Sequence variation of the SeM gene of Streptococcus equi allows discrimination of the source of strangles outbreaks.
Abstract: Improved understanding of the epidemiology of Streptococcus equi transmission requires sensitive and portable subtyping methods that can rationally discriminate between strains. S. equi is highly homogeneous and cannot be distinguished by multilocus enzyme electrophoretic or multilocus sequence-typing methods that utilize housekeeping genes. However, on sequence analysis of the N-terminal region of the SeM genes of 60 S. equi isolates from 27 strangles outbreaks, we identified 21 DNA codon changes. These resulted in the nonsynonymous substitution of 18 amino acids and allowed the assignment of S. equi strains to 15 distinct subtypes. Our data suggest the presence of multiple epitopes across this region that are subjected to selective immune pressure (nonsynonymous-synonymous substitution rate [d(N)/d(S)] ratio = 3.054), particularly during the establishment of long-term S. equi infection. We further report the application of SeM gene subtyping as a method to investigate potential cases of disease related to administration of a live attenuated S. equi vaccine. SeM gene subtyping successfully differentiated between the vaccine strain and field strains of S. equi responsible for concurrent disease. These results were confirmed by the development and application of a PCR diagnostic test, which identifies the aroA partial gene deletion present in the Equilis StrepE vaccine strain. Although the vaccine strain was found to be responsible for injection site lesions, all seven outbreaks of strangles investigated in recently vaccinated horses were found to be due to concurrent infection with wild-type S. equi and not due to reversion of the vaccine strain.
Publication Date: 2006-02-04 PubMed ID: 16455902PubMed Central: PMC1392674DOI: 10.1128/JCM.44.2.480-486.2006Google Scholar: Lookup
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Summary
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The study focuses on identifying distinct strains of Streptococcus equi – the bacterium that causes strangles in horses – to determine the origin of disease outbreaks. Researchers observed 21 DNA codon changes in a specific gene, allowing them to discriminate between 15 different subtypes of the bacteria. The study also investigated potential disease cases related to a live vaccine, successfully differentiating the vaccine strain from wild strains. The results indicated that the vaccine strain could cause injection site reactions but was not responsible for disease outbreaks.
Identification of Streptococcus equi Strains
- The researchers wanted an improved method to identify strains of the bacterium Streptococcus equi, which causes the equine disease strangles. Understanding the exact strains involved in outbreaks can inform more effective treatment and prevention strategies.
- This bacterium is highly similar across strains, making it difficult to distinguish between different types using conventional methods. Hence, the researchers conducted a sequence analysis of the N-terminal region of the SeM genes of 60 S. equi isolates from 27 strangles outbreaks.
- Through this analysis, they found 21 DNA codon changes that resulted in the nonsynonymous substitution of 18 amino acids, allowing them to categorize the S. equi strains into 15 distinct subtypes. These variations are due to selective immune pressure on certain gene segments – a factor that contributes to the evolution and differentiation of bacterial strains.
Investigation of Live Vaccine-Related Disease Cases
- As part of the study, the researchers also investigated potential cases of disease related to the administration of a live attenuated S. equi vaccine, known as the Equilis StrepE vaccine.
- The researchers developed and employed a PCR diagnostic test that identifies a partial gene deletion in the vaccine strain. This enabled them to successfully differentiate the vaccine strain from field strains of S. equi responsible for concurrent disease.
- The findings showed that while the vaccine strain can be responsible for injection site lesions, it was not the cause of the seven strangles outbreaks investigated in the study among recently vaccinated horses. All outbreaks were due to concurrent infection with wild-type S. equi and not due to reversion of the vaccine strain.
Implications of the Research
- This research demonstrates the applicability and effectiveness of SeM gene subtyping as a method to identify distinct strains of S. equi and to track the source of disease outbreaks.
- The ability to differentiate between the vaccine strain and wild strains can counter any mistaken attribution of illness to the vaccine, maintaining confidence in its use and supplying crucial information for future vaccine development.
- Finally, this detailed understanding of S. equi strains can contribute to improved treatment and prevention strategies for strangles.
Cite This Article
APA
Kelly C, Bugg M, Robinson C, Mitchell Z, Davis-Poynter N, Newton JR, Jolley KA, Maiden MC, Waller AS.
(2006).
