and mouth disease virus genome in vitro and in vivo indicated that all seven serotypes initiate protein synthesis at two separate AUGs. Sequence analysis of the region surrounding these AUGs has shown that the efficiency with which the initiating AUG is recognized is dependent on the flanking nucleotides. However, in vitro, the major factor determining which AUG is used is the concentration of Mg2+.

  3. Chromosomal Rearrangements in Salmonella enterica Serotype Typhi Affecting Molecular Typing in Outbreak Investigations

    Echeita, M. A.; Usera, M A


    Salmonella enterica serotype Typhi strains belonging to eight different outbreaks of typhoid fever that occurred in Spain between 1989 and 1994 were analyzed by ribotyping and pulsed-field gel electrophoresis. For three outbreaks, two different patterns were detected for each outbreak. The partial digestion analysis by the intron-encoded endonuclease I-CeuI of the two different strains from each outbreak provided an excellent tool for examining the organization of the genomes of epidemiologic...

  4. DNA microarray analysis of Salmonella serotype Typhimurium strains causing different symptoms of disease

    Helms Morten


    Full Text Available Abstract Background Salmonella enterica subsp. enterica is one of the leading food-borne pathogens in the USA and European countries. Outcome of human Salmonella serotype Typhimurium infections ranges from mild self-limiting diarrhoea to severe diarrhoea that requires hospitalization. Increased knowledge of the mechanisms that are responsible for causing infection and especially the severity of infection is of high interest. Results Strains were selected from patients with mild infections (n = 9 and patients with severe infections (n = 9 and clinical data allowed us to correct for known underlying diseases. Additionally, outbreak isolates (n = 3 were selected. Strains were analyzed on a DNA-DNA microarray for presence or absence of 281 genes covering marker groups of genes related to pathogenicity, phages, antimicrobial resistance, fimbriae, mobility, serotype and metabolism. Strains showed highly similar profiles when comparing virulence associated genes, but differences between strains were detected in the prophage marker group. The Salmonella virulence plasmid was present in 72% of the strains, but presence or absence of the virulence plasmid did not correspond to disease symptoms. A dendrogram clustered strains into four groups. Clustering confirmed DT104 as being a clonal phagetype. Clustering of the remaining strains was mainly correlated to presence or absence of the virulence plasmid and mobile elements such as transposons. Each of the four clusters in the tree represented an almost equal amount of strains causing severe or mild symptoms of infection. Conclusions We investigated clinical significance of known virulence factors of Salmonella serotype Typhimurium strains causing different disease symptoms, and conclude that the few detected differences in Salmonella serotype Typhimurium do not affect outcome of human disease.

  5. Towards the development of a DNA-sequence based approach to serotyping of Salmonella enterica

    Logan Julie MJ


    Full Text Available Abstract Background The fliC and fljB genes in Salmonella code for the phase 1 (H1 and phase 2 (H2 flagellin respectively, the rfb cluster encodes the majority of enzymes for polysaccharide (O antigen biosynthesis, together they determine the antigenic profile by which Salmonella are identified. Sequencing and characterisation of fliC was performed in the development of a molecular serotyping technique. Results FliC sequencing of 106 strains revealed two groups; the g-complex included those exhibiting "g" or "m,t" antigenic factors, and the non-g strains which formed a second more diverse group. Variation in fliC was characterised and sero-specific motifs identified. Furthermore, it was possible to identify differences in certain H antigens that are not detected by traditional serotyping. A rapid short sequencing assay was developed to target serotype-specific sequence motifs in fliC. The assay was evaluated for identification of H1 antigens with a panel of 55 strains. Conclusion FliC sequences were obtained for more than 100 strains comprising 29 different H1 alleles. Unique pyrosequencing profiles corresponding to the H1 component of the serotype were generated reproducibly for the 23 alleles represented in the evaluation panel. Short read sequence assays can now be used to identify fliC alleles in approximately 97% of the 50 medically most important Salmonella in England and Wales. Capability for high throughput testing and automation give these assays considerable advantages over traditional methods.

