157 results on '"Rycroft, Andrew"'
Search Results
2. Antimicrobial susceptibility and genetic profile of Mycoplasma hyopneumoniae isolates from Brazil
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Gonzaga, Natália Fialho, de Souza, Luiz Fernando Lino, Santos, Marcus Rebouças, Assao, Viviane Sisdelli, Rycroft, Andrew, Deeney, Alannah Saskia, Fietto, Juliana Lopes Rangel, Bressan, Gustavo Costa, Moreira, Maria Aparecida Scatamburlo, and Silva-Júnior, Abelardo
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- 2020
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3. Mycoplasma hyopneumoniae evades phagocytic uptake by porcine alveolar macrophages in vitro
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Deeney, Alannah S., Maglennon, Gareth A., Chapat, Ludivine, Crussard, Steve, Jolivet, Edmond, and Rycroft, Andrew N.
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- 2019
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4. Exposure of Cats in Southern Africa to Coxiella burnetii, the Agent of Q Fever
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Matthewman, Linda, Kelly, Patrick, Hayter, Deidre, Downie, Susan, Wray, Kylie, Bryson, Nigel, Rycroft, Andrew, and Raoult, Didier
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- 1997
5. Domestic Cats as Indicators of the Presence of Spotted Fever and Typhus Group Rickettsiae
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Matthewman, Linda, Kelly, Patrick, Hayter, Diane, Downie, Susan, Wray, Kylie, Bryson, Nigel, Rycroft, Andrew, and Raoult, Didier
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- 1997
6. Rationally designed mariner vectors for functional genomic analysis of Actinobacillus pleuropneumoniae and other Pasteurellaceae species by transposon-directed insertion-site sequencing (TraDIS)
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Bossé, Janine T, Li, Yanwen, Leanse, Leon G, Zhou, Liqing, Chaudhuri, Roy R, Peters, Sarah E, Wang, Jinhong, Maglennon, Gareth A, Holden, Matthew TG, Maskell, Duncan J, Tucker, Alexander W, Wren, Brendan W, Rycroft, Andrew N, Langford, Paul R, and BRaDP1T consortium
- Abstract
Comprehensive identification of conditionally essential genes requires efficient tools for generating high-density transposon libraries that, ideally, can be analysed using next-generation sequencing methods such as Transposon Directed Insertion-site Sequencing (TraDIS). The Himar1 (mariner) transposon is ideal for generating near-saturating mutant libraries, especially in AT-rich chromosomes, as the requirement for integration is a TA dinucleotide, and this transposon has been used for mutagenesis of a wide variety of bacteria. However, plasmids for mariner delivery do not necessarily work well in all bacteria. In particular, there are limited tools for functional genomic analysis of Pasteurellaceae species of major veterinary importance, such as swine and cattle pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida, respectively. Here, we developed plasmids, pTsodCPC9 and pTlacPC9 (differing only in the promoter driving expression of the transposase gene), that allow delivery of mariner into both these pathogens, but which should also be applicable to a wider range of bacteria. Using the pTlacPC9 vector, we have generated, for the first time, saturating mariner mutant libraries in both A. pleuropneumoniae and P. multocida that showed a near random distribution of insertions around the respective chromosomes as detected by TraDIS. A preliminary screen of 5000 mutants each identified 8 and 14 genes, respectively, that are required for growth under anaerobic conditions. Future high-throughput screening of the generated libraries will facilitate identification of mutants required for growth under different conditions, including in vivo, highlighting key virulence factors and pathways that can be exploited for development of novel therapeutics and vaccines.
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- 2021
7. Necrosis from needlestick injury with live porcine vaccine
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Rycroft, Andrew N, Assavacheep, Pornchalit, Jacobs, Michael, and Langford, Paul R
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- 2011
8. Regulation of pga operon expression and biofilm formation in Actinobacillus pleuropneumoniae by [[sigma].sup.E] and H-NS
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Bosse, Janine T., Sinha, Sunita, Li, Ming-Shi, O'Dwyer, Cliona A., Nash, John H.E., Rycroft, Andrew N., Kroll, J. Simon, and Langford, Paul R.
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Actinobacillus -- Genetic aspects ,Actinobacillus -- Physiological aspects ,Operons -- Physiological aspects ,Gene expression -- Physiological aspects ,Genetic regulation -- Physiological aspects ,Microbial mats -- Growth ,Microbial mats -- Genetic aspects ,Company growth ,Biological sciences - Abstract
Clinical isolates of the porcine pathogen Actinobacillus pleuropneumoniae often form adherent colonies on agar plates due to expression of an operon, pgaABCD, encoding a poly-[beta]-1,6-N-acetyl-D-glucosamine (PGA) extracellular matrix. The adherent colony phenotype, which correlates with the ability to form biofilms on the surfaces of polystyrene plates, is lost following serial passage in broth culture, and repeated passage of the nonadherent variants on solid media does not result in reversion to the adherent colony phenotype. In order to investigate the regulation of PGA expression and biofilm formation in A. pleuropneumoniae, we screened a bank of transposon mutants of the nonadherent serovar 1 strain [S4074.sup.T] and identified mutations in two genes, rseA and hns, which resulted in the formation of the adherent colony phenotype. In other bacteria, including the Enterobacteriaceae, H-NS acts as a global gene regulator, and RseA is a negative regulator of the extracytoplasmic stress response sigma factor [[sigma].sup.E]. Transcription profiling of A. pleuropneumoniae rseA and hns mutants revealed that both [[sigma].sup.E] and H-NS independently regulate expression of the pga operon. Transcription of the pga operon is initiated from a [[sigma].sup.E] promoter site in the absence of H-NS, and upregulation of [[sigma].sup.E] is sufficient to displace H-NS, allowing transcription to proceed. In A. pleuropneumoniae, H-NS does not act as a global gene regulator but rather specifically regulates biofilm formation via repression of the pga operon. Positive regulation of the pga operon by [[sigma].sup.E] indicates that biofilm formation is part of the extracytoplasmic stress response in A. pleuropneumoniae. doi: 10.1128/JB.01513-09
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- 2010
9. Draft Genome Sequences of the Type Strains of Actinobacillus indolicus (46K2C) and Actinobacillus porcinus (NM319), Two NAD-Dependent Bacterial Species Found in the Respiratory Tract of Pigs
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Bossé, Janine T, Li, Yanwen, Fernandez Crespo, Roberto, Angen, Øystein, Holden, Matthew TG, Weinert, Lucy A, Maskell, Duncan J, Tucker, Alexander W, Wren, Brendan W, Rycroft, Andrew N, Langford, Paul R, BRaDP1T consortium, University of St Andrews. School of Medicine, University of St Andrews. Biomedical Sciences Research Complex, University of St Andrews. Infection and Global Health Division, University of St Andrews. Infection Group, Bossé, Janine T [0000-0002-8491-4779], Holden, Matthew TG [0000-0002-4958-2166], Langford, Paul R [0000-0002-6368-4724], Apollo - University of Cambridge Repository, Pfizer Limited (UK), and Biotechnology and Biological Sciences Research Council (BBSRC)
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PLEUROPNEUMONIAE ,education ,Nad dependent ,QH426 Genetics ,Biology ,Lung pathology ,Microbiology ,Genome ,03 medical and health sciences ,BRaDP1T consortium ,Immunology and Microbiology (miscellaneous) ,stomatognathic system ,Family pasteurellaceae ,Genetics ,Actinobacillus porcinus ,medicine ,Molecular Biology ,QH426 ,FAMILY PASTEURELLACEAE ,health care economics and organizations ,030304 developmental biology ,0303 health sciences ,Science & Technology ,030306 microbiology ,Actinobacillus indolicus ,Genome Sequences ,DAS ,QR Microbiology ,3. Good health ,QR ,stomatognathic diseases ,medicine.anatomical_structure ,Life Sciences & Biomedicine ,Respiratory tract - Abstract
We report here the draft genome sequences of the type strains of Actinobacillus indolicus (46K2C) and Actinobacillus porcinus (NM319). These NAD-dependent bacterial species are frequently found in the upper respiratory tract of pigs and are occasionally associated with lung pathology.