Sequence variation of the SeM gene of Streptococcus equi allows discrimination of the source of strangles outbreaks.
J Clin Microbiol, 44(2), 480-486.
https://doi.org/10.1128/JCM.44.2.480-486.2006 Publication
Researcher Affiliations
- Centre for Preventive Medicine, Animal Health Trust, Lanwades Park, Kentford, Newmarket, Suffolk CB8 7UU, United Kingdom.
MeSH Terms
- Amino Acid Sequence
- Animals
- Antigens, Bacterial / chemistry
- Antigens, Bacterial / genetics
- Bacterial Proteins / chemistry
- Bacterial Proteins / genetics
- Disease Outbreaks
- Gene Deletion
- Genetic Variation
- Horse Diseases / epidemiology
- Horse Diseases / microbiology
- Horse Diseases / prevention & control
- Horses
- Molecular Sequence Data
- Polymerase Chain Reaction / methods
- Streptococcal Infections / epidemiology
- Streptococcal Infections / microbiology
- Streptococcal Infections / prevention & control
- Streptococcal Infections / veterinary
- Streptococcal Vaccines / administration & dosage
- Streptococcal Vaccines / genetics
- Streptococcus equi / classification
- Streptococcus equi / genetics
- Streptococcus equi / immunology
- Streptococcus equi / isolation & purification
- Vaccination / veterinary
- Vaccines, Attenuated / administration & dosage
- Vaccines, Attenuated / genetics
References
This article includes 36 references
- Al-Ghamdi GM, Kapur V, Ames TR, Timoney JF, Love DN, Mellencamp MA. Use of repetitive sequence-based polymerase chain reaction for molecular epidemiologic analysis of Streptococcus equi subspecies equi.. Am J Vet Res 2000 Jun;61(6):699-705.
- Boschwitz JS, Timoney JF. Inhibition of C3 deposition on Streptococcus equi subsp. equi by M protein: a mechanism for survival in equine blood.. Infect Immun 1994 Aug;62(8):3515-20.
- Chanter N, Talbot NC, Newton JR, Hewson D, Verheyen K. Streptococcus equi with truncated M-proteins isolated from outwardly healthy horses.. Microbiology (Reading) 2000 Jun;146 ( Pt 6):1361-1369.
- Galán JE, Timoney JF. Molecular analysis of the M protein of Streptococcus equi and cloning and expression of the M protein gene in Escherichia coli.. Infect Immun 1987 Dec;55(12):3181-7.
- Galán JE, Timoney JF. Immunologic and genetic comparison of Streptococcus equi isolates from the United States and Europe.. J Clin Microbiol 1988 Jun;26(6):1142-6.
- Harrington DJ, Sutcliffe IC, Chanter N. The molecular basis of Streptococcus equi infection and disease.. Microbes Infect 2002 Apr;4(4):501-10.
- Hartford, O. M., T. J. Foster, and A. A. C. Jacobs. April 1999. United States Patent 5,895,654.
- Jacobs AA, Goovaerts D, Nuijten PJ, Theelen RP, Hartford OM, Foster TJ. Investigations towards an efficacious and safe strangles vaccine: submucosal vaccination with a live attenuated Streptococcus equi.. Vet Rec 2000 Nov 11;147(20):563-7.
- Jorm LR, Love DN, Bailey GD, McKay GM, Briscoe DA. Genetic structure of populations of beta-haemolytic Lancefield group C streptococci from horses and their association with disease.. Res Vet Sci 1994 Nov;57(3):292-9.
- Karlström A, Jacobsson K, Flock M, Flock JI, Guss B. Identification of a novel collagen-like protein, SclC, in Streptococcus equi using signal sequence phage display.. Vet Microbiol 2004 Dec 9;104(3-4):179-88.
- Kumar S, Tamura K, Nei M. MEGA: Molecular Evolutionary Genetics Analysis software for microcomputers.. Comput Appl Biosci 1994 Apr;10(2):189-91.
- Lannergård J, Frykberg L, Guss B. CNE, a collagen-binding protein of Streptococcus equi.. FEMS Microbiol Lett 2003 May 16;222(1):69-74.
- Lindmark H, Guss B. SFS, a novel fibronectin-binding protein from Streptococcus equi, inhibits the binding between fibronectin and collagen.. Infect Immun 1999 May;67(5):2383-8.