  6. Immunity of Foot-and-Mouth Disease Serotype Asia 1 by Sublingual Vaccination

    Chen, Hao-tai; Liu, Yong-sheng


    Foot-and-mouth disease virus (FMDV) causes vesicular disease of cloven-hoofed animals, with severe agricultural and economic losses. Here we present study using a sublingual (SL) route with the killed serotype Asia 1 FMDV vaccine. Guinea pigs were vaccinated using a commercially available vaccine formulation at the manufacturer’s recommended full, 1/4, and 1/16 antigen doses. Animals were challenged with homologous FMDV Asia1 strain at various times following vaccination. All control guinea p...

  7. Evaluation of different adjuvants for foot-and-mouth disease vaccine containing all the SAT serotypes

    Cloete, M.; B. Dungu; L.I. Van Staden; N. Ismail-Cassim; W. Vosloo


    Foot-and-mouth disease (FMD) is an economically important disease of cloven-hoofed animals that is primarily controlled by vaccination of susceptible animals and movement restrictions for animals and animal-derived products in South Africa. Vaccination using aluminium hydroxide gel-saponin (AS) adjuvanted vaccines containing the South African Territories (SAT) serotypes has been shown to be effective both in ensuring that disease does not spread from the endemic to the free zone and in ...

  8. Antimicrobial Resistance of Shigella flexneri Serotype 1b Isolates in China.

    Xianyan Cui

    Full Text Available Shigella flexneri serotype 1b is among the most prominent serotypes in developing countries, followed by serotype 2a. However, only limited data is available on the global phenotypic and genotypic characteristics of S. flexneri 1b. In the present study, 40 S. flexneri 1b isolates from different regions of China were confirmed by serotyping and biochemical characterization. Antimicrobial susceptibility testing showed that 85% of these isolates were multidrug-resistant strains and antibiotic susceptibility profiles varied between geographical locations. Strains from Yunnan were far more resistant than those from Xinjiang, while only one strain from Shanghai was resistant to ceftazidime and aztreonam. Fifteen cephalosporin resistant isolates were identified in this study. ESBL genes (blaSHV, blaTEM, blaOXA, and blaCTX-M and ampC genes (blaMOX, blaFOX, blaMIR(ACT-1, blaDHA, blaCIT and blaACC were subsequently detected among the 15 isolates. The results showed that these strains were positive only for blaTEM, blaOXA, blaCTX-M, intI1, and intI2. Furthermore, pulsed-field gel electrophoresis (PFGE analysis showed that the 40 isolates formed different profiles, and the PFGE patterns of Xinjiang isolates were distinct from Yunnan and Shanghai isolates by one obvious, large, missing band. In summary, similarities in resistance patterns were observed in strains with the same PFGE pattern. Overall, the results supported the need for more prudent selection and use of antibiotics in China. We suggest that antibiotic susceptibility testing should be performed at the start of an outbreak, and antibiotic use should be restricted to severe Shigella cases, based on resistance pattern variations observed in different regions. The data obtained in the current study might help to develop a strategy for the treatment of infections caused by S. flexneri 1b in China.

  9. Development of a Novel Protein Microarray Method for Serotyping Salmonella enterica Strains

    Cai, H.Y.; Lu, L; Muckle, C A; Prescott, J F; Chen, S.


    An antibody microarray assay was developed for Salmonella serotyping based on the Kauffmann-White scheme. A model (8 by 15) array was constructed using 35 antibodies for identification of 20 common Salmonella serovars and evaluated using 117 target and 73 nontarget Salmonella strains. The assay allowed complete serovar identification of 86 target strains and partial identification of 30 target strains and allowed exclusion of the 73 nontarget strains from the target serovars.

  10. Genome Sequence of Foot-and-Mouth Disease Virus Serotype O Isolated from Morocco in 2015

    Wadsworth, J.; Gray, A.; Abouchoaib, N.; King, D. P.; Knowles, N. J.


    The genome of a virus isolated from an outbreak of foot-and-mouth disease (FMD) in Morocco in 2015 is described here. This virus is classified as lineage Ind-2001d within serotype O, topotype ME-SA (Middle East-South Asia). This lineage is endemic on the Indian subcontinent but has caused outbreaks in the Middle East and North Africa since 2013. PMID:27103736

  11. Complete Genome Sequence of Foot-and-Mouth Disease Virus Serotype O Isolated from Bangladesh

    Sultana, Munawar; Siddique, Mohammad Anwar; Momtaz, Samina; Rahman, Arafat; Ullah, Huzzat; Nandi, Shuvro Prokash; Hossain, M. Anwar


    Foot-and-mouth disease (FMD) is a highly infectious enzootic disease caused by FMD virus. The complete genome sequence of a circulatory FMD virus (FMDV) serotype O isolated from Natore, Bangladesh, is reported here. Genomic analysis revealed antigenic heterogeneity within the VP1 region, a fragment deletion, and insertions at the 5′ untranslated region (UTR) and 3A region compared to the genome of the available vaccine strain.