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- 2020
10. Publisher Correction: Genomic signatures of human and animal disease in the zoonotic pathogen Streptococcus suis
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Weinert, Lucy A, Chaudhuri, Roy R, Wang, Jinhong, Peters, Sarah E, Corander, Jukka, Jombart, Thibaut, Baig, Abiyad, Howell, Kate J, Vehkala, Minna, Välimäki, Niko, Harris, David, Chieu, Tran Thi Bich, Van Vinh Chau, Nguyen, Campbell, James, Schultsz, Constance, Parkhill, Julian, Bentley, Stephen D, Langford, Paul R, Rycroft, Andrew N, Wren, Brendan W, Farrar, Jeremy, Baker, Stephen, Hoa, Ngo Thi, Holden, Matthew TG, Tucker, Alexander W, Maskell, Duncan J, and BRaDP1T Consortium
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0303 health sciences ,Multidisciplinary ,biology ,030306 microbiology ,Science ,Animal disease ,General Physics and Astronomy ,Streptococcus suis ,General Chemistry ,biology.organism_classification ,General Biochemistry, Genetics and Molecular Biology ,3. Good health ,Microbiology ,03 medical and health sciences ,lcsh:Q ,lcsh:Science ,Zoonotic pathogen ,030304 developmental biology - Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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- 2019
11. Mycoplasmas associated with canine infectious respiratory disease
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Chalker, Victoria J., Owen, Wanda M.A., Paterson, Caren, Barker, Emily, Brooks, Harriet, Rycroft, Andrew N., and Brownlie, Joe
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Dogs -- Diseases ,Microbiology -- Research ,Mycoplasma infections -- Research ,Respiratory tract diseases -- Research ,Biological sciences - Abstract
Canine infectious respiratory disease (CIRD) is a complex infection that occurs worldwide predominantly in kennelled dogs, and several bacterial and viral micro-organisms have been associated with outbreaks of CIRD. However, few studies have comprehensively examined the species of mycoplasma present in healthy dogs and those with CIRD. As part of an extensive study investigating the micro-organisms involved in CIRD, the species of mycoplasma present throughout the respiratory tract of dogs with and without CIRD were determined. Mycoplasmas were cultured from tonsillar, tracheal and bronchial lavage samples, and identified to the species level by PCR and sequencing. Mycoplasma cynos was demonstrated on the ciliated tracheal epithelium by in situ hybridization and was the only mollicute found to be associated with CIRD, but only in the lower respiratory tract. Isolation of M. cynos was correlated with an increased severity of CIRD, younger age and a longer time in the kennel.
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- 2004
12. Necrosis from needlestick injury with live Actinobacillus pleuropneumoniae porcine vaccine
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Rycroft, Andrew N, Assavacheep, Pornchalit, Jacobs, Michael, and Langford, Paul R
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- 2011
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13. Generation and Evaluation of a Glaesserella (Haemophilus) parasuis Capsular Mutant
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Eberle, Kirsten C., primary, Hau, Samantha J., additional, Luan, Shi-Lu, additional, Weinert, Lucy A., additional, Stasko, Judith A., additional, Wang, Jinhong, additional, Peters, Sarah E., additional, Langford, Paul R., additional, Rycroft, Andrew N., additional, Wren, Brendan W., additional, Maskell, Duncan J., additional, Tucker, Alexander W., additional, and Brockmeier, Susan L., additional
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- 2020
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14. Antimicrobial susceptibility and genetic profile of Mycoplasma hyopneumoniae isolates from Brazil
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Gonzaga, Natália Fialho, primary, de Souza, Luiz Fernando Lino, additional, Santos, Marcus Rebouças, additional, Assao, Viviane Sisdelli, additional, Rycroft, Andrew, additional, Deeney, Alannah Saskia, additional, Fietto, Juliana Lopes Rangel, additional, Bressan, Gustavo Costa, additional, Moreira, Maria Aparecida Scatamburlo, additional, and Silva-Júnior, Abelardo, additional
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- 2019
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15. Phagocytosis by pig alveolar macrophages of actinobacillus pleuropneumoniae serotype 2 mutant strains defective in haemolysin II (ApxII) and pelurotoxin (ApxIII)
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Cullen, Janice M. and Rycroft, Andrew N.
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Macrophages -- Research ,Phagocytosis -- Research ,Biological sciences - Abstract
A study of pig alveolar macrophages reveals that the macrophages phagocytose Actinobacillus pleuropneumoniae HK 361 only in the presence of convalescent pig serum and absence of haemolysin II (ApxII) and pleurotoxin (ApxIII). Combination with ApxII exhibits extreme toxicity that the immune serum is unable to neutralize. The mutant form incubator with convalescent pig serum yields undamaged macrophages.
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- 1994
16. Comparative sequence analysis of the capsular polysaccharide loci of Actinobacillus pleuropneumoniae serovars 1-18, and development of two multiplex PCRs for comprehensive capsule typing
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Bossé, Janine T, Li, Yanwen, Fernandez Crespo, Roberto, Lacouture, Sonia, Gottschalk, Marcelo, Sárközi, Rita, Fodor, László, Casas Amoribieta, Maria, Angen, Øystein, Nedbalcova, Katerina, Holden, Matthew TG, Maskell, Duncan J, Tucker, Alexander W, Wren, Brendan W, Rycroft, Andrew N, Langford, Paul R, BRaDP1T consortium, Biotechnology and Biological Sciences Research Council (BBSRC), Pfizer Limited (UK), Biotechnology and Biological Sciences Research Council, University of St Andrews. School of Medicine, University of St Andrews. Infection and Global Health Division, University of St Andrews. Biomedical Sciences Research Complex, and University of St Andrews. Infection Group
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0301 basic medicine ,Serotype ,Swine ,QH301 Biology ,animal diseases ,Actinobacillus Infections ,Multiplex ,Pathogen ,Swine Diseases ,Genetics ,biology ,Actinobacillus pleuropneumoniae ,Polysaccharides, Bacterial ,QR Microbiology ,General Medicine ,Amplicon ,respiratory system ,3. Good health ,Pleuropneumonia ,Sequence Analysis ,mPCR ,0605 Microbiology ,Sequence analysis ,030106 microbiology ,Serovars ,Serogroup ,Microbiology ,Article ,QH301 ,03 medical and health sciences ,BRaDP1T consortium ,medicine ,Animals ,Typing ,Diagnostic ,Veterinary Sciences ,Serotyping ,Bacterial Capsules ,General Veterinary ,0707 Veterinary Sciences ,A. pleuropneumoniae ,Capsule typing ,DAS ,biology.organism_classification ,medicine.disease ,bacterial infections and mycoses ,respiratory tract diseases ,QR ,030104 developmental biology ,Multiplex Polymerase Chain Reaction - Abstract
Highlights • Analysis of complete capsule loci in all 18 serovars of A. pleuropneumoniae. • Novel insights into evolution of capsule loci in A. pleuropneumoniae. • Development of two mPCRs for comprehensive capsule typing., Problems with serological cross-reactivity have led to development of a number of PCRs (individual and multiplex) for molecular typing of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. Most of these assays were developed for detection of specific amplicons within capsule biosynthetic genes before the availability of complete sequences for the different serovars. Here we describe comparative analysis of the complete capsular loci for all 18 serovars of A. pleuropneumoniae, and development of two multiplex PCRs for comprehensive capsule typing of this important pig pathogen.
