Sequencing-Based Genotyping of Pakistani Burkholderia mallei Strains: A Useful Way for Investigating Glanders Outbreaks.
Abstract: Burkholderia (B.) mallei is a host-adapted equine pathogen that causes glanders, a re-emerging zoonotic disease, which is endemic in Pakistan and other developing countries and seriously impacts the global equine movement. Due to globalization, the geographical restriction of diseases vanishes and the lack of awareness of and experience with eradicated diseases in industrialized countries also promotes the re-introduction of infections in these regions. Owing to the high equine population, the Pakistani province Punjab is a potential hotspot where several glanders outbreaks have been seen over last two decades. For determining the genomic diversity of B. mallei in this and other equine-populated prefectures, the genomes of 19 B. mallei strains isolated between 1999 and 2020 in different locations were sequenced and their genotypes were determined. Particularly, for genetically highly homogenous pathogens like B. mallei genotyping techniques require a high discriminatory power for enabling differentiation on the strain level. Thus, core-genome single nucleotide polymorphism (cgSNP) analysis was applied for distinguishing the highly similar strains. Furthermore, a whole-genome sequence-based core genome multi locus sequence typing (cgMLST) scheme, specific to B. mallei, was developed and additionally applied to the data. It was found that B. mallei genotypes in Pakistan persisted over time and space and genotype clusters preferred connection with a time point rather than the place of isolation, probably due to frequent equine movement, which promotes the spread of glanders. The cgMLST approach proved to work in accord with SNP typing and may help to investigate future glanders outbreaks.
Publication Date: 2022-05-24 PubMed ID: 35745468PubMed Central: PMC9227068DOI: 10.3390/pathogens11060614Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research conducted a genomic study on Burkholderia mallei strains, a bacteria causing a re-emerging zoonotic disease called glanders, from Pakistan. To do so, it used genotyping techniques on samples collected over two decades. It found that the strains of this bacteria in Pakistan have persisted historically and geographically, and that the risk of glanders outbreaks is tied more closely to timing than location.
Understanding the Research Context
- The research revolves around the bacterium Burkholderia mallei – the pathogen that causes glanders, a zoonotic disease which affects equines (e.g. horses) but can also infect humans. The bacterium has become a serious concern in Pakistan and other developing countries owing to its re-emerging nature and the substantial equine population, which promotes the spread of the disease.
- Historically, globalization has lifted geographical boundaries, causing diseases to spread more easily across countries. This makes the study of such diseases vital to prevent reintroduction into previously unaffected or eradicated areas.
Methodology and Key Findings
- The genomes of 19 different Burkholderia mallei strains, isolated between 1999 and 2020, were sequenced to determine their genotype. This helps to understand the genetic diversity of the strains within the region.
- The researchers applied a specific technique known as core-genome single nucleotide polymorphism (cgSNP) analysis. This is particularly useful in differentiating highly similar strains of homogenous pathogens like Burkholderia mallei, as it offers high discriminatory power.
- The team then developed and applied a whole-genome sequence-based core genome multi locus sequence typing (cgMLST) scheme specific to Burkholderia mallei. This provided additional differentiation of the collected strains.
- The results suggested that the genotypes of Burkholderia mallei in Pakistan have persisted over time and space. This persistence indicates selection pressures may be in play, supporting survival and proliferation of certain genetic types versus others.
- Interestingly, the study found the genetic clusters formed by the bacterium were more strongly associated with specific time points than with their place of isolation. This supports the concept that the frequent movement of equines, rather than location, largely promotes the spread of glanders.
Implications of the Study
- The researchers highlighted the effectiveness of the cgMLST approach in conjunction with SNP typing for distinguishing Burkholderia mallei strains. Such techniques can provide timely and accurate genotyping of the bacterium, which is invaluable for tracking and investigating future outbreaks of glanders.
- This understanding of strain diversity and spread may allow better control and prevention measures in regions with high equine population density and frequent equine movement.
Cite This Article
APA
Brangsch H, Saqib M, Sial AUR, Melzer F, Linde J, Elschner MC.
(2022).
Sequencing-Based Genotyping of Pakistani Burkholderia mallei Strains: A Useful Way for Investigating Glanders Outbreaks.
