Supplementary data – M. iowae antimicrobial resistance genetic markers – Buni, 2026 (hdl:21.15109/ARP/QM0ZUO)

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Supplementary data – M. iowae antimicrobial resistance genetic markers – Buni, 2026

hdl:21.15109/ARP/QM0ZUO

ARP

2026-06-12

1

Buni, Dominika; Kovács, Áron Botond; Wehmann, Enikő; Grózner, Dénes; Bányai, Krisztián; Nagy, Eszter Zsófia; Bradbury, Janet; Bottinelli, Marco; Stefani, Elisabetta; Catania, Salvatore; Lysnyansky, Inna; Kovács, László; Gyuranecz, Miklós; Kreizinger, Zsuzsa, 2026, "Supplementary data – M. iowae antimicrobial resistance genetic markers – Buni, 2026", https://hdl.handle.net/21.15109/ARP/QM0ZUO, ARP, V1

Supplementary data – M. iowae antimicrobial resistance genetic markers – Buni, 2026

hdl:21.15109/ARP/QM0ZUO

Buni, Dominika (Veterinary Medical Research Institute)

Kovács, Áron Botond (Veterinary Medical Research Institute)

Wehmann, Enikő (Veterinary Medical Research Institute)

Grózner, Dénes (Veterinary Medical Research Institute)

Bányai, Krisztián (Veterinary Medical Research Institute)

Nagy, Eszter Zsófia (Veterinary Medical Research Institute)

Bradbury, Janet (Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool)

Bottinelli, Marco (Istituto Zooprofilattico Sperimentale delle Venezie)

Stefani, Elisabetta (Istituto Zooprofilattico Sperimentale delle Venezie)

Catania, Salvatore (Istituto Zooprofilattico Sperimentale delle Venezie)

Lysnyansky, Inna (Department of Avian Diseases, Kimron Veterinary Institute)

Kovács, László (University of Veterinary Medicine Budapest)

Gyuranecz, Miklós (Veterinary Medical Research Institute)

Kreizinger, Zsuzsa (Veterinary Medical Research Institute)

FK21 (137809)

TKP2021-EGA-01

SA-27/2021

RRF-2.3.1-21-2022-00001

RRF-2.3.1-21-2022-00006

ARP

Kreizinger, Zsuzsa

Kaján, Győző László

2026-06-11

https://hdl.handle.net/21.15109/ARP/QM0ZUO

Agricultural Sciences, Mycoplasma iowae, Antimicrobial resistance, Efflux pumps, Mismatch amplification mutation assay (MAMA), Orthovanadate, Turkey pathogen

<i>Mycoplasma iowae</i> is an economically significant pathogen that causes reduced hatchability, late embryo mortality and leg deformities, chondrodystrophy and skeletal lesions in poults. While prevention is essential in the control of infection, the appropriate administration of antibiotics may reduce economic losses during outbreaks. As a first step in the exploration of antimicrobial resistance mechanisms in <i>M. iowae</i>, target modification and efflux pump activity were examined in the present study. Point mutations were analyzed in previously described antibiotic binding sites in the whole genome sequences of 99 <i>M. iowae</i> strains. Mismatch amplification mutation assays (MAMAs) were designed and validated for the differentiation of mutations corresponding to elevated minimum inhibitory concentration (MIC) values for fluoroquinolones. Broth microdilution assays were performed to evaluate the effect of efflux pump inhibitors. In the presence of orthovanadate (OV), MIC values were significantly lower than in the absence of OV for spiramycin, tilmicosin, tylosin and oxytetracycline, which may indicate the presence of an active efflux system in <i>M. iowae</i>. Putative promoter regions of efflux-related genes were predicted and characterized. Genetic mutations, previously described in other bacteria, were described to be associated with elevated fluoroquinolone, macrolide and lincomycin MICs in <i>M. iowae</i>, although certain resistant phenotypes remained unexplained, promoting future examinations for deeper insights. The developed MAMAs may support rapid identification of <i>M. iowae</i> strains with elevated MIC values for fluoroquinolones. The better understanding of the efflux pump mechanisms enables the development of alternative methods for the support of therapy against this pathogen.