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- 2018
17. Proposal of serovars 17 and 18 of Actinobacillus pleuropneumoniae based on serological and genotypic analysis
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Bossé, Janine T, Li, Yanwen, Sárközi, Rita, Fodor, László, Lacouture, Sonia, Gottschalk, Marcelo, Casas Amoribieta, Maria, Angen, Øystein, Nedbalcova, Katerina, Holden, Matthew TG, Maskell, Duncan J, Tucker, Alexander W, Wren, Brendan W, Rycroft, Andrew N, Langford, Paul R, BRaDP1T consortium, Biotechnology and Biological Sciences Research Council (BBSRC), Pfizer Limited (UK), Biotechnology and Biological Sciences Research Council, University of St Andrews. School of Medicine, University of St Andrews. Infection and Global Health Division, University of St Andrews. Infection Group, and University of St Andrews. Biomedical Sciences Research Complex
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0301 basic medicine ,Serotype ,Swine ,Biovar ,animal diseases ,Denmark ,DIVERSITY ,Polymerase Chain Reaction ,Actinobacillus Infections ,Genotype ,ASSAY ,Diagnostics ,Genetics ,Swine Diseases ,biology ,Structural gene ,Actinobacillus pleuropneumoniae ,General Medicine ,3. Good health ,PCR ,Serovar 18 ,SEROTYPE-2 ,Serovar 17 ,QR355 Virology ,Life Sciences & Biomedicine ,MULTIPLEX PCR ,0605 Microbiology ,DNA, Bacterial ,Canada ,GENES ,030106 microbiology ,Locus (genetics) ,QH426 Genetics ,Serogroup ,Microbiology ,Article ,03 medical and health sciences ,BRaDP1T consortium ,Animals ,BIOSYNTHESIS ,Veterinary Sciences ,Serotyping ,CAPSULAR POLYSACCHARIDES ,Gene ,QH426 ,Bacterial Capsules ,DNA Primers ,Whole genome sequencing ,QR355 ,Science & Technology ,General Veterinary ,IDENTIFICATION ,Whole Genome Sequencing ,0707 Veterinary Sciences ,STRAINS ,DAS ,biology.organism_classification ,bacterial infections and mycoses ,Capsule genes - Abstract
Highlights • Identification of two new serovars of Actinobacillus pleuropneumoniae. • Serological confirmation of specific reactivity with homologous antisera. • Characterization of the capsule loci of serovars 17 and 18. • Development of PCRs for molecular diagnostics., The aim of this study was to investigate isolates of Actinobacillus pleuropneumoniae previously designated serologically either as non-typable (NT) or as ‘K2:07’, which did not produce serovar-specific amplicons in PCR assays. We used whole genome sequencing to identify the capsule (CPS) loci of six previously designated biovar 1 NT and two biovar 1 ‘K2:O7’ isolates of A. pleuropneumoniae from Denmark, as well as a recent biovar 2 NT isolate from Canada. All of the NT isolates have the same six-gene type I CPS locus, sharing common cpsABC genes with serovars 2, 3, 6, 7, 8, 9, 11 and 13. The two ‘K2:O7’ isolates contain a unique three-gene type II CPS locus, having a cpsA gene similar to that of serovars 1, 4, 12, 14 and 15. The previously NT isolates share the same O-antigen genes, found between erpA and rpsU, as serovars 3, 6, 8, and 15. Whereas the ‘K2:O7’ isolates, have the same O-antigen genes as serovar 7, which likely contributed to their previous mis-identification. All of the NT and ‘K2:O7’ isolates have only the genes required for production of ApxII (apxIICA structural genes, and apxIBD export genes). Rabbit polyclonal antisera raised against representative isolates with these new CPS loci demonstrated distinct reactivity compared to the 16 known serovars. The serological and genomic results indicate that the isolates constitute new serovars 17 (previously NT) and 18 (previously ‘K2:O7’). Primers designed for amplification of specific serovar 17 and 18 sequences for molecular diagnostics will facilitate epidemiological tracking of these two new serovars of A. pleuropneumoniae.
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- 2018
18. Pathotyping the Zoonotic Pathogen Streptococcus suis: Novel Genetic Markers To Differentiate Invasive Disease-Associated Isolates from Non-Disease-Associated Isolates from England and Wales
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Wileman, Thomas M., primary, Weinert, Lucy A., additional, Howell, Kate J., additional, Wang, Jinhong, additional, Peters, Sarah E., additional, Williamson, Susanna M., additional, Wells, Jerry M., additional, Langford, Paul R., additional, Rycroft, Andrew N., additional, Wren, Brendan W., additional, Maskell, Duncan J., additional, and Tucker, Alexander W., additional
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- 2019
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19. Patterns of antimicrobial resistance in Streptococcus suis isolates from pigs with or without streptococcal disease in England between 2009 and 2014
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Hernandez-Garcia, Juan, Wang, Jinhong, Restif, Olivier, Holmes, Mark A., Mather, Alison E., Weinert, Lucy A., Wileman, Thomas M., Thomson, Jill R., Langford, Paul R., Wren, Brendan W., Rycroft, Andrew, Maskell, Duncan J., and Tucker, Alexander W.
- Abstract
Antimicrobial resistance in Streptococcus suis, a global zoonotic pathogen of pigs, has been mostly studied only in diseased animals using surveys that have not evaluated changes over time. We compared patterns of resistance between S. suis isolates from clinical cases of disease (CC) and non-clinical case (NCC) pigs in England, collected over two discrete periods, 2009–2011 and 2013–2014. Minimum inhibitory concentrations (MIC) of 17 antimicrobials (nine classes) were determined on 405 S. suis isolates categorised by sampling period and disease association to assess changes in resistance over time and association with disease. First, isolates were characterized as resistant or susceptible using published clinical breakpoints. Second, epidemiological cut-offs (ECOFF) were derived from MIC values, and isolates classified as wild type (WT) below the ECOFF and non-wild type (NWT) above the ECOFF. Finally, isolate subsets were analysed for shifts in MIC distribution. NCC isolates were more resistant than CC isolates to cephalosporins, penams, pleuromutilins, potentiated sulphonamides and tetracyclines in both study periods. Resistance levels among CC isolates increased in 2013–2014 relative to 2009–2011 for antimicrobials including aminoglycosides, cephalosporins, fluoroquinolones, pleuromutilins, potentiated sulphonamides and tetracyclines. The prevalence of isolates categorised as NWT for five or more classes of antimicrobials was greater among NCC than CC isolates for both time periods, and increased with time. This study used standardised methods to identify significant shifts in antimicrobial resistance phenotypes of S. suis isolated from pigs in England, not only over time but also between isolates from known clinical cases or disease-free pigs.
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- 2017
20. Whole Genome Sequencing for Surveillance of Antimicrobial Resistance in Actinobacillus pleuropneumoniae
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Bossé, Janine T., Li, Yanwen, Rogers, Jon, Crespo, Roberto Fernandez, Li, Yinghui, Chaudhuri, Roy R., Holden, Matthew T. G., Maskell, Duncan J., Tucker, Alexander W., Wren, Brendan W., Rycroft, Andrew N., Langford, Paul R., BRaDP1T Consortium, University of St Andrews. School of Medicine, University of St Andrews. Infection Group, University of St Andrews. Infection and Global Health Division, University of St Andrews. Biomedical Sciences Research Complex, Maskell, Duncan [0000-0002-5065-653X], Tucker, Alexander [0000-0003-0062-0843], Apollo - University of Cambridge Repository, Biotechnology and Biological Sciences Research Council (BBSRC), and Pfizer Limited (UK)
- Subjects