  12. A Case of Neck Abscess Caused by Salmonella Serotype D in a Patient with Liver Cirrhosis

    Kwon, Mee-Hye; Kang, Mi-Il; Chun, Ji-Young; Lim, Hyun-Woo; Yeum, Yoon-Sik; Kang, Young-Woo; Kim, Young-Jin; Kim, Young Keun


    Non-typhoidal salmonellosis, which is increasing nowadays in Korea as well as in the developed countries, is manifested as enteritis in most cases, but it also encompasses bacteremia, intraabdominal infections, and bone, joint and soft tissue infections. These rare diseases are known to result from primary gastrointestinal infection and subsequent bacteremia with or without symptoms. We experienced a case of neck abscess caused by Salmonella serotype D, which is a rare but important different...

  13. Rapid and early detection of Salmonella serotypes with hyperspectral microscopy and multivariate data analysis.

    Eady, Matthew; Park, Bosoon; Choi, Sun


    This study was designed to evaluate hyperspectral microscope images for early and rapid detection of Salmonella serotypes Enteritidis, Heidelberg, Infantis, Kentucky, and Typhimurium at incubation times of 6, 8, 10, 12, and 24 h. Images were collected by an acousto-optical tunable filter hyperspectral microscope imaging system with a metal halide light source measuring 89 contiguous wavelengths every 4 nm between 450 and 800 nm. Pearson correlation values were calculated for incubation times of 8, 10, and 12 h and compared with data for 24 h to evaluate the change in spectral signatures from bacterial cells over time. Regions of interest were analyzed at 30% of the pixels in an average cell size. Spectral data were preprocessed by applying a global data transformation algorithm and then subjected to principal component analysis (PCA). The Mahalanobis distance was calculated from PCA score plots for analyzing serotype cluster separation. Partial least-squares regression was applied for calibration and validation of the model, and soft independent modeling of class analogy was utilized to classify serotype clusters in the training set. Pearson correlation values indicate very similar spectral patterns for reduced incubation times ranging from 0.9869 to 0.9990. PCA score plots indicated cluster separation at all incubation times, with incubation time Mahalanobis distances of 2.146 to 27.071. Partial least-squares regression had a maximum root mean squared error of calibration of 0.0025 and a root mean squared error of validation of 0.0030. Soft independent modeling of class analogy correctly classified values at 8 h (98.32%), 10 h (96.67%), 12 h (88.33%), and 24 h (98.67%) with the optimal number of principal components (four or five). The results of this study suggest that Salmonella serotypes can be classified by applying a PCA to hyperspectral microscope imaging data from samples after only 8 h of incubation. PMID:25836390

  14. Adherence and ingestion of Escherichia coli serotype 055 by trophozoites of Entamoeba histolytica.

    Bracha, R; Mirelman, D.


    Carbohydrate-binding activity present on the Entamoeba histolytica cell surfaces was found to mediate the adherence of two types of bacteria, Escherichia coli serotype 055 and Salmonella greenside 050. Adherence was inhibited by low-molecular-weight carbohydrates (10 mg/ml) such as galactose, lactose, and N-acetylgalactosamine, as well as by asialofetuin and the lipopolysaccharide extracted from E. coli 055. Mild periodate oxidation of the bacteria inhibited their adherence, whereas heat inac...

  15. Culture confirmation of Escherichia coli serotype O157:H7 by direct immunofluorescence.

    Tison, D. L.


    An evaluation of a fluorescein-labeled, polyclonal, affinity-purified goat antibody to Escherichia coli serotype O157:H7 (Kirkegaard & Perry Laboratories Inc., Gaithersburg, Md.) was conducted to determine the efficacy of this research reagent for the rapid direct immunofluorescence identification of E. coli O157:H7 isolated from fecal specimens cultured on sorbitol-MacConkey (SMAC) agar. The E. coli O157:H7 fluorescent-antibody conjugate proved to be 100% sensitive and specific for the rapid...