- Lindmark H, Jonsson P, Engvall E, Guss B. Pulsed-field gel electrophoresis and distribution of the genes zag and fnz in isolates of Streptococcus equi.. Res Vet Sci 1999 Apr;66(2):93-9.
- Meehan M, Nowlan P, Owen P. Affinity purification and characterization of a fibrinogen-binding protein complex which protects mice against lethal challenge with Streptococcus equi subsp. equi.. Microbiology (Reading) 1998 Apr;144 ( Pt 4):993-1003.
- Meehan M, Muldowney DA, Watkins NJ, Owen P. Localization and characterization of the ligand-binding domain of the fibrinogen-binding protein (FgBP) of Streptococcus equi subsp. equi.. Microbiology (Reading) 2000 May;146 ( Pt 5):1187-1194.
- Meehan M, Lynagh Y, Woods C, Owen P. The fibrinogen-binding protein (FgBP) of Streptococcus equi subsp. equi additionally binds IgG and contributes to virulence in a mouse model.. Microbiology (Reading) 2001 Dec;147(Pt 12):3311-22.
- Meinersmann RJ, Phillips RW, Wiedmann M, Berrang ME. Multilocus sequence typing of Listeria monocytogenes by use of hypervariable genes reveals clonal and recombination histories of three lineages.. Appl Environ Microbiol 2004 Apr;70(4):2193-203.
- Nei M, Gojobori T. Simple methods for estimating the numbers of synonymous and nonsynonymous nucleotide substitutions.. Mol Biol Evol 1986 Sep;3(5):418-26.
- Newton JR, Wood JL, Dunn KA, DeBrauwere MN, Chanter N. Naturally occurring persistent and asymptomatic infection of the guttural pouches of horses with Streptococcus equi.. Vet Rec 1997 Jan 25;140(4):84-90.
- Newton JR, Verheyen K, Talbot NC, Timoney JF, Wood JL, Lakhani KH, Chanter N. Control of strangles outbreaks by isolation of guttural pouch carriers identified using PCR and culture of Streptococcus equi.. Equine Vet J 2000 Nov;32(6):515-26.
- Newton R, Waller A, King A. Investigation of suspected adverse reactions following strangles vaccination in horses.. Vet Rec 2005 Feb 26;156(9):291-2.
- Page RD. TreeView: an application to display phylogenetic trees on personal computers.. Comput Appl Biosci 1996 Aug;12(4):357-8.
- Posada D, Crandall KA. MODELTEST: testing the model of DNA substitution.. Bioinformatics 1998;14(9):817-8.
- Reid SD, Herbelin CJ, Bumbaugh AC, Selander RK, Whittam TS. Parallel evolution of virulence in pathogenic Escherichia coli.. Nature 2000 Jul 6;406(6791):64-7.
- Sheoran AS, Artiushin S, Timoney JF. Nasal mucosal immunogenicity for the horse of a SeM peptide of Streptococcus equi genetically coupled to cholera toxin.. Vaccine 2002 Feb 22;20(11-12):1653-9.
- Swofford, D. L. 2003. PAUP*. Phylogenetic analysis using parsimony (*and 15 other methods), version 4. Sinauer Associates, Sunderland, Mass.
- Takai S, Anzai T, Yashiro H, Ishii C, Tsubaki S, Wada R, Timoney JF. Detection of DNA restriction fragment polymorphisms in Streptococcus equi.. Vet Rec 2000 Feb 5;146(6):159-61.
- Timoney JF, Eggers D. Serum bactericidal responses to Streptococcus equi of horses following infection or vaccination.. Equine Vet J 1985 Jul;17(4):306-10.
- Timoney JF. Strangles.. Vet Clin North Am Equine Pract 1993 Aug;9(2):365-74.
- Timoney JF, Artiushin SC, Boschwitz JS. Comparison of the sequences and functions of Streptococcus equi M-like proteins SeM and SzPSe.. Infect Immun 1997 Sep;65(9):3600-5.
- Timoney JF, Artiushin SC, Wang J. Regions of the protective M-like protein of Streptococcus equi recognized by serum and mucosal antibodies of convalescent horses. Equine infectious diseases VIII 1999;p. 88-94.
- van Loo IH, Heuvelman KJ, King AJ, Mooi FR. Multilocus sequence typing of Bordetella pertussis based on surface protein genes.. J Clin Microbiol 2002 Jun;40(6):1994-2001.
- Verheyen K, Newton JR, Talbot NC, de Brauwere MN, Chanter N. Elimination of guttural pouch infection and inflammation in asymptomatic carriers of Streptococcus equi.. Equine Vet J 2000 Nov;32(6):527-32.
- Zadoks RN, Schukken YH, Wiedmann M. Multilocus sequence typing of Streptococcus uberis provides sensitive and epidemiologically relevant subtype information and reveals positive selection in the virulence gene pauA.. J Clin Microbiol 2005 May;43(5):2407-17.
- Zhang W, Jayarao BM, Knabel SJ. Multi-virulence-locus sequence typing of Listeria monocytogenes.. Appl Environ Microbiol 2004 Feb;70(2):913-20.
Citations
This article has been cited 23 times.- Knox A, Zerna G, Beddoe T. Current and Future Advances in the Detection and Surveillance of Biosecurity-Relevant Equine Bacterial Diseases Using Loop-Mediated Isothermal Amplification (LAMP).. Animals (Basel) 2023 Aug 18;13(16).
- Rotinsulu DA, Ewers C, Kerner K, Amrozi A, Soejoedono RD, Semmler T, Bauerfeind R. Molecular Features and Antimicrobial Susceptibilities of Streptococcus equi ssp. equi Isolates from Strangles Cases in Indonesia.. Vet Sci 2023 Jan 10;10(1).
- Morris ERA, Wu J, Bordin AI, Lawhon SD, Cohen ND. Differences in the Accessory Genomes and Methylomes of Strains of Streptococcus equi subsp. equi and of Streptococcus equi subsp. zooepidemicus Obtained from the Respiratory Tract of Horses from Texas.. Microbiol Spectr 2022 Feb 23;10(1):e0076421.
- Frosth S, Morris ERA, Wilson H, Frykberg L, Jacobsson K, Parkhill J, Flock JI, Wood T, Guss B, Aanensen DM, Boyle AG, Riihimäki M, Cohen ND, Waller AS. Conservation of vaccine antigen sequences encoded by sequenced strains of Streptococcus equi subsp. equi.. Equine Vet J 2023 Jan;55(1):92-101.
- Kerstens J, Durmus B, Lambrecht S, Baar I, Ieven MM, Van Der Zijden T, Parizel PM, Menovsky T, Lammens MMY, Jorens PG. Meningoencephalitis with Streptococcus equi Subspecies equi Leading to a Dural Arteriovenous Fistula.. Case Rep Neurol Med 2021;2021:9898364.
- Morris ERA, Boyle AG, Riihimäki M, Aspán A, Anis E, Hillhouse AE, Ivanov I, Bordin AI, Pringle J, Cohen ND. Differences in the genome, methylome, and transcriptome do not differentiate isolates of Streptococcus equi subsp. equi from horses with acute clinical signs from isolates of inapparent carriers.. PLoS One 2021;16(6):e0252804.
- Mitchell C, Steward KF, Charbonneau ARL, Walsh S, Wilson H, Timoney JF, Wernery U, Joseph M, Craig D, van Maanen K, Hoogkamer-van Gennep A, Leon A, Witkowski L, Rzewuska M, Stefańska I, Żychska M, van Loon G, Cursons R, Patty O, Acke E, Gilkerson JR, El-Hage C, Allen J, Bannai H, Kinoshita Y, Niwa H, Becú T, Pringle J, Guss B, Böse R, Abbott Y, Katz L, Leggett B, Buckley TC, Blum SE, Cruz López F, Fernández Ros A, Marotti Campi MC, Preziuso S, Robinson C, Newton JR, Schofield E, Brooke B, Boursnell M, de Brauwere N, Kirton R, Barton CK, Abudahab K, Taylor B, Yeats CA, Goater R, Aanensen DM, Harris SR, Parkhill J, Holden MTG, Waller AS. Globetrotting strangles: the unbridled national and international transmission of Streptococcus equi between horses.. Microb Genom 2021 Mar;7(3).
- Costa MO, Lage B. Streptococcus equi Subspecies zooepidemicus and Sudden Deaths in Swine, Canada.. Emerg Infect Dis 2020 Oct;26(10):2522-2524.