0301 basic medicine ,PREDICTION ,QH301 Biology ,MANNHEIMIA-HAEMOLYTICA ,Respiratory tract ,SUSCEPTIBILITY ,Agar dilution ,BACTERIAL PATHOGENS ,chemistry.chemical_compound ,Ampicillin ,CONJUGATIVE ELEMENT ICE ,animal infections ,PIGS ,Tilmicosin ,Original Research ,biology ,HAEMOPHILUS-PARASUIS ,Animal infections ,Genomics ,QR Microbiology ,Antimicrobial ,Integrative conjugative elements ,PASTEURELLA-MULTOCIDA ,integrative conjugative elements ,Life Sciences & Biomedicine ,medicine.drug ,Plasmids ,Microbiology (medical) ,plasmids ,Tetracycline ,030106 microbiology ,Tylosin ,Microbiology ,03 medical and health sciences ,QH301 ,Enrofloxacin ,medicine ,genomics ,Actinobacillus pleuropneumoniae ,Science & Technology ,IDENTIFICATION ,DAS ,biology.organism_classification ,respiratory tract ,QR ,Antimicrobial resistance genes ,030104 developmental biology ,chemistry ,Pasteurellaceae ,antimicrobial resistance genes - Abstract
This work was supported by a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (BBSRC grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1, and BB/G018553/1), the UK Department for Environment, Food and Rural Affairs, and Zoetis (formerly Pfizer Animal Health) awarded to the Bacterial Respiratory Diseases of Pigs-1 Technology (BRaDP1T) consortium. MH was supported by the Wellcome Trust (grant number 098051). JR was funded from the former AHVLA’s Research and Development Internal Investment Fund (grant number RD0030c). The aim of this study was to evaluate the correlation between antimicrobial resistance (AMR) profiles of 96 clinical isolates of Actinobacillus pleuropneumoniae, an important porcine respiratory pathogen, and the identification of AMR genes in whole genome sequence (wgs) data. Susceptibility of the isolates to nine antimicrobial agents (ampicillin, enrofloxacin, erythromycin, florfenicol, sulfisoxazole, tetracycline, tilmicosin, trimethoprim, and tylosin) was determined by agar dilution susceptibility test. Except for the macrolides tested, elevated MICs were highly correlated to the presence of AMR genes identified in wgs data using ResFinder or BLASTn. Of the isolates tested, 57% were resistant to tetracycline [MIC ≥ 4 mg/L; 94.8% with either tet(B) or tet(H)]; 48% to sulfisoxazole (MIC ≥ 256 mg/L or DD = 6; 100% with sul2), 20% to ampicillin (MIC ≥ 4 mg/L; 100% with blaROB-1), 17% to trimethoprim (MIC ≥ 32 mg/L; 100% with dfrA14), and 6% to enrofloxacin (MIC ≥ 0.25 mg/L; 100% with GyrAS83F). Only 33% of the isolates did not have detectable AMR genes, and were sensitive by MICs for the antimicrobial agents tested. Although 23 isolates had MIC ≥ 32 mg/L for tylosin, all isolates had MIC ≥ 32 mg/L for tylosin, all isolates had MIC ≤ 16 mg/L for both erythromycin and tilmicosin, and no macrolide resistance genes or known point mutations were detected. Other than the GyrAS83F mutation, the AMR genes detected were mapped to potential plasmids. In addition to presence on plasmid(s), the tet(B) gene was also found chromosomally either as part of a 56 kb integrative conjugative element (ICEApl1) in 21, or as part of a Tn7 insertion in 15 isolates. Our results indicate that, with the exception of macrolides, wgs data can be used to accurately predict resistance of A. pleuropneumoniae to the tested antimicrobial agents and provides added value for routine surveillance. Publisher PDF
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- 2017
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21. A Unique Capsule Locus in the Newly Designated Actinobacillus pleuropneumoniae Serovar 16 and Development of a Diagnostic PCR Assay
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Bossé, Janine T., Li, Yanwen, Sárközi, Rita, Gottschalk, Marcelo, Angen, Øystein, Nedbalcova, Katerina, Rycroft, Andrew N., Fodor, László, Langford, Paul R., and Biotechnology and Biological Sciences Research Council (BBSRC)
- Subjects
DNA, Bacterial ,STRUCTURAL-CHARACTERIZATION ,Swine ,Serogroup ,Polymerase Chain Reaction ,Microbiology ,SEQUENCE ,Clinical Veterinary Microbiology ,Actinobacillus Infections ,diagnostics ,Animals ,LIPOPOLYSACCHARIDE-O-CHAIN ,Bacterial Capsules ,DNA Primers ,Swine Diseases ,Pleuropneumonia ,Science & Technology ,STRAINS ,Actinobacillus pleuropneumoniae ,Sequence Analysis, DNA ,11 Medical And Health Sciences ,06 Biological Sciences ,respiratory system ,serovar 16 ,PCR ,POLYSACCHARIDE ,Molecular Diagnostic Techniques ,Genetic Loci ,07 Agricultural And Veterinary Sciences ,Life Sciences & Biomedicine ,MULTIPLEX PCR ,Genome, Bacterial - Abstract
Actinobacillus pleuropneumoniae causes pleuropneumonia, an economically significant lung disease of pigs. Recently, isolates of A. pleuropneumoniae that were serologically distinct from the previously characterized 15 serovars were described, and a proposal was put forward that they comprised a new serovar, serovar 16. Here we used whole-genome sequencing of the proposed serovar 16 reference strain A-85/14 to confirm the presence of a unique capsular polysaccharide biosynthetic locus. For molecular diagnostics, primers were designed from the capsule locus of strain A-85/14, and a PCR was formulated that differentiated serovar 16 isolates from all 15 known serovars and other common respiratory pathogenic/commensal bacteria of pigs. Analysis of the capsule locus of strain A-85/14 combined with the previous serological data show the existence of a sixteenth serovar—designated serovar 16—of A. pleuropneumoniae.
- Published
- 2017
22. Glycosylation system investigation, Terra et al. Table S1 ;Glycosylation system investigation, Terra et al. Table S2;Glycosylation system investigation, Terra et al. Figure S1;Glycosylation system investigation, Terra et al. Figure S2 from The N-linking glycosylation system from Actinobacillus pleuropneumoniae is required for adhesion and has potential use in glycoengineering
- Author
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Cuccui, Jon, Terra, Vanessa S., Bossé, Janine T., Naegeli, Andreas, Abouelhadid, Sherif, Yanwen Li, Chia-Wei Lin, Prerna Vohra, Tucker, Alexander W., Rycroft, Andrew N., Maskell, Duncan J., Aebi, Markus, Langford, Paul R., and Wren, Brendan W.
- Subjects
2. Zero hunger - Abstract
Strains and plasmids used in this study; Primers used in this study for reverse transcriptase analysis of RNA extracts from A. pleuropneumoniae HS143, complementation of ngt and agt mutants and construction of A. pleuropneumoniae deletion mutants; Expression analyses of ngt and sodC. Reverse transcriptase PCR results of RNA extracted from A. pleuropneumoniae serotype 15 at various time points and in various mutants. 1, 1.5 hr cDNA template; 2, 1.5 hr RNA template; 3, 3.0 hr cDNA template; 4, 3.0 hr RNA; 5, 5.0 hr cDNA template; 6, 5.0 hr RNA template; 7, Δngt overnight culture cDNA template; 8, Δngt overnight culture RNA template; 9, Δngt complemented with pMKExpressNGT overnight culture cDNA template; 10, Δngt complemented with pMKExpressNGT overnight culture RNA template; 11, genomic DNA template (positive PCR control).; Predicted transcriptional promoter and Rho-independent terminators identified by bioinformatics analysis. V. Solovyev, A Salamov (2011) Automatic Annotation of Microbial Genomes and Metagenomic Sequences. In Metagenomics and its Applications in Agriculture, Biomedicine and Environmental Studies (Ed. R.W. Li), Nova Science Publishers, p. 61-78
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- 2017
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23. Evaluation of the recombinant proteins RlpB and VacJ as a vaccine for protection against Glaesserella parasuis in pigs.
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Hau, Samantha J., Luan, Shi-Lu, Loving, Crystal L., Nicholson, Tracy L., Wang, Jinhong, Peters, Sarah E., Seilly, David, Weinert, Lucy A., Langford, Paul R., Rycroft, Andrew N., Wren, Brendan W., Maskell, Duncan J., Tucker, Alexander W., and Brockmeier, Susan L.