  16. A Monoclonal Antibody Based Capture ELISA for Botulinum Neurotoxin Serotype B: Toxin Detection in Food

    Larry H. Stanker


    Full Text Available Botulism is a serious foodborne neuroparalytic disease, caused by botulinum neurotoxin (BoNT, produced by the anaerobic bacterium Clostridium botulinum. Seven toxin serotypes (A–H have been described. The majority of human cases of botulism are caused by serotypes A and B followed by E and F. We report here a group of serotype B specific monoclonal antibodies (mAbs capable of binding toxin under physiological conditions. Thus, they serve as capture antibodies for a sandwich (capture ELISA. The antibodies were generated using recombinant peptide fragments corresponding to the receptor-binding domain of the toxin heavy chain as immunogen. Their binding properties suggest that they bind a complex epitope with dissociation constants (KD’s for individual antibodies ranging from 10 to 48 × 10−11 M. Assay performance for all possible combinations of capture-detector antibody pairs was evaluated and the antibody pair resulting in the lowest level of detection (L.O.D., ~20 pg/mL was determined. Toxin was detected in spiked dairy samples with good recoveries at concentrations as low as 0.5 pg/mL and in ground beef samples at levels as low as 2 ng/g. Thus, the sandwich ELISA described here uses mAb for both the capture and detector antibodies (binding different epitopes on the toxin molecule and readily detects toxin in those food samples tested.

  17. Characterisation of antimicrobial resistance-associated integrons and mismatch repair gene mutations in Salmonella serotypes.

    Yang, Baowei; Zheng, Jie; Brown, Eric W; Zhao, Shaohua; Meng, Jianghong


    In this study, we examined the presence of integrons and Salmonella genomic island 1 (SGI1) and assessed their contribution to antimicrobial resistance as well as determining the extent of the mutator phenotype in Salmonella isolates. A total of 81 Salmonella enterica serotype Typhimurium isolates were examined for the presence of integrons and SGI1 and for hypermutators using polymerase chain reaction (PCR) and the mutator assay, respectively. An additional 336 Salmonella isolates were also used to screen for hypermutators. Fourteen S. Typhimurium isolates carried class 1 integrons, of which six were shown to possess SGI1. Five putative mutators, S. Typhimurium ST20751, S. enterica serotype Heidelberg 22396 and S. enterica serotype Enteritidis 17929, 17929N and 17929R, were identified among the 417 Salmonella isolates. Complementation analysis with the wild-type mutH, mutL, mutS and uvrD genes indicated that none of the five mutators contained defective mismatch repair (MMR) system alleles. DNA sequence analysis revealed that single point mutations resulting in aspartic acid (codon 87) substitution in the gyrA gene conferred resistance to nalidixic acid and/or other fluoroquinolone drugs (ciprofloxacin and enrofloxacin) among four isolates. Our findings indicated that integrons and SGI1 play an important role in multidrug resistance in Salmonella. The incidence of hypermutators owing to defective MMR in Salmonella appears to be rare. PMID:19013057

  18. Evidence for a new avian paramyxovirus serotype 10 detected in rockhopper penguins from the Falkland Islands.

    Miller, Patti J; Afonso, Claudio L; Spackman, Erica; Scott, Melissa A; Pedersen, Janice C; Senne, Dennis A; Brown, Justin D; Fuller, Chad M; Uhart, Marcela M; Karesh, William B; Brown, Ian H; Alexander, Dennis J; Swayne, David E


    The biological, serological, and genomic characterization of a paramyxovirus recently isolated from rockhopper penguins (Eudyptes chrysocome) suggested that this virus represented a new avian paramyxovirus (APMV) group, APMV10. This penguin virus resembled other APMVs by electron microscopy; however, its viral hemagglutination (HA) activity was not inhibited by antisera against any of the nine defined APMV serotypes. In addition, antiserum generated against this penguin virus did not inhibit the HA of representative viruses of the other APMV serotypes. Sequence data produced using random priming methods revealed a genomic structure typical of APMV. Phylogenetic evaluation of coding regions revealed that amino acid sequences of all six proteins were most closely related to APMV2 and APMV8. The calculation of evolutionary distances among proteins and distances at the nucleotide level confirmed that APMV2, APMV8, and the penguin virus all were sufficiently divergent from each other to be considered different serotypes. We propose that this isolate, named APMV10/penguin/Falkland Islands/324/2007, be the prototype virus for APMV10. Because of the known problems associated with serology, such as antiserum cross-reactivity and one-way immunogenicity, in addition to the reliance on the immune response to a single protein, the hemagglutinin-neuraminidase, as the sole base for viral classification, we suggest the need for new classification guidelines that incorporate genome sequence comparisons. PMID:20702635