Pathogens, 11(6).
https://doi.org/10.3390/pathogens11060614 Publication
Researcher Affiliations
- Institute for Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institute, Naumburger Str. 96a, 07743 Jena, Thuringia, Germany.
- Veterinary Preventive Medicine and Public Health Laboratory, Department of Clinical Medicine and Surgery, University of Agriculture, Faisalabad 38000, Pakistan.
- Department of Clinical Studies, Faculty of Veterinary and Animal Science, Pir Mehr Ali Shah, Arid Agriculture University Rawalpindi, Rawalpindi 46000, Pakistan.
- Institute for Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institute, Naumburger Str. 96a, 07743 Jena, Thuringia, Germany.
- Institute for Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institute, Naumburger Str. 96a, 07743 Jena, Thuringia, Germany.
- Institute for Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institute, Naumburger Str. 96a, 07743 Jena, Thuringia, Germany.
Conflict of Interest Statement
The authors declare no conflict of interest.
References
This article includes 47 references
- Hornstra H, Pearson T, Georgia S, Liguori A, Dale J, Price E, O’Neill M, Deshazer D, Muhammad G, Saqib M. Molecular epidemiology of glanders, Pakistan.. Emerg. Infect. Dis. 2009;15:2036–2039.
- Timoney PJ. Infectious diseases and international movement of horses.. Equine Infectious Diseases 2014;pp. 544–551.e541.
- Fonseca-Rodriguez O, Pinheiro Junior JW, Mota RA. Spatiotemporal analysis of glanders in Brazil.. J. Equine Vet. Sci. 2019;78:14–19.
- Gonzalez-Medina S, Toth B, Mawhinney I. Surveillance focus: Glanders.. Vet. Rec. 2015;177:68–69.
- Van Zandt KE, Greer MT, Gelhaus HC. Glanders: An overview of infection in humans.. Orphanet J. Rare Dis. 2013;8:131.
- Srinivasan A, Kraus CN, DeShazer D, Becker PM, Dick JD, Spacek L, Bartlett JG, Byrne WR, Thomas DL. Glanders in a military research microbiologist.. N. Engl. J. Med. 2001;345:256–258.
- Saqib M, Muhammad G, Naureen A, Hussain MH, Asi MN, Mansoor MK, Toufeer M, Khan I, Neubauer H, Sprague LD. Effectiveness of an antimicrobial treatment scheme in a confined glanders outbreak.. BMC Vet. Res. 2012;8:214.
- Wang G, Zarodkiewicz P, Valvano MA. Current advances in Burkholderia vaccines development.. Cells 2020;9:2671.
- Arun S, Neubauer H, Gurel A, Ayyildiz G, Kuscu B, Yesildere T, Meyer H, Hermanns W. Equine glanders in Turkey.. Vet. Rec. 1999;144:255–258.
- Kettle AN, Wernery U. Glanders and the risk for its introduction through the international movement of horses.. Equine Vet. J. 2016;48:654–658.
- Elschner MC, Klaus CU, Liebler-Tenorio E, Schmoock G, Wohlsein P, Tinschmann O, Lange E, Kaden V, Klopfleisch R, Melzer F. Burkholderia mallei infection in a horse imported from Brazil.. Equine Vet. Educ. 2009;21:147–150.
- Khan I, Wieler LH, Melzer F, Elschner MC, Muhammad G, Ali S, Sprague LD, Neubauer H, Saqib M. Glanders in animals: A review on epidemiology, clinical presentation, diagnosis and countermeasures.. Transbound. Emerg. Dis. 2013;60:204–221.
- . Glanders and farcy in India.. Vet. J. 1877;5:424–427.
- Sial AUR, Saqib M, Muhammad G, Sajid MS. Seroprevalence and risk factors of equine glanders in selected districts of Khyber Pakhtunkhwa (KPK). Pak. Vet. J. 2020;40:504–508.
- Khan I, Ali S, Gwida M, Elschner M, Ijaz A, Neubauer H. Prevalence of Burkholderia mallei in equids of remount depot, Sargodha, Pakistan.. Pak. J. Zool. 2013;45:1751–1756.