- Subjects
ORGAN culture ,SWINE ,MEMBRANE proteins ,BACTERIAL antibodies ,BACTERIAL cell surfaces ,RECOMBINANT proteins - Abstract
Background: Glaesserella parasuis, the causative agent of Glӓsser's disease, is widespread in swine globally resulting in significant economic losses to the swine industry. Prevention of Glӓsser's disease in pigs has been plagued with an inability to design broadly protective vaccines, as many bacterin based platforms generate serovar or strain specific immunity. Subunit vaccines are of interest to provide protective immunity to multiple strains of G. parasuis. Selected proteins for subunit vaccination should be widespread, highly conserved, and surface exposed. Results: Two candidate proteins for subunit vaccination (RlpB and VacJ) against G. parasuis were identified using random mutagenesis and an in vitro organ culture system. Pigs were vaccinated with recombinant RlpB and VacJ, outer membrane proteins with important contributions to cellular function and viability. Though high antibody titers to the recombinant proteins and increased interferon-γ producing cells were found in subunit vaccinated animals, the pigs were not protected from developing systemic disease. Conclusions: It appears there may be insufficient RlpB and VacJ exposed on the bacterial surface for antibody to bind, preventing high RlpB and VacJ specific antibody titers from protecting animals from G. parasuis. Additionally, this work confirms the importance of utilizing the natural host species when assessing the efficacy of vaccine candidates. [ABSTRACT FROM AUTHOR]
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- 2020
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24. Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs
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Brockmeier, Susan L., primary, Loving, Crystal L., additional, Nicholson, Tracy L., additional, Wang, Jinhong, additional, Peters, Sarah E., additional, Weinert, Lucy, additional, Chaudhuri, Roy, additional, Seilly, David J., additional, Langford, Paul R., additional, Rycroft, Andrew, additional, Wren, Brendan W., additional, Maskell, Duncan J., additional, and Tucker, Alexander W., additional
- Published
- 2018
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25. In vivo testing of novel vaccine prototypes against Actinobacillus pleuropneumoniae
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Antenucci, Fabio, primary, Fougeroux, Cyrielle, additional, Deeney, Alannah, additional, Ørskov, Cathrine, additional, Rycroft, Andrew, additional, Holst, Peter Johannes, additional, and Bojesen, Anders Miki, additional
- Published
- 2018
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26. “Pathotyping” Multiplex PCR Assay for Haemophilus parasuis: a Tool for Prediction of Virulence
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Howell, Kate J., primary, Weinert, Lucy A., additional, Peters, Sarah E., additional, Wang, Jinhong, additional, Hernandez-Garcia, Juan, additional, Chaudhuri, Roy R., additional, Luan, Shi-Lu, additional, Angen, Øystein, additional, Aragon, Virginia, additional, Williamson, Susanna M., additional, Langford, Paul R., additional, Rycroft, Andrew N., additional, Wren, Brendan W., additional, Maskell, Duncan J., additional, and Tucker, Alexander W., additional
- Published
- 2017
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27. ICEApl1, an Integrative Conjugative Element Related to ICEHin1056, Identified in the Pig Pathogen Actinobacillus pleuropneumoniae
- Author
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Bosse, Janine T., Li, Yanwen, Crespo, Roberto Fernandez, Chaudhuri, Roy R., Rogers, Jon, Holden, Matthew T. G., Maskell, Duncan J., Tucker, Alexander W., Wren, Brendan W., Rycroft, Andrew N., Langford, Paul R., BRaDP1T Consortium, University of St Andrews. School of Medicine, University of St Andrews. Infection Group, and University of St Andrews. Biomedical Sciences Research Complex
- Subjects
Antibiotic resistance ,Conjugation ,NDAS ,Respiratory tract ,Animal infections ,QR Microbiology ,Pasteurellaceae ,R Medicine ,QR - Abstract
This work was supported by a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (BBSRC grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1, and BB/G018553/1), the UK Department for Environment, Food and Rural Affairs, and Zoetis (formerly Pfizer Animal Health) awarded to the Bacterial Respiratory Diseases of Pigs-1 Technology (BRaDP1T) Consortium. MTGH was supported by the Wellcome Trust (grant number 098051). JR was funded from the former AHVLA’s Research and Development Internal Investment Fund (grant number RD0030c). ICEApl1 was identified in the whole genome sequence of MIDG2331, a tetracycline resistant (MIC = 8 mg/L) serovar 8 clinical isolate of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. PCR amplification of virB4, one of the core genes involved in conjugation, was used to identify other A. pleuropneumoniae isolates potentially carrying ICEApl1. MICs for tetracycline were determined for virB4 positive isolates, and shotgun whole genome sequence analysis was used to confirm presence of the complete ICEApl1. The sequence of ICEApl1 is 56083 bp long and contains 67 genes including a Tn10 element encoding tetracycline resistance. Comparative sequence analysis was performed with similar integrative conjugative elements (ICEs) found in other members of the Pasteurellaceae. ICEApl1 is most similar to the 59393 bp ICEHin1056, from Haemophilus influenzae strain 1056. Although initially identified only in serovar 8 isolates of A. pleuropneumoniae (31 from the UK and 1 from Cyprus), conjugal transfer of ICEApl1 to representative isolates of other serovars was confirmed. All isolates carrying ICEApl1 had a MIC for tetracycline of 8 mg/L. This is, to our knowledge, the first description of an ICE in A. pleuropneumoniae, and the first report of a member of the ICEHin1056 subfamily in a non-human pathogen. ICEApl1 confers resistance to tetracycline, currently one of the more commonly used antibiotics for treatment and control of porcine pleuropneumonia. Publisher PDF
- Published
- 2016
28. Complete genome sequence of MIDG2331, a genetically tractable serovar 8 clinical isolate of Actinobacillus pleuropneumoniae
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Bossé, Janine T, Chaudhuri, Roy R., Li, Yanwen, Leanse, Leon G., Fernandez Crespo, Roberto, Coupland, Paul, Holden, Matthew T G, Bazzolli, Denise M., Maskell, Duncan J., Tucker, Alexander W., Wren, Brendan W., Rycroft, Andrew N., Langford, Paul R., BRaDP1T Consortium, University of St Andrews. School of Medicine, University of St Andrews. Infection Group, and University of St Andrews. Biomedical Sciences Research Complex
- Subjects
NDAS ,Prokaryotes ,QH426 Genetics ,R Medicine ,QH426 - Abstract
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/G019274/1, and BB/G018553/1. Biotech- nology and Biological Sciences Research Council (BBSRC) provided funding to Paul R Langford under grant number BB/K021109/1. MCTI | Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) provided funding to Denise Mara Soares Bazzolli under grant number PDE 201840/2011-1. This work was supported by a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1, and BB/G018553/1), the UK Department for Environment, Food and Rural Affairs, and Zoetis (formerly Pfizer Animal Health) awarded to the Bacterial Respiratory Dis- eases of Pigs-1 Technology (BRaDP1T) consortium, a grant from Con- selho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq; grant number PDE 201840/2011-1) awarded to D.M.B., and a BBSRC Imperial-Brazil partnering award (BB/K021109/1) awarded to P.R.L. M.T.G.H. and P.C. were supported by the Wellcome Trust (grant number 098051) We report here the complete annotated genome sequence of a clinical serovar 8 isolate Actinobacillus pleuropneumoniae MIDG2331. Unlike the serovar 8 reference strain 405, MIDG2331 is amenable to genetic manipulation via natural transformation as well as conjugation, making it ideal for studies of gene function. Publisher PDF
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- 2016
29. Complete genome sequence of MIDG2331, a genetically tractable serovar 8 clinical isolate of Actinobacillus pleuropneumoniae
- Author
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Crespo, Roberto Fernandez, Coupland, Paul, Bossé, Janine T., Chaudhuri, Roy R., Bazzolli, Denise M., Leanse, Leon G., Holden, Matthew T. G., Maskell, Duncan J., Tucker, Alexander W., Wren, Brendan W., Rycroft, Andrew N., and Langford, Paul R.
- Subjects
Actinobacillus pleuropneumoniae ,MIDG2331 - Abstract
We report here the complete annotated genome sequence of a clinical serovar 8 isolate Actinobacillus pleuropneumoniae MIDG2331. Unlike the serovar 8 reference strain 405, MIDG2331 is amenable to genetic manipulation via natural transformation as well as conjugation, making it ideal for studies of gene function.
- Published
- 2016
30. New plasmid tools for genetic analysis of Actinobacillus pleuropneumoniae and other Pasteurellaceae
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Bosse, Janine T., Durham, Andrew L., Rycroft, Andrew N., Kroll, J. Simon, and Langford, Paul R.
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Actinobacillus -- Genetic aspects ,Alcohol -- Environmental aspects ,Alcohol, Denatured -- Environmental aspects ,Gene expression -- Analysis ,Gene mutations -- Analysis ,Plasmids -- Research ,Genetic transcription -- Analysis ,Biological sciences - Abstract
The application of newly generated set of plasmids, based on the mobilizable shuttle vector pMIDG100 as an efficient tool for genetic manipulation of Actinobacillus pleuropneumoniae and other members of the Pasteurellaceae is reported. The ability of newly developed tandem reporter plasmid, pMC-tandem, carrying promoterless xylE and gfpmut3 genes, to detect transcriptional regulators is validated in A. pleuropneumoniae via use of the cloned rpoE ([sigma].sup.E]) promoter (P).
