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福建地区滑液囊支原体的分离、鉴定及对7日龄SPF鸡致病性

王晨燕 侯博

王晨燕,侯博. 福建地区滑液囊支原体的分离、鉴定及对7日龄SPF鸡致病性 [J]. 福建农业学报,2023,38(4):436−443 doi: 10.19303/j.issn.1008-0384.2023.04.007
引用本文: 王晨燕,侯博. 福建地区滑液囊支原体的分离、鉴定及对7日龄SPF鸡致病性 [J]. 福建农业学报,2023,38(4):436−443 doi: 10.19303/j.issn.1008-0384.2023.04.007
WANG C Y, HOU B. Isolation, identification and pathogenicity of Mycoplasma synoviae strains from Fujian to 7-day-old SPF chickens [J]. Fujian Journal of Agricultural Sciences,2023,38(4):436−443 doi: 10.19303/j.issn.1008-0384.2023.04.007
Citation: WANG C Y, HOU B. Isolation, identification and pathogenicity of Mycoplasma synoviae strains from Fujian to 7-day-old SPF chickens [J]. Fujian Journal of Agricultural Sciences,2023,38(4):436−443 doi: 10.19303/j.issn.1008-0384.2023.04.007

福建地区滑液囊支原体的分离、鉴定及对7日龄SPF鸡致病性

doi: 10.19303/j.issn.1008-0384.2023.04.007
基金项目: 福建省属科技计划公益类专项(2022R10260014);福建省农业高质量发展超越“5511”协同创新工程项目(XTCXGC2021008);福建省农业科学院科技创新团队建设项目(CXTD2021014-3)
详细信息
    作者简介:

    王晨燕(1988−),女,硕士,助理研究员,主要从事家禽传染病病原学与防治技术研究(E-mail:15396022617@163.com

    通讯作者:

    侯博(1986−),男,博士,副研究员,主要从事家禽传染病病原学与防治技术研究(E-mail:houbohouwei@126.com

  • 中图分类号: S852.62

Isolation, identification and pathogenicity of Mycoplasma synoviae strains from Fujian to 7-day-old SPF chickens

  • 摘要:   目的  了解福建地区滑液囊支原体(Mycoplasma synoviae,MS)临床感染菌株的进化关系以及致病能力。  方法  对福建地区疑似MS感染鸡群的上颚裂和气管拭子样品进行病原检测、分离和鉴定,获得MS临床分离株,并进行vlhA基因进化树分析;选取其中6株MS分离株,通过点眼滴鼻途径感染7日龄SPF鸡,观察感染后的临床症状和解剖病理变化、气管组织病理损伤、气管病原再分离以及MS抗体阳性率,比较不同菌株的致病力和水平传播能力。  结果  气管拭子的MS检出率显著高于上颚裂拭子。通过分离、鉴定共获得9株MS分离株,遗传进化关系分析表明vlhA基因存在着多样性,不同分离株具有不同的进化来源。根据vlhA基因的进化关系,选择6株MS分离株感染7日龄SPF鸡,未观察到明显的临床症状;临床解剖发现,感染HI株14 d、感染 HI和SD6株21 d各有1羽鸡出现了气囊炎,而其余各鸡以及同居对照鸡均未发现有明显病理变化。气管组织病理学分析发现,不同MS分离株在感染后7、14、21 d的病理损伤能力存在显著差异;气管MS病原再分离结果显示,不同分离株在气管的定殖和复制能力存在明显差异,并且SD19和SD6株还具有较强的水平传播能力。MS抗体检测发现仅在感染后21 d SD19株感染组出现1羽感染鸡和1羽同居对照鸡抗体转阳。  结论  气管样品更适合MS的检测和分离,福建MS分离株不易引起7日龄SPF鸡临床解剖病理变化,但可在气管中持续存在和定殖,造成病理损伤,且不同菌株之间存在显著差异。本研究通过对不同MS菌株在7日龄SPF鸡中的致病力和水平传播能力的比较为后续开展滑液囊支原体病的防控技术研究奠定了基础。
  • 图  1  MS分离株的分离与鉴定