- Khan I, Wieler LH, Butt MA, Elschner MC, Cheema AH, Sprague LD, Neubauer H. On the current situation of glanders in various districts of the Pakistani Punjab.. J. Equine Vet. Sci. 2012;32:783–787.
- Malik P, Singha H, Khurana S, Kumar R, Kumar S, Raut A, Riyesh T, Vaid RK, Virmani N, Singh B. Emergence and re-emergence of glanders in India: A description of outbreaks from 2006 to 2011.. Vet. Ital. 2012;48:167–178.
- Verma AK, Saminathan M, Tiwari R, Dhama K, Singh SV. Glanders—A re-emerging zoonotic disease: A review.. J. Biol. Sci. 2014;14:38–51.
- Muhammad G, Khan MZ, Athar M. Clinico-microbiological and therapeutic aspects of glanders in equines.. J. Equine Sci. 1998;9:93–96.
- Losada L, Ronning CM, DeShazer D, Woods D, Fedorova N, Kim HS, Shabalina SA, Pearson TR, Brinkac L, Tan P. Continuing evolution of Burkholderia mallei through genome reduction and large-scale rearrangements.. Genome Biol. Evol. 2010;2:102–116.
- Godoy D, Randle G, Simpson AJ, Aanensen DM, Pitt TL, Kinoshita R, Spratt BG. Multilocus sequence typing and evolutionary relationships among the causative agents of melioidosis and glanders, Burkholderia pseudomallei and Burkholderia mallei.. J. Clin. Microbiol. 2003;41:2068–2079.
- U’Ren JM, Schupp JM, Pearson T, Hornstra H, Friedman CL, Smith KL, Daugherty RR, Rhoton SD, Leadem B, Georgia S. Tandem repeat regions within the Burkholderia pseudomallei genome and their application for high resolution genotyping.. BMC Microbiol. 2007;7:23.
- Keim P, Van Ert MN, Pearson T, Vogler AJ, Huynh LY, Wagner DM. Anthrax molecular epidemiology and forensics: Using the appropriate marker for different evolutionary scales.. Infect. Genet. Evol. 2004;4:205–213.
- Girault G, Wattiau P, Saqib M, Martin B, Vorimore F, Singha H, Engelsma M, Roest HJ, Spicic S, Grunow R. High-resolution melting PCR analysis for rapid genotyping of Burkholderia mallei.. Infect. Genet. Evol. 2018;63:1–4.
- Laroucau K, Lucia de Assis Santana V, Girault G, Martin B, Miranda da Silveira PP, Brasil Machado M, Joseph M, Wernery R, Wernery U, Zientara S. First molecular characterisation of a Brazilian Burkholderia mallei strain isolated from a mule in 2016.. Infect. Genet. Evol. 2018;57:117–120.
- Scholz HC, Pearson T, Hornstra H, Projahn M, Terzioglu R, Wernery R, Georgi E, Riehm JM, Wagner DM, Keim PS. Genotyping of Burkholderia mallei from an outbreak of glanders in Bahrain suggests multiple introduction events.. PLoS Negl. Trop. Dis. 2014;8:e3195.
- De Been M, Pinholt M, Top J, Bletz S, Mellmann A, van Schaik W, Brouwer E, Rogers M, Kraat Y, Bonten M. Core genome multilocus sequence typing scheme for high-resolution typing of Enterococcus faecium.. J. Clin. Microbiol. 2015;53:3788–3797.
- Pearce ME, Alikhan NF, Dallman TJ, Zhou Z, Grant K, Maiden MCJ. Comparative analysis of core genome MLST and SNP typing within a European Salmonella serovar Enteritidis outbreak.. Int. J. Food Microbiol. 2018;274:1–11.
- Higgins PG, Prior K, Harmsen D, Seifert H. Development and evaluation of a core genome multilocus typing scheme for whole-genome sequence-based typing of Acinetobacter baumannii.. PLoS ONE. 2017;12:e0179228.
- Abdel-Glil MY, Thomas P, Linde J, Jolley KA, Harmsen D, Wieler LH, Neubauer H, Seyboldt C. Establishment of a publicly available core genome multilocus sequence typing scheme for Clostridium perfringens.. Microbiol. Spectr. 2021;9:e0053321.