- Published
- 2009
31. A Unique Capsule Locus in the Newly Designated Actinobacillus pleuropneumoniae Serovar 16 and Development of a Diagnostic PCR Assay
- Author
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Bossé, Janine T., primary, Li, Yanwen, additional, Sárközi, Rita, additional, Gottschalk, Marcelo, additional, Angen, Øystein, additional, Nedbalcova, Katerina, additional, Rycroft, Andrew N., additional, Fodor, László, additional, and Langford, Paul R., additional
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- 2017
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32. Whole genome investigation of a divergent clade of the pathogen Streptococcus suis
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Baig, Abiyad, Weinert, Lucy A., Peters, Sarah E., Howell, Kate J., Chaudhuri, Roy R., Wang, Jinhong, Holden, Matthew T. G., Parkhill, Julian, Langford, Paul R., Rycroft, Andrew N., Wren, Brendan W., Tucker, Alexander W., Maskell, Duncan J., Biotechnology and Biological Sciences Research Council (BBSRC), University of St Andrews. School of Medicine, University of St Andrews. Infection Group, University of St Andrews. Biomedical Sciences Research Complex, Weinert, Lucy [0000-0002-9279-6012], Parkhill, Julian [0000-0002-7069-5958], Tucker, Alexander [0000-0003-0062-0843], Maskell, Duncan [0000-0002-5065-653X], and Apollo - University of Cambridge Repository
- Subjects
divergent ,Streptococcus suis ,capsule ,NDAS ,DIVERSITY ,lcsh:QR1-502 ,QH426 Genetics ,phylogeny ,Microbiology ,lcsh:Microbiology ,SDG 3 - Good Health and Well-being ,INFECTION ,GENUS STREPTOCOCCUS ,SWINE ,QH426 ,genome ,Original Research ,Science & Technology ,IDENTIFICATION ,SEROTYPES ,recombination ,virulence ,SP-NOV ,ORISRATTI ,Life Sciences & Biomedicine ,MLST - Abstract
This work was supported by a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1, and BB/G003203/1), the UK Department for Environment, Food and Rural Affairs and Zoetis, awarded to the Bacterial Respiratory Diseases of Pigs-1 Technology (BRaDP1T) consortium. Streptococcus suis is a major porcine and zoonotic pathogen responsible for significant economic losses in the pig industry and an increasing number of human cases. Multiple isolates of S. suis show marked genomic diversity. Here, we report the analysis of whole genome sequences of nine pig isolates that caused disease typical of S. suis and had phenotypic characteristics of S. suis, but their genomes were divergent from those of many other S. suis isolates. Comparison of protein sequences predicted from divergent genomes with those from normal S. suis reduced the size of core genome from 793 to only 397 genes. Divergence was clear if phylogenetic analysis was performed on reduced core genes and MLST alleles. Phylogenies based on certain other genes (16S rRNA, sodA, recN, and cpn60) did not show divergence for all isolates, suggesting recombination between some divergent isolates with normal S. suis for these genes. Indeed, there is evidence of recent recombination between the divergent and normal S. suis genomes for 249 of 397 core genes. In addition, phylogenetic analysis based on the 16S rRNA gene and 132 genes that were conserved between the divergent isolates and representatives of the broader Streptococcus genus showed that divergent isolates were more closely related to S. suis. Six out of nine divergent isolates possessed a S. suis-like capsule region with variation in capsular gene sequences but the remaining three did not have a discrete capsule locus. The majority (40/70), of virulence-associated genes in normal S. suis were present in the divergent genomes. Overall, the divergent isolates extend the current diversity of S. suis species but the phenotypic similarities and the large amount of gene exchange with normal S. suis gives insufficient evidence to assign these isolates to a new species or subspecies. Further, sampling and whole genome analysis of more isolates is warranted to understand the diversity of the species. Publisher PDF
- Published
- 2015
33. Characterisation of a mobilisable plasmid conferring florfenicol and chloramphenicol resistance in Actinobacillus pleuropneumoniae
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Bossé, Janine T, Li, Yanwen, Atherton, Tom G, Walker, Stephanie, Williamson, Susanna M, Rogers, Jon, Chaudhuri, Roy R, Weinert, Lucy A, Holden, Matthew TG, Maskell, Duncan J, Tucker, Alexander W, Wren, Brendan W, Rycroft, Andrew N, Langford, Paul R, BRaDP1T consortium, University of St Andrews. School of Medicine, University of St Andrews. Infection Group, and University of St Andrews. Biomedical Sciences Research Complex
- Subjects
Florfenicol ,Swine ,animal diseases ,Short Communication ,Molecular Sequence Data ,NDAS ,Chloramphenicol Resistance ,Microbiology ,chemistry.chemical_compound ,Actinobacillus Infections ,Plasmid ,Drug Resistance, Bacterial ,Animals ,Actinobacillus pleuropneumoniae ,Swine Diseases ,Thiamphenicol ,Base Sequence ,General Veterinary ,biology ,Pasteurellaceae ,Nucleic acid sequence ,Sequence Analysis, DNA ,General Medicine ,biology.organism_classification ,veterinary(all) ,stomatognathic diseases ,chemistry ,Pasteurellaceae Infections ,Plasmids - Abstract
Highlights • First complete sequence of a floR plasmid from Actinobacillus pleuropneumoniae • Extended similarity to floR plasmids in other Pasteurellaceae species • Conjugal transfer between between species confirmed, The complete nucleotide sequence of a 7.7 kb mobilisable plasmid (pM3446F), isolated from a florfenicol resistant isolate of Actinobacillus pleuropneumoniae, showed extended similarity to plasmids found in other members of the Pasteurellaceae containing the floR gene as well as replication and mobilisation genes. Mobilisation into other Pasteurellaceae species confirmed that this plasmid can be transferred horizontally.
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- 2015
34. Erratum: Genomic signatures of human and animal disease in the zoonotic pathogen Streptococcus suis
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Weinert, Lucy A., Chaudhuri, Roy R., Wang, Jinhong, Peters, Sarah E., Corander, Jukka, Jombart, Thibaut, Baig, Abiyad, Howell, Kate J., Vehkala, Minna, Välimäki, Niko, Harris, David, Chieu, Tran Thi Bich, Van Vinh Chau, Nguyen, Campbell, James, Schultsz, Constance, Parkhill, Julian, Bentley, Stephen D., Langford, Paul R., Rycroft, Andrew N., Wren, Brendan W., Farrar, Jeremy, Baker, Stephen, Thi Hoa, Ngo, Holden, Matthew T.G., Tucker, Alexander W., Maskell, Duncan J., and Medical Research Council (MRC)
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Multidisciplinary Sciences ,BRaDP1T Consortium ,Science & Technology ,Multidisciplinary ,MD Multidisciplinary ,Science & Technology - Other Topics ,General Physics and Astronomy ,General Chemistry ,Erratum ,General Biochemistry, Genetics and Molecular Biology - Published
- 2015
35. Antimicrobial susceptibility and genetic profile of Mycoplasma hyopneumoniaeisolates from Brazil
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Gonzaga, Natália Fialho, de Souza, Luiz Fernando Lino, Santos, Marcus Rebouças, Assao, Viviane Sisdelli, Rycroft, Andrew, Deeney, Alannah Saskia, Fietto, Juliana Lopes Rangel, Bressan, Gustavo Costa, Moreira, Maria Aparecida Scatamburlo, and Silva-Júnior, Abelardo
- Abstract
Mycoplasma hyopneumoniaeis the etiologic agent of porcine enzootic pneumonia, responsible for major production losses worldwide. The bacteria have a limited metabolism and need to obtain molecules from the growth environment, which causes multiple difficulties for in vitro culture. These limitations have a negative influence on the ability to carry out research for the development of the rational use of antimicrobials and vaccines. The objective of this investigation was to evaluate the genetic profile and in vitro susceptibility of field isolates of M. hyopneumoniaeto different antimicrobials. All 16 isolates obtained from the samples presented 100% of identity in the partial sequence of 16S rRNA gene when compared to M. hyopneumoniae. A dendrogram was created using the PCR results of the genes related to pathogenicity, and the isolates were distributed into four clusters, suggesting genetic variability among four different isolates circulating on the same farm. The minimum inhibitory concentration of the isolates was higher for the antimicrobials tylosin (< 0.001–16 mg/L) and spiramycin (< 0.001–16 mg/L) than for enrofloxacin (< 0.001–0.125 mg/L) and tiamulin (< 0.001–0.125 mg/L). Our results demonstrate the genetic variability among M. hyopneumoniaeisolates from pigs of the same farm, with differences in their susceptibility to antimicrobial agents.
