Supplementary data – Mycoplasma hyorhinis vaccine strain – Nagy, 2026

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Nagy, Eszter Zsófia; Szeredi, Levente; Földi, Dorottya; Belecz, Nikolett; Kovács, Áron Botond; Sulyok, Kinga Mária; Grózner, Dénes; Wehmann, Enikő; Bányai, Krisztián; Marton, Szilvia; Tenk, Miklós; Kreizinger, Zsuzsa; Gyuranecz, Miklós, 2026, "Supplementary data – Mycoplasma hyorhinis vaccine strain – Nagy, 2026", https://hdl.handle.net/21.15109/ARP/3IIFGY, ARP, V1
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Description
Background Mycoplasma (M.) hyorhinis causes substantial economic losses in swine. Currently, prevention and treatment rely on minimizing risk factors and administering antibiotics, as no vaccines are commercially available in Europe. However, antibiotics often cannot fully eliminate the bacteria. The development of an effective vaccine could lead to a potentially long-term control method. Materials and methods A temperature-sensitive M. hyorhinis strain was developed using 1-methyl-3-nitro-1-nitrosoguanidine (NTG) mutagenesis. The immunogenicity and efficacy of this vaccine candidate clone were evaluated in combination with an adjuvant. Three-week-old piglets were immunized with the candidate vaccine strain, and the vaccination site was monitored daily. At six weeks of age, the pigs were challenged intravenously on two subsequent days. Daily clinical examinations were conducted, with blood and nasal swabs collected weekly throughout the study for M. hyorhinis enzyme-linked immunosorbent assay (ELISA), real-time PCR analysis, and isolation. Three weeks post-challenge, the animals were euthanized for gross and histopathological examinations. Body temperature was recorded daily, and body weight was measured upon arrival, and then at six and nine weeks of age. Results Vaccination significantly reduced clinical signs (p = 0.03), as well as gross pathological (p = 0.01) and histopathological (p = 0.005) lesions compared with the positive control group. The vaccinated group exhibited an earlier and higher increase in M. hyorhinis-specific IgG antibody levels post-challenge than the positive control group. However, the vaccine candidate did not mitigate the impact of M. hyorhinis infection on the weight gain. After the challenge (days 21–42), both the vaccinated (p = 0.001) and the positive control (p = 0.003) groups exhibited reduced weight gain compared with the negative control group. Discussion Overall, the attenuated M. hyorhinis strain, combined with the adjuvant, provided protection against M. hyorhinis infection. These results form a basis for the development of a novel vaccine candidate that offers effective prevention. (2026-03-25)
Subject Agricultural Sciences
Keyword 1-Methyl-3-nitro-1-nitrosoguanidine, Live attenuated vaccine, Mycoplasma hyorhinis, NTG, Porcine, Temperature-sensitive strain
Related Publication Is Supplement To: Nagy EZ, Szeredi L, Földi D, et al. Development and efficacy test of a live, attenuated Mycoplasma hyorhinis vaccine candidate strain. Vaccine. 2026;75:128278. doi:10.1016/j.vaccine.2026.128278doi: 10.1016/j.vaccine.2026.128278
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supplementary_tables_1-14.xlsx
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Published Mar 25, 2026
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Supplementary tables 1-14
supplementary_table_10.csv
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Supplementary table 10. Gross pathological lesion scores of the pigs in the vaccinated, positive control, and negative control groups. Lesions were not observed in the negative control group.
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supplementary_table_11.csv
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Supplemetnary table 11. Histopathological lesion scores of the pigs in the vaccinated, positive control, and negative control groups.
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supplementary_table_12.csv
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Supplementary table 12. Optical density (OD) values of the Mycoplasma hyorhinis in-house ELISA in the vaccinated, positive control, and negative control groups.
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supplementary_table_13.csv
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Supplementary table 13. Results of the Mycoplasma hyorhinis qPCR from the samples taken during the gross pathological examination from the vaccinated, positive control, and negative control groups. The table also presents the quantification cycle (Cq) values of the positive samples and the organs where pathological lesions were observed (highlighted with a *).
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supplementary_table_14.csv
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Supplementary table 14. Results of Mycoplasma hyorhinis isolation and multiple-locus variable-number tandem-repeat analysis (MLVA). MLVA was performed following established protocols.
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supplementary_table_1A.csv
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Supplementary table 1A. Mastermix compositions of the applied qPCR assays.
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supplementary_table_1B.csv
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Supplementary table 1B. Thermal protocols of the applied qPCR assays.
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supplementary_table_2.csv
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Supplementary table 2. Scoring system for the clinical signs.
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supplementary_table_3.csv
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Supplementary table 3. Scoring system for the pathological lesions and evaluation of the vaccination site.
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