- Charbonneau ARL, Taylor E, Mitchell CJ, Robinson C, Cain AK, Leigh JA, Maskell DJ, Waller AS. Identification of genes required for the fitness of Streptococcus equi subsp. equi in whole equine blood and hydrogen peroxide.. Microb Genom 2020 Apr;6(4).
- D'Gama JD, Ma Z, Zhang H, Liu X, Fan H, Morris ERA, Cohen ND, Cywes-Bentley C, Pier GB, Waldor MK. A Conserved Streptococcal Virulence Regulator Controls the Expression of a Distinct Class of M-Like Proteins.. mBio 2019 Oct 22;10(5).
- Kasuya K, Tanaka N, Oshima F, Fujisawa N, Saito M, Tagami K, Niwa H, Sasai K. Genetic analysis of Streptococcus equi subsp. equi isolated from horses imported into Japan.. J Vet Med Sci 2019 Jun 28;81(6):924-927.
- Boyle AG, Timoney JF, Newton JR, Hines MT, Waller AS, Buchanan BR. Streptococcus equi Infections in Horses: Guidelines for Treatment, Control, and Prevention of Strangles-Revised Consensus Statement.. J Vet Intern Med 2018 Mar;32(2):633-647.
- Tirosh-Levy S, Blum SE, Steward KF, Waller AS, Steinman A. Streptococcus equi subspecies equi in horses in Israel: seroprevalence and strain types.. Vet Rec Open 2016;3(1):e000187.
- Harris SR, Robinson C, Steward KF, Webb KS, Paillot R, Parkhill J, Holden MT, Waller AS. Genome specialization and decay of the strangles pathogen, Streptococcus equi, is driven by persistent infection.. Genome Res 2015 Sep;25(9):1360-71.
- Hobo S, Niwa H, Anzai T, Jones JH. Changes in Serum Antibody Levels after Vaccination for Strangles and after Intranasal Challenge with Streptococcus equi subsp. equi in Horses.. J Equine Sci 2010;21(3):33-7.
- Moloney E, Kavanagh KS, Buckley TC, Cooney JC. Lineages of Streptococcus equi ssp. equi in the Irish equine industry.. Ir Vet J 2013;66(1):10.
- Webb K, Barker C, Harrison T, Heather Z, Steward KF, Robinson C, Newton JR, Waller AS. Detection of Streptococcus equi subspecies equi using a triplex qPCR assay.. Vet J 2013 Mar;195(3):300-4.
- Guss B, Flock M, Frykberg L, Waller AS, Robinson C, Smith KC, Flock JI. Getting to grips with strangles: an effective multi-component recombinant vaccine for the protection of horses from Streptococcus equi infection.. PLoS Pathog 2009 Sep;5(9):e1000584.
- Holden MT, Heather Z, Paillot R, Steward KF, Webb K, Ainslie F, Jourdan T, Bason NC, Holroyd NE, Mungall K, Quail MA, Sanders M, Simmonds M, Willey D, Brooks K, Aanensen DM, Spratt BG, Jolley KA, Maiden MC, Kehoe M, Chanter N, Bentley SD, Robinson C, Maskell DJ, Parkhill J, Waller AS. Genomic evidence for the evolution of Streptococcus equi: host restriction, increased virulence, and genetic exchange with human pathogens.. PLoS Pathog 2009 Mar;5(3):e1000346.
- Heather Z, Holden MT, Steward KF, Parkhill J, Song L, Challis GL, Robinson C, Davis-Poynter N, Waller AS. A novel streptococcal integrative conjugative element involved in iron acquisition.. Mol Microbiol 2008 Dec;70(5):1274-92.
- Meeusen EN, Walker J, Peters A, Pastoret PP, Jungersen G. Current status of veterinary vaccines.. Clin Microbiol Rev 2007 Jul;20(3):489-510, table of contents.
- Ladlow J, Scase T, Waller A. Canine strangles case reveals a new host susceptible to infection with Streptococcus equi.. J Clin Microbiol 2006 Jul;44(7):2664-5.
- Jolley KA, Maiden MC. AgdbNet - antigen sequence database software for bacterial typing.. BMC Bioinformatics 2006 Jun 21;7:314.
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