Genotyping of German and Austrian Taylorella equigenitalis isolates using repetitive extragenic palindromic (REP) PCR and pulsed-field gel electrophoresis (PFGE).
Abstract: A total of 124 Taylorella (T.) equigenitalis and five T. asinigenitalis field isolates collected between 2002 and 2014 were available for genotyping using REP- (repetitive extragenic palindromic) PCR and PFGE (pulsed-field gel electrophoresis). The study comprised 79 T. equigenitalis field isolates originating from ten defined breeds of German horses and revealed a spectrum of five REP (rep-E1-E4, rep-E3a) and 15 PFGE (TE-A1-A9, TE-B1-B3, TE-C, TE-E1, and TE-E2) genotypes. T. equigenitalis field isolates (n=40) obtained from Austrian Lipizzaner horses were differentiated into three REP (rep-E1, rep-E3a, and rep-E4) and three PFGE genotypes (TE-A2, TE-A5, and TE-D); those isolated from four Austrian Trotters belonged to the REP/PFGE genotype rep-E2/TE-A1. Interestingly, a T. equigenitalis isolate recovered from a Holsteiner stallion living in South Africa revealed the REP/PFGE genotype rep-E1/TE-A5 which was otherwise exclusively present in the majority of Austrian Lipizzaner horses in our study. The type strain included in this study revealed the genotype REP/PFGE rep-E1/TE-F. Six strains of T. asinigenitalis including the type strain were separated into three REP (rep-A1-A3) and six PFGE genotypes (TA-A1, TA-A2, TA-A3, TA-B, TA-C, TA-D). Overall, the generated REP and PFGE genotypes showed a good correlation, whereas REP-PCR proved to be a suitable method for molecular epidemiological screening of T. equigenitalis and T. asinigenitalis isolates that should be differentiated in detail by genotyping using PFGE.
Copyright © 2016 Elsevier Ltd. All rights reserved.
Publication Date: 2016-10-04 PubMed ID: 27892857DOI: 10.1016/j.rvsc.2016.09.017Google Scholar: Lookup
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- Journal Article
Summary
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This study conducted genotyping on 129 field isolates of two Taylorella bacteria species, collected between 2002 and 2014, using two different methods: repetitive extragenic palindromic (REP) PCR and pulsed-field gel electrophoresis (PFGE). The study findings suggest that the REP-PCR process is a suitable method for screening these bacteria, which should then be further differentiated using PFGE.
Types of Taylorella Studied
- The research focused on two species of Taylorella: T. equigenitalis and T.asinigenitalis.
- These bacteria have been isolated from a number of different breeds of horses in Germany and Austria, as well as from a stallion living in South Africa.
- The bacterial isolates studied were collected between the years 2002 and 2014.
Genotyping Methods Used
- The researchers used two different methods to perform genotyping on the bacterial isolates: REP-PCR and PFGE.
- REP-PCR method is based on examining the repeating sequences in the DNA. It was used to give a preliminary sorting of the bacterial isolates.
- PFGE method is a technique that allows for the separation and analysis of large DNA molecules by applying an electric field that periodically changes direction. It differentiates bacterial strains based on their genomic DNA profiles.
Results and Findings
- The study found a variety of genotypes among the bacterial isolates, with five REP and fifteen PFGE genotypes discovered among the T. equigenitalis bacteria alone.
- In the case of T. asinigenitalis, three REP and six PFGE genotypes were identified.
- The findings suggest that the REP-PCR method is a suitable and reliable method for preliminary screening of these bacteria, with PFGE used for further detailed differentiation.
- One T. equigenitalis isolate from a South African stallion had the same genotype as the majority of the Austrian Lipizzaner horses in the study.
Cite This Article
APA
Sting R, Seeh C, Mauder N, Maurer M, Loncaric I, Stessl B, Kopp P, Banzhaf K, Martin B, Melzer F, Raßbach A, Spergser J.
(2016).
Genotyping of German and Austrian Taylorella equigenitalis isolates using repetitive extragenic palindromic (REP) PCR and pulsed-field gel electrophoresis (PFGE).