- Ruppitsch W, Pietzka A, Prior K, Bletz S, Fernandez HL, Allerberger F, Harmsen D, Mellmann A. Defining and evaluating a core genome multilocus sequence typing scheme for whole-genome sequence-based typing of Listeria monocytogenes.. J. Clin. Microbiol. 2015;53:2869–2876.
- Lichtenegger S, Trinh Trung T, Assig K, Prior K, Harmsen D, Pesl J, Zauner A, Lipp M, Que Tram A, Mutsam B. Development and validation of a Burkholderia pseudomallei core genome multilocus sequence typing scheme to facilitate molecular surveillance.. J. Clin. Microbiol. 2021;59:e00093-21.
- Sahl JW, Vazquez AJ, Hall CM, Busch JD, Tuanyok A, Mayo M, Schupp JM, Lummis M, Pearson T, Shippy K. The effects of signal erosion and core genome reduction on the identification of diagnostic markers.. MBio. 2016;7:e00846-16.
- Ghori MT, Khan MS, Khan JA, Rabbani M, Shabbir MZ, Chaudhry HR, Ali MA, Muhammad J, Elschner MC, Jayarao BM. Seroprevalence and risk factors of glanders in working equines—Findings of a cross-sectional study in Punjab province of Pakistan.. Acta Trop. 2017;176:134–139.
- Romero CM, DeShazer D, Feldblyum T, Ravel J, Woods D, Kim HS, Yu Y, Ronning CM, Nierman WC. Genome sequence alterations detected upon passage of Burkholderia mallei ATCC 23344 in culture and in mammalian hosts.. BMC Genom. 2006;7:228.
- Laroucau K, Aaziz R, Vorimore F, Varghese K, Deshayes T, Bertin C, Delannoy S, Sami AM, Al Batel M, El Shorbagy M. A genetic variant of Burkholderia mallei detected in Kuwait: Consequences for the PCR diagnosis of glanders.. Transbound. Emerg. Dis. 2021;68:960–963.
- Ulrich RL, Ulrich MP, Schell MA, Kim HS, DeShazer D. Development of a polymerase chain reaction assay for the specific identification of Burkholderia mallei and differentiation from Burkholderia pseudomallei and other closely related Burkholderiaceae.. Diagn. Microbiol. Infect. Dis. 2006;55:37–45.
- Moore D. Preparation and analysis of DNA.. Current Protocols in Molecular Biology 1995.
- De Coster W, D’Hert S, Schultz DT, Cruts M, Van Broeckhoven C. NanoPack: Visualizing and processing long-read sequencing data.. Bioinformatics 2018;34:2666–2669.
- Wick RR, Judd LM, Gorrie CL, Holt KE. Unicycler: Resolving bacterial genome assemblies from short and long sequencing reads.. PLoS Comput. Biol. 2017;13:e1005595.
- Wood DE, Lu J, Langmead B. Improved metagenomic analysis with Kraken 2.. Genome Biol. 2019;20:257.
- Gurevich A, Saveliev N, Vyahhi N, Tesler G. QUAST: Quality assessment tool for genome assemblies.. Bioinformatics 2013;29:1072–1075.
- Seemann T. Prokka: Rapid prokaryotic genome annotation.. Bioinformatics 2014;30:2068–2069.
- Jünemann S, Sedlazeck FJ, Prior K, Albersmeier A, John U, Kalinowski J, Mellmann A, Goesmann A, von Haeseler A, Stoye J. Updating benchtop sequencing performance comparison.. Nat. Biotechnol. 2013;31:294–296.
- Stamatakis A. RAxML version 8: A tool for phylogenetic analysis and post-analysis of large phylogenies.. Bioinformatics 2014;30:1312–1313.
- Treangen TJ, Ondov BD, Koren S, Phillippy AM. The Harvest suite for rapid core-genome alignment and visualization of thousands of intraspecific microbial genomes.. Genome Biol. 2014;15:524.
- Huson DH, Scornavacca C. Dendroscope 3: An interactive tool for rooted phylogenetic trees and networks.. Syst. Biol. 2012;61:1061–1067.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists