VETERINARY MICROBIOLOGY
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2021
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IF 3.293
Jingjing Qi,Yu Wang,Haoran Li,Yuanbing Shang,Song Gao,Chan Ding,Xiaohan Liu,Shaohui Wang,Tao Li,Mingxing Tian,Shengqing Yu
Mycoplasma synoviae ( M. synoviae ) is an important avian pathogen that causes arthritis and airsacculitis in young chickens and turkeys. Infection by M. synoviae results in considerable economic losses to the poultry industry worldwide. Cytoadherence is a crucial stage during mycoplasma infection. Dihydrolipoamide dehydrogenase (PdhD) is a flavin-dependent enzyme that is critical for energy metabolism and redox balance. To date, its role in cytoadherence is poorly understood. In this study, recombinant PdhD from M. synoviae (rMSPdhD) was expressed in the supernatant component of E. coli BL21 and rabbit anti-rMSPdhD serum was prepared. rMSPdhD was shown to be an immunogenic protein by immunoblot assays, while the mycoplasmacidal assay revealed that the rabbit anti-rMSPdhD serum had a high complement-dependent mycoplasmacidal rate (88.5 %). Using a suspension immunofluorescence assay and subcellular localization analysis, MSPdhD was shown to be a surface-localized protein distributed in both the cytoplasm and cell membrane of M. synoviae . The enzymatic activity of rMSPdhD was determined by measuring its ability to reduce lipoamide to dihydrolipoamide and convert NADH to NAD + . Using an indirect immunofluorescence assay, rMSPdhD was shown to adhere to DF-1 chicken embryo fibroblast cells. Furthermore, the attachment of M. synoviae to DF-1 cells was significantly inhibited by rabbit anti-rMSPdhD serum. Western blot and ELISA binding assays confirmed that rMSPdhD also bound to fibronectin (Fn) and plasminogen (Plg) in a dose-dependent manner. In conclusion, our data show that MSPdhD is not only a biological enzyme, but also an immunogenic surface-exposed protein that can bind to Fn and Plg as well as adhere to host cells. In addition, we show that rabbit anti-rMSPdhD serum can inhibit the adhesion of M. synoviae to DF-1 cells and has a significant complement-dependent bactericidal activity. Our findings suggest that MSPdhD may be involved in the pathogenesis of M. synoviae .
研究领域:
微生物学
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免疫学
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分子生物学
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DOI
PubMed
高影响因子
Transboundary and Emerging Diseases
·
2026
·Vol. 2026
·期 1
·2299833
·
IF 2.6
Qinghua Wang,Dan Li,Yue Wu,Yang Zhang,Dongjing Wang,Xiuwu Lian,Zhongshun Huang,Huiying Li,Yunchong Ma,Yilin Li,Jiangyong Zeng,Lijing Cao,Honglei Ding
Mycoplasma synoviae infection causes infectious synovitis and respiratory disease in broilers and a reduction in egg production and eggshell apex abnormalities (EAA) in layers, with widespread prevalence in the Chinese mainland since 2010. However, data on this pathogen in Chongqing remain scarce. The present study aimed to investigate the prevalence, sequence types (STs), hemagglutination activity, pathogenicity, and susceptibility to antimicrobials and Chinese herbal monomers of M. synoviae isolates from broilers and layers mainly in Chongqing. Additionally, we analyzed the genotype–phenotype correlations underlying the resistance of these isolates to fluoroquinolones and macrolides. A total of 23 M. synoviae isolates were recovered from 148 Chinese indigenous broilers, and 22 isolates were obtained from 210 eggs with EAA, yielding 45 isolates totally. These isolates were classified into five novel STs, namely ST196, ST258, ST261, ST264, and ST265. Phylogenetic analysis indicated that isolates belonging to the same ST formed distinct monophyletic clusters, and all isolates shared the closest genetic affinity with strains previously isolated in China. Of the 45 isolates, 42.2% were able to agglutinate chicken erythrocytes. All isolates were susceptible to doxycycline, tylvalosin, tilmicosin, lincomycin, and tiamulin but uniformly resistant to enrofloxacin. The majority of isolates displayed minimum inhibitory concentration (MIC) values >4 μg/mL for amikacin (97.8%), apramycin (93.3%), and erythromycin (91.1%). Notably, the Chinese herbal monomer berberine hydrochloride exhibited potent anti- M. synoviae activity, with an MIC range of 2–32 μg/mL. Genotypic analysis identified mutations in the parC gene (A56T, T85I, N89D) and ribosomal protein L4 (I174V, N215S), which were associated with the resistance of isolates to enrofloxacin and erythromycin, respectively. Both joint- and egg-derived isolates induced infectious synovitis and caseous exudates in specific pathogen-free (SPF) chickens. These findings highlight M. synoviae genetic/phenotypic diversity in Chongqing, emphasizing the need for rational antimicrobial stewardship to curb drug-resistant M. synoviae isolates.
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