<a href="http://creativecommons.org/licenses/by-nc/4.0">CC BY-NC 4.0</a>

NCBI BioProject ID: PRJNA975348

Identification and detection of genetic markers associated with antimicrobial susceptibility and evaluation of efflux pump mechanisms in Mycoplasma iowae Buni D, Kovács ÁB, Wehmann E, Grózner D, Bányai K, et al. (2026) Identification and detection of genetic markers associated with antimicrobial susceptibility and evaluation of efflux pump mechanisms in Mycoplasma iowae. PLOS ONE 21(4): e0347345

10.1371/journal.pone.0347345

Identification and detection of genetic markers associated with antimicrobial susceptibility and evaluation of efflux pump mechanisms in Mycoplasma iowae Buni D, Kovács ÁB, Wehmann E, Grózner D, Bányai K, et al. (2026) Identification and detection of genetic markers associated with antimicrobial susceptibility and evaluation of efflux pump mechanisms in Mycoplasma iowae. PLOS ONE 21(4): e0347345

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readme.md

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readme file

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table_s1.xlsx

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This table contains information on 99 *M. iowae* clinical isolates and strains, including strain designation, country of origin, host species, serotype, minimum inhibitory concentrations (MIC) for enrofloxacin, erythromycin, spiramycin, tilmicosin, tylosin, and lincomycin. It also lists the non-synonymous single nucleotide polymorphisms (nsSNPs) identified in the *gyrA*, *parC* and 23S rRNA genes associated with antimicrobial resistance.

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table_s1_sheet_1.csv

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This table contains information on 99 *M. iowae* clinical isolates and strains, including strain designation, country of origin, host species, serotype, and minimum inhibitory concentrations (MIC) for enrofloxacin. It also lists the non-synonymous single nucleotide polymorphisms (nsSNPs) identified in the *gyrA* and *parC* genes associated with antimicrobial resistance.

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table_s1_sheet_2.csv

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This table contains information on 99 *M. iowae* clinical isolates and strains, including strain designation, country of origin, host species, serotype, minimum inhibitory concentrations (MIC) for erythromycin, spiramycin, tilmicosin, tylosin, and lincomycin. It also lists the non-synonymous single nucleotide polymorphisms (nsSNPs) identified in the 23S rRNA genes associated with antimicrobial resistance.

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table_s2.csv

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This table presents non-synonymous SNPs identified in putative efflux pump-related genes in *M. iowae* strains used for efflux pump investigation. It includes the MIC values of enrofloxacin, lincomycin, oxytetracycline, spiramycin, tilmicosin and tylosin measured both in the absence and presence of sodium orthovanadate (OV). The data supports the evaluation of efflux pump-mediated antimicrobial resistance mechanisms.

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table_s2.xlsx

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This table presents non-synonymous SNPs identified in putative efflux pump-related genes in *M. iowae* strains used for efflux pump investigation. It includes the MIC values of enrofloxacin, lincomycin, oxytetracycline, spiramycin, tilmicosin and tylosin measured both in the absence and presence of sodium orthovanadate (OV). The data supports the evaluation of efflux pump-mediated antimicrobial resistance mechanisms.

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table_s3.csv

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Data used for logistic regression and Fisher's exact test to analyze associations among MIC values and genotypes in *Mycoplasma iowae*.

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table_s3.xlsx

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Data used for logistic regression and Fisher's exact test to analyze associations among MIC values and genotypes in *Mycoplasma iowae*.

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table_s4.csv

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Data used for Wilcoxon Signed-Rank test to analyze the effect of sodium orthovanadate on MIC values against *Mycoplasma iowae* strains.

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table_s4.xlsx

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Data used for Wilcoxon Signed-Rank test to analyze the effect of sodium orthovanadate on MIC values against *Mycoplasma iowae* strains.

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table_s5.xlsx

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Results of the logistic regression analysis and Fisher's exact test regarding associations among MIC values and genotypes in *Mycoplasma iowae*.

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table_s5_sheet_1.csv

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Contains chi-square statistics, p-values, odds ratios, coefficient estimates, and McFadden R² for the association between MIC values and genotypes at *gyrA*, *parC*, and 23S rRNA loci.

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table_s5_sheet_2.csv

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Provides contingency table analyses with corresponding odds ratios, p-values, and 95% confidence intervals for each antibiotic-genotype combination.

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table_s6.csv

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Results of the Wilcoxon Signed-Rank test regarding the effect of sodium orthovanadate on MIC values against *Mycoplasma iowae* strains.

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table_s6.xlsx

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Results of the Wilcoxon Signed-Rank test regarding the effect of sodium orthovanadate on MIC values against *Mycoplasma iowae* strains.

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