Supplementary data – M. iowae antimicrobial resistance genetic markers – Buni, 2026hdl:21.15109/ARP/QM0ZUOARP2026-06-121Buni, 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, V1Supplementary data – M. iowae antimicrobial resistance genetic markers – Buni, 2026hdl:21.15109/ARP/QM0ZUOBuni, DominikaKovács, Áron BotondWehmann, EnikőGrózner, DénesBányai, KrisztiánNagy, Eszter ZsófiaBradbury, JanetBottinelli, MarcoStefani, ElisabettaCatania, SalvatoreLysnyansky, InnaKovács, LászlóGyuranecz, MiklósKreizinger, ZsuzsaFK21 (137809)TKP2021-EGA-01SA-27/2021RRF-2.3.1-21-2022-00001RRF-2.3.1-21-2022-00006ARPKreizinger, ZsuzsaKaján, Győző László2026-06-11Agricultural SciencesMycoplasma iowaeAntimicrobial resistanceEfflux pumpsMismatch amplification mutation assay (MAMA)OrthovanadateTurkey 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: PRJNA975348Identification 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): e034734510.1371/journal.pone.0347345Identification 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): e0347345readme.mdreadme filetext/markdowntable_s1.xlsxThis 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.application/vnd.openxmlformats-officedocument.spreadsheetml.sheettable_s1_sheet_1.csvThis 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.text/csvtable_s1_sheet_2.csvThis 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.text/csvtable_s2.csvThis 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.text/csvtable_s2.xlsxThis 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.application/vnd.openxmlformats-officedocument.spreadsheetml.sheettable_s3.csvData used for logistic regression and Fisher's exact test to analyze associations among MIC values and genotypes in *Mycoplasma iowae*.text/csvtable_s3.xlsxData used for logistic regression and Fisher's exact test to analyze associations among MIC values and genotypes in *Mycoplasma iowae*.application/vnd.openxmlformats-officedocument.spreadsheetml.sheettable_s4.csvData used for Wilcoxon Signed-Rank test to analyze the effect of sodium orthovanadate on MIC values against *Mycoplasma iowae* strains.text/csvtable_s4.xlsxData used for Wilcoxon Signed-Rank test to analyze the effect of sodium orthovanadate on MIC values against *Mycoplasma iowae* strains.application/vnd.openxmlformats-officedocument.spreadsheetml.sheettable_s5.xlsxResults of the logistic regression analysis and Fisher's exact test regarding associations among MIC values and genotypes in *Mycoplasma iowae*.application/vnd.openxmlformats-officedocument.spreadsheetml.sheettable_s5_sheet_1.csvContains 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.text/csvtable_s5_sheet_2.csvProvides contingency table analyses with corresponding odds ratios, p-values, and 95% confidence intervals for each antibiotic-genotype combination.text/csvtable_s6.csvResults of the Wilcoxon Signed-Rank test regarding the effect of sodium orthovanadate on MIC values against *Mycoplasma iowae* strains.text/csvtable_s6.xlsxResults of the Wilcoxon Signed-Rank test regarding the effect of sodium orthovanadate on MIC values against *Mycoplasma iowae* strains.application/vnd.openxmlformats-officedocument.spreadsheetml.sheet