Whole-Genome Sequence of the
 Mycoplasma
 (
 Mesomycoplasma
 )
 hyorhinis
 DSM 25591 Type Strain

Käbisch, Lisa; Schink, Anne‐Kathrin; Hanke, Dennis; Semmler, Torsten; Kehrenberg, Corinna; Schwarz, Štefan

doi:10.1128/mra.00164-21

Cited by 4 publications

(7 citation statements)

References 15 publications

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“…Tetracyclines, macrolides, and pleuromutilins were of particular interest, since antimicrobial agents of these classes are commonly used to treat porcine respiratory infections caused by Mycoplasma ( ‘Mesomycoplasma’ ) spp. Erythromycin was included because several studies reported an intrinsic resistance to this 14-membered macrolide in M. hyopneumoniae as well as M. hyorhinis [ 11 , 12 , 13 , 14 ]. Lincosamides are also of interest for the treatment of porcine Mycoplasma ( ‘Mesomycoplasma’ ) spp.…”

Section: Resultsmentioning

confidence: 99%

Provisional Use of CLSI-Approved Quality Control Strains for Antimicrobial Susceptibility Testing of Mycoplasma (‘Mesomycoplasma’) hyorhinis

et al. 2021

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Antimicrobial susceptibility testing (AST) should be conducted in a standardized manner prior to the start of an antimicrobial treatment. For fastidious bacteria, such as porcine Mycoplasma (‘Mesomycoplasma’) spp., specifically M. hyorhinis, neither guidelines or standards for the performance of AST, nor quality control strains for the validation of AST results are approved by organizations like the Clinical and Laboratory Standards Institute (CLSI) or the European Committee on Antimicrobial Susceptibility Testing (EUCAST). The CLSI- and EUCAST-approved quality control strains Enterococcus faecalis ATCC 29212 and Staphylococcus aureus ATCC 29213 were chosen to validate AST by broth microdilution using modified Friis broth, developed as growth medium for porcine Mycoplasma (‘Mesomycoplasma’) spp. The antimicrobial agents doxycycline, enrofloxacin, erythromycin, florfenicol, gentamicin, marbofloxacin, tetracycline, tiamulin, tilmicosin, tulathromycin, and tylosin were examined using customized SensititreTM microtiter plates. Minimal inhibitory concentrations, determined after 24, 48, and 72 h, were mostly within the CLSI-approved quality control ranges for defined antimicrobial agents. We propose the use of the combination of E. faecalis ATCC 29212 and S. aureus ATCC 29213 as surrogate quality control strains for the validation of future AST results obtained for M. hyorhinis by broth microdilution using modified Friis broth.

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Section: Resultsmentioning

confidence: 99%

Provisional Use of CLSI-Approved Quality Control Strains for Antimicrobial Susceptibility Testing of Mycoplasma (‘Mesomycoplasma’) hyorhinis

et al. 2021

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“…To determine the analytical sensitivities of the multiplex qPCR, the three reference strains G. parasuis serovar 1, M. hyorhinis ATCC 17981 T , and M. hyosynoviae NCTC 10167 T were examined. With an estimated average genome size of 1.8 Mbp for G. parasuis (Brockmeier et al, 2014 ), 0.843 Mbp for M. hyorhinis (Cibulski et al, 2016 ; Käbisch et al, 2021 ; Trueeb et al, 2019 ), and 0.864 Mbp for M. hyosynoviae (GenBank accession number: CP008748.1), the following approximate DNA quantities corresponded to 1 genome equivalent (GE): 2 fg for G. parasuis , 0.95 fg for M. hyorhinis , and 0.93 fg for M. hyosynoviae . To obtain an accurate limit of detection (LoD) for each target and to identify a reasonable cut‐off C t value, 20 replicates of each reference strain were analyzed with concentrations close to the LoD ( G. parasuis : 1 GE, 10 GE, 100 GE, 200 GE, 500 GE, 2000 GE; vtaA : 1 GE, 100 GE, 200 GE, 1000 GE, 2000 GE, 5000 GE; M. hyorhinis : 1 GE, 20 GE, 100 GE, 200 GE, 300 GE, 1000 GE; M. hyosynoviae : 1 GE, 5 GE, 10 GE, 20 GE, 50 GE, 100 GE).…”

