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实验性接种猪中猪肺炎支原体232菌株感染的动态变化和时间顺序

Dynamics and chronology of Mycoplasma hyopneumoniae strain 232 infection in experimentally inoculated swine.

作者信息

Almeida Henrique M S, Mechler-Dreibi Marina L, Sonálio Karina, Ferreira Marcela M, Martinelli Paulo E B, Gatto Igor R H, Maes Dominiek, Montassier Hélio J, Oliveira Luís G

机构信息

São Paulo State University (Unesp), School of Agricultural and Veterinarian Sciences, Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP, 14884-900, Brazil.

Faculty of Veterinary Medicine, Ghent University, Ghent, Belgium.

出版信息

Porcine Health Manag. 2021 Jun 30;7(1):42. doi: 10.1186/s40813-021-00221-2.

Abstract

Direct detection of Mycoplasma hyopneumoniae through molecular tools is a growing trend for early diagnosis, highlighting the importance of knowing M. hyopneumoniae dynamics in the respiratory tract upon infection. This study focused on monitoring the infection level and its effects in different anatomic sites of the respiratory tract of experimentally infected swine in four time-points post-infection. To this end, 24 pigs were allocated to either non-inoculated group (n = 8) or inoculated group (n = 16). On day 0 post-infection (dpi), animals of the inoculated group were intratracheally inoculated with M. hyopneumoniae. Nasal swabs were collected weekly for qPCR detection of bacterial shedding. At 14, 28, 42, and 56 dpi, four animals from the inoculated group and two from the control group were necropsied. Bronchoalveolar lavage fluid (BALF) and samples from three different anatomical tracheal sections (cranial - CT, medium - MT, lower - LT) were collected for qPCR and histopathology. Bacterial loads (qPCR) in tracheal samples were: 4.47 × 10 copies∕μL (CT), 1.5 × 10- copies∕ μL (MT) and 1.4 × 10 copies∕μL (LT samples). M. hyopneumoniae quantification in BALF showed the highest load at 28 dpi (2.0 × 10 copies∕ μL). Microscopic lesions in LT samples presented the highest scores at 56 dpi and were significantly correlated with the pathogen load on 14 dpi (0.93) and 28 dpi (0.75). The greatest bacterial load of M. hyopneumoniae in CT samples and BALF was registered at 28 dpi, and it remained high in BALF and LT throughout the 56 dpi. The pathogen was able to persist during the whole experimental period, however higher estimated quantification values were registered in the lower parts of the respiratory tract, especially at 56 dpi. These findings are important for improving diagnostics, treatment, and control measures of M. hyopneumoniae infection in swine herds.

摘要

通过分子工具直接检测猪肺炎支原体是早期诊断的一个发展趋势,这凸显了了解感染后呼吸道中猪肺炎支原体动态变化的重要性。本研究着重于监测感染后四个时间点实验感染猪呼吸道不同解剖部位的感染水平及其影响。为此,将24头猪分为未接种组(n = 8)和接种组(n = 16)。在感染后第0天(dpi),给接种组动物进行气管内接种猪肺炎支原体。每周采集鼻拭子用于qPCR检测细菌排出情况。在感染后14、28、42和56天,对接种组的4头动物和对照组的2头动物进行剖检。收集支气管肺泡灌洗液(BALF)以及来自三个不同解剖气管段(头段 - CT、中段 - MT、下段 - LT)的样本用于qPCR和组织病理学检查。气管样本中的细菌载量(qPCR)分别为:4.47×10拷贝∕μL(CT段)、1.5×10⁻拷贝∕μL(MT段)和1.4×10拷贝∕μL(LT段样本)。BALF中猪肺炎支原体的定量分析显示在感染后28天载量最高(2.0×10拷贝∕μL)。LT段样本的微观病变在感染后56天得分最高,并且与感染后14天(0.93)和28天(0.75)的病原体载量显著相关。CT段样本和BALF中猪肺炎支原体的最大细菌载量出现在感染后28天,并且在整个56天的观察期内,BALF和LT段中的载量一直保持较高水平。病原体能够在整个实验期间持续存在,然而估计定量值在呼吸道下部更高,尤其是在感染后56天。这些发现对于改进猪群中猪肺炎支原体感染的诊断、治疗和控制措施具有重要意义。

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