Res Vet Sci, 109, 101-106.
https://doi.org/10.1016/j.rvsc.2016.09.017 Publication
Researcher Affiliations
- Chemisches und Veterinäruntersuchungsamt Stuttgart (Chemical and Veterinary Investigations Office Stuttgart), Schaflandstr. 3/3, Fellbach D-70736, Germany. Electronic address: reinhard.sting@cvuas.bwl.de.
- Pferdegesundheitsdienst Stuttgart, Tierseuchenkasse Baden-Württemberg (Animal Health Service Stuttgart, Animal Diseases Fund Baden-Wuerttemberg), Schaflandstr. 3/3, Fellbach D-70736, Germany.
- Chemisches und Veterinäruntersuchungsamt Stuttgart (Chemical and Veterinary Investigations Office Stuttgart), Schaflandstr. 3/3, Fellbach D-70736, Germany.
- Institut für Mikrobiologie, Veterinärmedizinische Universität Wien (Institute of Microbiology, University of Veterinary Medicine Vienna), Veterinärplatz 1, A-1210 Vienna, Austria.
- Institut für Mikrobiologie, Veterinärmedizinische Universität Wien (Institute of Microbiology, University of Veterinary Medicine Vienna), Veterinärplatz 1, A-1210 Vienna, Austria.
- Institut für Milchhygiene, Veterinärmedizinische Universität Wien (Institute for Milk Hygiene, Milk Technology and Food Science, University of Veterinary Medicine Vienna), Veterinärplatz 1, A-1210 Vienna, Austria.
- IDEXX Vet·Med·Labor, Mörikestr. 28/3, D-71636, Ludwigsburg, Germany.
- Pferdegesundheitsdienst Aulendorf, Tierseuchenkasse Baden-Württemberg (Animal Health Service Aulendorf, Animal Diseases Fund Baden-Wuerttemberg), Talstr. 17, D-88326, Aulendorf, Germany.
- Staatliches Tierärztliches Untersuchungsamt Aulendorf - Diagnostikzentrum (Aulendorf State Veterinary Diagnostic Centre), Löwenbreitestr. 18/20, D-88326, Aulendorf, Germany.
- Friedrich-Loeffler-Institut, Institut für bakterielle Infektionen und Zoonosen, Nationales Referenzlabor für Kontagiöse Equine Metritis (Institute of Bacterial Infections and Zoonoses, National Reference Laboratory for Contagious Equine Metritis), Naumburger Str. 96 a, D-07743, Jena, Germany.
- Friedrich-Loeffler-Institut, Institut für bakterielle Infektionen und Zoonosen, Nationales Referenzlabor für Kontagiöse Equine Metritis (Institute of Bacterial Infections and Zoonoses, National Reference Laboratory for Contagious Equine Metritis), Naumburger Str. 96 a, D-07743, Jena, Germany.
- Institut für Mikrobiologie, Veterinärmedizinische Universität Wien (Institute of Microbiology, University of Veterinary Medicine Vienna), Veterinärplatz 1, A-1210 Vienna, Austria.
MeSH Terms
- Animals
- Austria
- Electrophoresis, Gel, Pulsed-Field / veterinary
- Female
- Genotype
- Germany
- Gram-Negative Bacterial Infections / genetics
- Gram-Negative Bacterial Infections / microbiology
- Gram-Negative Bacterial Infections / veterinary
- Horse Diseases / genetics
- Horse Diseases / microbiology
- Horses
- Inverted Repeat Sequences
- Male
- Polymerase Chain Reaction / veterinary
- Taylorella equigenitalis / genetics
Citations
This article has been cited 3 times.- May CE, Guthrie AJ, Schulman ML. Direct culture-independent sequence typing of Taylorella equigenitalis obtained from genital swabs and frozen semen samples from South African horses. J Vet Diagn Invest 2019 Sep;31(5):792-794.
- Melzer F, Raßbach A, Köenig-Mozes A, Elschner MC, Tomaso H, Busch A. Draft Genome Sequence of Taylorella equigenitalis Strain 210217RC10635, Isolated from a Pony Stallion in Germany. Microbiol Resour Announc 2018 Sep;7(12).
- Grabatin M, Fux R, Zablotski Y, Goehring LS, Witte TS. Taylorella equigenitalis in Icelandic intact males compared with other horse breeds using natural cover. Equine Vet J 2025 Mar;57(2):441-448.
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