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- 2020
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36. The use of genome wide association methods to investigate pathogenicity, population structure and serovar in Haemophilus parasuis
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Howell, Kate J, Weinert, Lucy A, Chaudhuri, Roy R, Luan, Shi-Lu, Peters, Sarah E, Corander, Jukka, Harris, David, Angen, Øystein, Aragon, Virginia, Bensaid, Albert, Williamson, Susanna M, Parkhill, Julian, Langford, Paul R, Rycroft, Andrew N, Wren, Brendan W, Holden, Matthew TG, Tucker, Alexander W, Maskell, Duncan J, BRADP1T Consortium, Weinert, Lucy [0000-0002-9279-6012], Parkhill, Julian [0000-0002-7069-5958], Tucker, Alexander [0000-0003-0062-0843], Maskell, Duncan [0000-0002-5065-653X], and Apollo - University of Cambridge Repository
- Subjects
Recombination, Genetic ,Haemophilus parasuis ,Virulence ,Swine ,Animals ,Genome, Viral ,Genome-Wide Association Study - Abstract
BACKGROUND: Haemophilus parasuis is the etiologic agent of Glässer's disease in pigs and causes devastating losses to the farming industry. Whilst some hyper-virulent isolates have been described, the relationship between genetics and disease outcome has been only partially established. In particular, there is weak correlation between serovar and disease phenotype. We sequenced the genomes of 212 isolates of H. parasuis and have used this to describe the pan-genome and to correlate this with clinical and carrier status, as well as with serotype. RESULTS: Recombination and population structure analyses identified five groups with very high rates of recombination, separated into two clades of H. parasuis with no signs of recombination between them. We used genome-wide association methods including discriminant analysis of principal components (DAPC) and generalised linear modelling (glm) to look for genetic determinants of this population partition, serovar and pathogenicity. We were able to identify genes from the accessory genome that were significantly associated with phenotypes such as potential serovar specific genes including capsule genes, and 48 putative virulence factors that were significantly different between the clinical and non-clinical isolates. We also show that the presence of many previously suggested virulence factors is not an appropriate marker of virulence. CONCLUSIONS: These genes will inform the generation of new molecular diagnostics and vaccines, and refinement of existing typing schemes and show the importance of the accessory genome of a diverse species when investigating the relationship between genotypes and phenotypes.
- Published
- 2014
37. The generation of successive unmarked mutations and chromosomal insertion of heterologous genes in Actinobacillus pleuropneumoniae using natural transformation
- Author
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Bossé, Janine T., Soares-Bazzolli, Denise M., Li, Yanwen, Wren, Brendan W., Tucker, Alexander W., Maskell, Duncan J., Rycroft, Andrew N., and Langford, Paul R.
- Subjects
Natural transformation ,Actinobacillus pleuropneumoniae - Abstract
We have developed a simple method of generating scarless, unmarked mutations in Actinobacillus pleuropneumoniae by exploiting the ability of this bacterium to undergo natural transformation, and with no need to introduce plasmids encoding recombinases or resolvases. This method involves two successive rounds of natural transformation using linear DNA: the first introduces a cassette carrying cat (which allows selection by chloramphenicol) and sacB (which allows counter-selection using sucrose) flanked by sequences to either side of the target gene; the second transformation utilises the flanking sequences ligated directly to each other in order to remove the cat-sacB cassette. In order to ensure efficient uptake of the target DNA during transformation, A. pleuropneumoniae uptake sequences are added into the constructs used in both rounds of transformation. This method can be used to generate multiple successive deletions and can also be used to introduce targeted point mutations or insertions of heterologous genes into the A. pleuropneumoniae chromosome for development of live attenuated vaccine strains. So far, we have applied this method to highly transformable isolates of serovars 8 (MIDG2331), which is the most prevalent in the UK, and 15 (HS143). By screening clinical isolates of other serovars, it should be possible to identify other amenable strains.
- Published
- 2014
38. Multiplex PCR assay for unequivocal differentiation of Actinobacillus pleuropneumoniae serovars 1 to 3, 5 to 8, 10, and 12
- Author
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Bossé, Janine T, Li, Yanwen, Angen, Oystein, Weinert, Lucy A, Chaudhuri, Roy R, Holden, Matt T, Williamson, Susanna M, Maskell, Duncan J, Tucker, Alexander W, Wren, Brendan W, Rycroft, Andrew N, Langford, Paul R, BRaDP1T Consortium, Holden, Matthew, University of St Andrews. School of Medicine, University of St Andrews. Infection Group, and University of St Andrews. Biomedical Sciences Research Complex
- Subjects
DNA, Bacterial ,Actinobacillus Infections ,Actinobacillus pleuropneumoniae ,Molecular Sequence Data ,Animals ,Humans ,QR Microbiology ,Sequence Analysis, DNA ,Serogroup ,Multiplex Polymerase Chain Reaction ,QR ,DNA Primers ,Clinical Veterinary Microbiology - Abstract
This work was supported by a Longer and Larger (LoLa) grant from the Biotechnology and Biological Sciences Research Council (grant numbers BB/G020744/1, BB/G019177/1, BB/G019274/1, and BB/G003203/1) and the United Kingdom Department for Environment, Food and Rural Af-fairs and Zoetis awarded to the Bacterial Respiratory Diseases of Pigs-1 Technology(BRaDP1T)consortium. An improved multiplex PCR, using redesigned primers targeting the serovar 3 capsule locus, which differentiates serovar 3, 6 and 8 Actinobacillus pleuropneumoniae isolates is described. The new primers eliminate an aberrant serovar 3 indicative amplicon found in some serovar 6 clinical isolates. Furthermore, we have developed a new multiplex PCR for detection of serovars 1-3, 5-8, 10, and 12 along with apxIV, thus extending the utility of this diagnostic PCR to cover a broader range of isolates. Publisher PDF
- Published
- 2014
39. The generation of successive unmarked mutations and chromosomal insertion of heterologous genes in Actinobacillus pleuropneumoniae using natural transformation
- Author
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Bossé, Janine T, Soares-Bazzolli, Denise M, Li, Yanwen, Wren, Brendan W, Tucker, Alexander W, Maskell, Duncan J, Rycroft, Andrew N, Langford, Paul R, and BRaDP1T Consortium
- Subjects
lcsh:R ,lcsh:Medicine ,lcsh:Q ,lcsh:Science - Abstract
We have developed a simple method of generating scarless, unmarked mutations in Actinobacillus pleuropneumoniae by exploiting the ability of this bacterium to undergo natural transformation, and with no need to introduce plasmids encoding recombinases or resolvases. This method involves two successive rounds of natural transformation using linear DNA: the first introduces a cassette carrying cat (which allows selection by chloramphenicol) and sacB (which allows counter-selection using sucrose) flanked by sequences to either side of the target gene; the second transformation utilises the flanking sequences ligated directly to each other in order to remove the cat-sacB cassette. In order to ensure efficient uptake of the target DNA during transformation, A. pleuropneumoniae uptake sequences are added into the constructs used in both rounds of transformation. This method can be used to generate multiple successive deletions and can also be used to introduce targeted point mutations or insertions of heterologous genes into the A. pleuropneumoniae chromosome for development of live attenuated vaccine strains. So far, we have applied this method to highly transformable isolates of serovars 8 (MIDG2331), which is the most prevalent in the UK, and 15 (HS143). By screening clinical isolates of other serovars, it should be possible to identify other amenable strains.
- Published
- 2014
40. Complete Genome Sequence of MIDG2331, a Genetically Tractable Serovar 8 Clinical Isolate of Actinobacillus pleuropneumoniae
- Author
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Bossé, Janine T., primary, Chaudhuri, Roy R., additional, Li, Yanwen, additional, Leanse, Leon G., additional, Fernandez Crespo, Roberto, additional, Coupland, Paul, additional, Holden, Matthew T. G., additional, Bazzolli, Denise M., additional, Maskell, Duncan J., additional, Tucker, Alexander W., additional, Wren, Brendan W., additional, Rycroft, Andrew N., additional, and Langford, Paul R., additional
- Published
- 2016
- Full Text
- View/download PDF
41. Development of a Multiplex PCR Assay for Rapid Molecular Serotyping of Haemophilus parasuis
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Howell, Kate J., primary, Peters, Sarah E., additional, Wang, Jinhong, additional, Hernandez-Garcia, Juan, additional, Weinert, Lucy A., additional, Luan, Shi-Lu, additional, Chaudhuri, Roy R., additional, Angen, Øystein, additional, Aragon, Virginia, additional, Williamson, Susanna M., additional, Parkhill, Julian, additional, Langford, Paul R., additional, Rycroft, Andrew N., additional, Wren, Brendan W., additional, Maskell, Duncan J., additional, and Tucker, Alexander W., additional
- Published
- 2015
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42. Development of a self-replicating plasmid system for Mycoplasma hyopneumoniae
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Maglennon, Gareth A, Cook, Beth S, Matthews, Dominic, Deeney, Alannah S, Bossé, Janine T, Langford, Paul R, Maskell, Duncan J, Tucker, Alexander W, Wren, Brendan W, Rycroft, Andrew N, and BRaDP1T consortium
- Abstract
Mycoplasma hyopneumoniae is a prevalent swine respiratory pathogen that is a major cause of economic loss to pig producers. Control is achieved by a combination of antimicrobials, vaccination and management practices, but current vaccines offer only partial control and there is a need for improved preventative strategies. A major barrier to advances in understanding the pathogenesis of M. hyopneumoniae and in developing new vaccines is the lack of tools to genetically manipulate the organism. We describe the development and optimisation of the first successful plasmid-based system for the genetic manipulation of M. hyopneumoniae. Our artificial plasmids contain the origin of replication (oriC) of M. hyopneumoniae along with tetM, conferring resistance to tetracycline. With these plasmids, we have successfully transformed M. hyopneumoniae strain 232 by electroporation, generating tetracycline resistant organisms. The persistence of extrachromosomal plasmid and maintenance of plasmid DNA over serial passages shows that these artificial plasmids are capable of self-replication in M. hyopneumoniae. In addition to demonstrating the amenability of M. hyopneumoniae to genetic manipulation and in optimising the conditions necessary for successful transformation, we have used this system to determine the minimum functional oriC of M. hyopneumoniae. In doing so, we have developed a plasmid with a small oriC that is stably maintained over multiple passages that may be useful in generating targeted gene disruptions. In conclusion, we have generated a set of plasmids that will be valuable in studies of M. hyopneumoniae pathogenesis and provide a major step forward in the study of this important swine pathogen.
- Published
- 2013
43. Transposon mutagenesis in Mycoplasma hyopneumoniae using a novel mariner-based system for generating random mutations
- Author
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Maglennon, Gareth A, primary, Cook, Beth S, additional, Deeney, Alannah S, additional, Bossé, Janine T, additional, Peters, Sarah E, additional, Langford, Paul R, additional, Maskell, Duncan J, additional, Tucker, Alexander W, additional, Wren, Brendan W, additional, and Rycroft, Andrew N, additional
- Published
- 2013
- Full Text
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44. A Unique Capsule Locus in the Newly Designated Actinobacillus pleuropneumoniaeSerovar 16 and Development of a Diagnostic PCR Assay
- Author
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Bossé, Janine T., Li, Yanwen, Sárközi, Rita, Gottschalk, Marcelo, Angen, Øystein, Nedbalcova, Katerina, Rycroft, Andrew N., Fodor, László, and Langford, Paul R.
- Abstract
ABSTRACTActinobacillus pleuropneumoniaecauses pleuropneumonia, an economically significant lung disease of pigs. Recently, isolates of A. pleuropneumoniaethat were serologically distinct from the previously characterized 15 serovars were described, and a proposal was put forward that they comprised a new serovar, serovar 16. Here we used whole-genome sequencing of the proposed serovar 16 reference strain A-85/14 to confirm the presence of a unique capsular polysaccharide biosynthetic locus. For molecular diagnostics, primers were designed from the capsule locus of strain A-85/14, and a PCR was formulated that differentiated serovar 16 isolates from all 15 known serovars and other common respiratory pathogenic/commensal bacteria of pigs. Analysis of the capsule locus of strain A-85/14 combined with the previous serological data show the existence of a sixteenth serovar—designated serovar 16—of A. pleuropneumoniae.
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- 2017
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45. Regulation of pga Operon Expression and Biofilm Formation in Actinobacillus pleuropneumoniae by σ E and H-NS
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Bossé, Janine T., primary, Sinha, Sunita, additional, Li, Ming-Shi, additional, O'Dwyer, Clíona A., additional, Nash, John H. E., additional, Rycroft, Andrew N., additional, Kroll, J. Simon, additional, and Langford, Paul R., additional
- Published
- 2010
- Full Text
- View/download PDF
46. Specific Strains of Escherichia coli Are Pathogenic for the Endometrium of Cattle and Cause Pelvic Inflammatory Disease in Cattle and Mice
- Author
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Sheldon, I. Martin, primary, Rycroft, Andrew N., additional, Dogan, Belgin, additional, Craven, Melanie, additional, Bromfield, John J., additional, Chandler, Alyssa, additional, Roberts, Mark H., additional, Price, Sian B., additional, Gilbert, Robert O., additional, and Simpson, Kenneth W., additional
- Published
- 2010
- Full Text
- View/download PDF
47. Pasteurellaceae ComE1 Proteins Combine the Properties of Fibronectin Adhesins and DNA Binding Competence Proteins
- Author
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Mullen, Lisa M., primary, Bossé, Janine T., additional, Nair, Sean P., additional, Ward, John M., additional, Rycroft, Andrew N., additional, Robertson, Giles, additional, Langford, Paul R., additional, and Henderson, Brian, additional
- Published
- 2008
- Full Text
- View/download PDF
48. Novel Adhesin from Pasteurella multocida That Binds to the Integrin-Binding Fibronectin FnIII 9-10 Repeats
- Author
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Mullen, Lisa M., primary, Nair, Sean P., additional, Ward, John M., additional, Rycroft, Andrew N., additional, Williams, Rachel J., additional, Robertson, Giles, additional, Mordan, Nicky J., additional, and Henderson, Brian, additional
- Published
- 2008
- Full Text
- View/download PDF
49. The use of genome wide association methods to investigate pathogenicity, population structure and serovar in Haemophilus parasuis.
- Author
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Howell, Kate J., Weinert, Lucy A., Chaudhuri, Roy R., Shi-Lu Luan, Peters, Sarah E., Corander, Jukka, Harris, David, Angen, Øystein, Aragon, Virginia, Bensaid, Albert, Williamson, Susanna M., Parkhill, Julian, Langford, Paul R., Rycroft, Andrew N., Wren, Brendan W., Holden, Matthew T. G., Tucker, Alexander W., and Maskell, Duncan J.
- Subjects
PATHOGENICITY of enteroviruses ,SWINE industry ,GENETIC recombination ,DISCRIMINANT analysis ,MICROBIAL virulence - Abstract
Background: Haemophilus parasuis is the etiologic agent of Glässer's disease in pigs and causes devastating losses to the farming industry. Whilst some hyper-virulent isolates have been described, the relationship between genetics and disease outcome has been only partially established. In particular, there is weak correlation between serovar and disease phenotype. We sequenced the genomes of 212 isolates of H. parasuis and have used this to describe the pan-genome and to correlate this with clinical and carrier status, as well as with serotype. Results: Recombination and population structure analyses identified five groups with very high rates of recombination, separated into two clades of H. parasuis with no signs of recombination between them. We used genome-wide association methods including discriminant analysis of principal components (DAPC) and generalised linear modelling (glm) to look for genetic determinants of this population partition, serovar and pathogenicity. We were able to identify genes from the accessory genome that were significantly associated with phenotypes such as potential serovar specific genes including capsule genes, and 48 putative virulence factors that were significantly different between the clinical and non-clinical isolates. We also show that the presence of many previously suggested virulence factors is not an appropriate marker of virulence. Conclusions: These genes will inform the generation of new molecular diagnostics and vaccines, and refinement of existing typing schemes and show the importance of the accessory genome of a diverse species when investigating the relationship between genotypes and phenotypes. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
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50. Antibiotic Sensitivity Profiles Do Not Reliably Distinguish Relapsing or Persisting Infections from Reinfections in Cats with Chronic Renal Failure and Multiple Diagnoses of Escherichia coli Urinary Tract Infection
- Author
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Freitag, Thurid, primary, Squires, Richard A., additional, Schmid, Jan, additional, Elliott, Jonathan, additional, and Rycroft, Andrew N., additional
- Published
- 2006
- Full Text
- View/download PDF
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