Section: Methodsmentioning

confidence: 99%

Section: Analytical Sensitivitymentioning

confidence: 99%

Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae

et al. 2023

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Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae are important porcine pathogens responsible for polyserositis, polyarthritis, meningitis, pneumonia, and septicemia causing significant economic losses in the swine industry. A new multiplex quantitative polymerase chain reaction (qPCR) was designed on one hand for the detection of G. parasuis and the virulence marker vtaA to distinguish between highly virulent and non-virulent strains. On the other hand, fluorescent probes were established for the detection and identification of both M. hyorhinis and M. hyosynoviae targeting 16S ribosomal RNA genes. The development of the qPCR was based on reference strains of 15 known serovars of G. parasuis, as well as on the type strains M. hyorhinis ATCC 17981 T and M.hyosynoviae NCTC 10167 T . The new qPCR was further evaluated using 21 G.parasuis, 26 M. hyorhinis, and 3 M. hyosynoviae field isolates. Moreover, a pilot study including different clinical specimens of 42 diseased pigs was performed. The specificity of the assay was 100% without cross-reactivity or detection of other bacterial swine pathogens. The sensitivity of the new qPCR was demonstrated to be between 11-180 genome equivalents (GE) of DNA for M. hyosynoviae and M. hyorhinis, and 140-1200 GE for G. parasuis and vtaA. The cut-off threshold cycle was found to be at 35. The developed sensitive and specific qPCR assay has the potential to become a useful molecular tool, which could be implemented in veterinary diagnostic laboratories for the detection and identification of G. parasuis, its virulence marker vtaA, M. hyorhinis, and M. hyosynoviae.

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“…The field isolates were chosen according to the year of isolation as well as the source of isolation (country, organs). Although erythromycin resistance is fairly common among porcine mycoplasmas, we evaluated the susceptibility of M. hyorhinis towards erythromycin to monitor the occurrence of elevated MICs in field isolates and not to overlook any possible changes [12][13][14]36].…”

Section: Antimicrobial Susceptibility Testing Of M Hyorhinis Field Is...mentioning

confidence: 99%

“…It is known that certain porcine mycoplasmas developed resistance against 14-membered macrolides, more specifically erythromycin, though the phenotypic manifestation is variable within field isolates [12]. The resistance is conferred by a point mutation within the 23S rRNA [13,14]. Correspondingly, the literature reports increased minimal inhibitory concentrations (MICs) for erythromycin among M. hyorhinis field isolates [6,8,9,15,16].…”

Section: Introductionmentioning

confidence: 99%

Towards a Standardized Antimicrobial Susceptibility Testing Method for Mycoplasma hyorhinis

et al. 2023

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Conducting antimicrobial susceptibility testing (AST) in a comparable manner requires the availability of a standardized method. Organizations, such as the Clinical and Laboratory Standards Institute (CLSI) or the European Committee on Antimicrobial Susceptibility Testing (EUCAST), provide standardized protocols for a range of fastidious bacteria but not for Mycoplasma hyorhinis. We developed a broth microdilution method for testing M. hyorhinis in a standardized and harmonized way using a modified Friis broth devoid of antimicrobial or otherwise bacterial growth-inhibiting agents. The type strain M. hyorhinis DSM 25591 was chosen to establish the methodology. The antimicrobial agents of interest were doxycycline, enrofloxacin, erythromycin, florfenicol, gentamicin, marbofloxacin, tetracycline, tiamulin, tilmicosin, tulathromycin, and tylosin, tested by using commercial SensititreTM microtiter plates. In addition, the suitability of the methodology was evaluated via variation of the individual ingredients of the modified Friis broth by either using different batches or choosing other distributors. Despite these alterations, the method provided reliable results. We obtained repeatable minimal inhibitory concentrations for all six tested field isolates and the M. hyorhinis type strain. With this newly proposed method, we aim to provide an improved AST method for diagnostic laboratories and monitoring purposes with better comparability between times and countries. In addition, this new method will allow for an improvement of targeted treatments using antimicrobial agents and thereby reduce the options for resistance development.

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Whole-Genome Sequence of the Mycoplasma ( Mesomycoplasma ) hyorhinis DSM 25591 Type Strain

Cited by 4 publications

References 15 publications

Provisional Use of CLSI-Approved Quality Control Strains for Antimicrobial Susceptibility Testing of Mycoplasma (‘Mesomycoplasma’) hyorhinis

Provisional Use of CLSI-Approved Quality Control Strains for Antimicrobial Susceptibility Testing of Mycoplasma (‘Mesomycoplasma’) hyorhinis

Development of a new multiplex quantitative PCR for the detection of Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae

Towards a Standardized Antimicrobial Susceptibility Testing Method for Mycoplasma hyorhinis

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