    A——KM2液体培养基中培养18~20 h的原代培养物的PCR检测结果,M:DNA标记物,1 ~ 10:临床样品扩增结果,11:PCR阴性对照,12:PCR阳性对照。B——MS分离菌株在KM2液体培养基中的生长情况,1:玫瑰红颜色的培养基(阴性对照),2 ~ 5:培养3 d后的培养基呈现橘黄色。C——滑液囊支原体HI株在KM2琼脂上呈“肚脐状”或“煎蛋状”菌落形态(50×)。

    Figure  1.  Isolation and identification of MS isolates

    A: Specificity of the PCR assay using specific primers performed on the primary culture in a KM2 medium for 18 to 20 h; M: DNA marker; 1 to10: cultures of clinical samples; 11: negative control; 12: positive control. B: Growth of MS isolates in KM2 medium; 1: Rose red color of the medium as negative control, 2 to 5: Orange color of medium cultivated MS strain after 3 days, C: Colonies of Mycoplasma synoviae HI strain on the KM2 agar (50×).

    图  2  MS 分离株与参考株的vlhA基因核苷酸序列遗传进化分析

    Figure  2.  Phylogenetic tree of nucleotide sequences between isolates and reference strains based on vlhA gene

    图  3  MS分离株HI感染SPF鸡14 d(B)和21 d(C)后气囊临床病理变化

    A:正常气囊薄而透明;B:气囊增厚,透明度下降(黑色箭头);C:气囊表面有大量气泡(黑色箭头)。

    Figure  3.  Clinical anatomical and pathological changes of SPF chickens air sac infected with HI strain for 14 (B) and 21 (C) days

    A: Normal air sac is thin and transparent; B: air sac is thickened (black arrow); C: air sac has foam on the surface (black arrow).

    图  4  MS HI株感染SPF鸡7、14和21 d后气管组织病理学损伤情况

    A、B和C分别为对照组7、14和21 d的正常气管组织结构。D、E和F分别为MS分离株HI感染后7、14和21 d的气管组织形态。

    Figure  4.  Histopathology lesions of trachea of 7-day-old SPF chickens infected with HI strain for 7, 14, and 21 days

    A–C: Normal tracheal structure of negative control chicken at 7, 14, and 21 days after infection; D–F: Morphological change of the trachea at 7, 14, and 21 days after infection of HI strain, respectively.

    图  5  不同MS分离株感染SPF鸡7 (A)、14 (B)和21 (C) d后气管组织病理学损伤评分(箱线图)

    NC为未感染的对照组,数据以非参数的Kruskal-Wallis统计方法进行检验,并进行多重比较,当P< 0.05时表示差异显著,以*表示。

    Figure  5.  Mean lesion scores of trachea of 7-day-old chickens infected with MS isolates for 7 (A), 14 (B) and 21 (C) days (Box-plot)

    NC: No challenge group(control). Data were analyzed using Kruskal-Wallis (for non-parametric data), followed by Dunn’s multiple comparison tests, * P< 0.05 indicates a significant difference.

    表  1  不同MS菌株对7日龄SPF鸡感染7、14、21 d后气管病原再分离结果

    Table  1.   Re-isolation of MS from trachea of 7-old day SPF chickens infected with different MS strains after 7, 14, and 21 days

    组别
    Group
    菌株
    Strain
    再分离率
    Results of re-isolation
    感染后7 d
    Re-isolation of
    7 dpi
    感染后14 d
    Re-isolation of
    14 dpi
    感染后21 d
    Re-isolation of
    21 dpi
    G1未接种No challenge0/60/7 0/7
    G2MS-SD180/6(0/3)*1/7(0/3)0/7(0/4)
    G3MS-HI3/6(1/3)5/7(0/3)5/7(1/4)
    G4MS-SD191/6(0/3)6/7(2/3)2/7(2/4)
    G5MS-SD170/6(0/3)3/7(0/3)5/7(1/4)
    G6MS-HB1/6(1/3)0/7(0/3)0/7(0/4)
    G7MS-SD64/6(0/3)2/7(3/3)5/7(1/4)
    *表中数据为阳性数/检测数(同居鸡的阳性数/检测数)。
    Data in the table was number of positive samples/number of tested samples, and number of positive samples/ number of tested samples for cohabitation chickens in parentheses.
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出版历程
  • 收稿日期:  2022-12-04
  • 修回日期:  2023-03-18
  • 网络出版日期:  2023-05-09
  • 刊出日期:  2023-04-28

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