  19. Phylogenetic analysis of bluetongue virus serotype 4 field isolates from Argentina.

    Legisa, D; Gonzalez, F; De Stefano, G; Pereda, A; Dus Santos, M J


    Bluetongue is an insect-transmitted viral disease of ruminant species, which represents a major barrier to the international trade of animals and their products. Bluetongue virus (BTV) has a genome composed of ten linear segments of dsRNA, which code for at least ten different viral proteins. In South America, serological evidence for the presence of BTV has been found in Peru, Argentina, Brazil, Ecuador and Chile. Brazil and Argentina are the only South American countries where BTV has been isolated. In Brazil, only one BTV isolate, serotype 12, has been reported, whereas in Argentina five BTV serotype 4 isolates have been obtained from cattle without clinical signs. Three of these five isolates were isolated during 1999-2001, whereas two of them were obtained as part of the present work. This study describes sequence comparisons and phylogenetic analyses of segment (Seg)-2, Seg-3, Seg-6, Seg-7 and Seg-10 of the first Argentinian field isolates of BTV. The analysis of Seg-2 and Seg-6 resulted in a single cluster of Argentinian sequences into the serotype 4 clade. In addition, the Argentinian sequences grouped within the nucleotype A clade, along with reference strains. The analysis of Seg-3, Seg-7 and Seg-10 showed that the Argentinian isolates grouped into the western topotype, indicating that the circulating virus had an African/European origin. Phylogenetic analysis revealed that the Argentinian sequences present a South American genetic identity, suggesting an independent lineage evolution. PMID:23152367

  20. Serotype Distribution, Antimicrobial Susceptibility, and Molecular Epidemiology of Streptococcus pneumoniae Isolated from Children in Shanghai, China.

    Fen Pan

    Full Text Available Streptococcus pneumoniae is a common pathogenic cause of pediatric infections. This study investigated the serotype distribution, antimicrobial susceptibility, and molecular epidemiology of pneumococci before the introduction of conjugate vaccines in Shanghai, China.A total of 284 clinical pneumococcal isolates (270, 5, 4,3, and 2 of which were isolated from sputum, bronchoalveolar lavage fluid, blood, cerebral spinal fluid, and ear secretions, respectively from children less than 14 years of age who had not been vaccinated with a conjugate vaccine, were collected between January and December in 2013. All isolates were serotyped by multiplex polymerase chain reaction or quellung reactions and antimicrobial susceptibility testing was performed using the broth microdilution method. The molecular epidemiology of S.pneumoniae was analyzed by multilocus sequence typing (MLST.Among the 284 pneumococcal isolates, 19F (33.5%, 19A (14.1%, 23F (12.0%, and 6A (8.8% were the most common serotypes and the coverage rates of the 7-, 10-, and 13-valent pneumococcal conjugate vaccines (PCV7, PCV10, and PCV13 were 58.6%, 59.4% and 85.1%, respectively. Antimicrobial susceptibility showed that the prevalence rates of S.pneumoniae resistance to penicillin were 11.3% (32/284. Approximately 88.0% (250/284 of the isolates exhibited multi-drug resistance. MLST analysis revealed a high level of diversity, with 65 sequence types (STs among 267 isolates. Specifically, the four predominant STs were ST271 (24.3%, 65/267, ST320 (11.2%, 30/267, ST81 (9.7%, 26/267, and ST3173 (5.2%, 14/267, which were mainly associated with serotypes 19F, 19A, 23F, and 6A, respectively.The prevalent serotypes among clinical isolates from children were 19F, 19A, 23F, and 6A and these isolates showed high resistance rates to β-lactams and macrolides. The Taiwan19F-14 clone played a predominant role in the dissemination of pneumococcal isolates in Shanghai, China. Therefore, continued and