Departamento de Ciencias Clínicas Veterinarias, Universidad de Santiago de Compostela, Lugo, Spain.
J Fish Dis. 2014 Nov;37(11):959-68. doi: 10.1111/jfd.12199. Epub 2013 Nov 26.
A sensitive and specific immunohistochemical technique was developed to improve the diagnosis of tenacibaculosis and to better understand its pathogenesis. Senegalese sole Solea senegalensis Kaup, 1858 were inoculated subcutaneously with a bacterial suspension of Tenacibaculum maritimum, and samples were taken at different hours post-inoculation. Sections from different organs were used as positive controls. In addition, a total of 128 field samples from different organs collected from tenacibaculosis outbreaks were used. Tenacibaculum maritimum antigens were detected in several organs of experimentally infected Senegalese sole and in at least one of the tissues from fish suffering from natural tenacibaculosis previously confirmed by culture and PCR-based methods. In fish collected during outbreaks, a strong positive reaction was detected in ulcerative skin areas. Moreover, bacterial antigen was identified inside scale pockets and in sites of the skin with mild lesion. In kidney and spleen, evident immunostaining of bacterial antigen was detected in both naturally and experimentally infected fish. Besides, the presence of T. maritimum in the intestinal tract without associated histological changes suggests that this organ may act as a reservoir for T. maritimum. The results of this study confirm the usefulness of IHC for the diagnosis of tenacibaculosis in paraffin-embedded tissues.
本研究建立了一种敏感且特异的免疫组化技术,以提高对迟钝爱德华氏菌病的诊断能力,并深入了解其发病机制。将迟钝爱德华氏菌悬浮液皮下接种塞内加尔鳎(Solea senegalensis Kaup, 1858),并在接种后不同时间点取样。使用来自不同器官的切片作为阳性对照。此外,共使用了 128 份来自不同器官的现场样本,这些样本来自已通过培养和基于 PCR 的方法证实的迟钝爱德华氏菌病暴发。在实验感染的塞内加尔鳎的几个器官中以及在先前通过培养和基于 PCR 的方法证实患有自然迟钝爱德华氏菌病的鱼类的至少一种组织中均检测到了迟钝爱德华氏菌抗原。在暴发期间采集的鱼类中,在溃疡性皮肤区域检测到强烈的阳性反应。此外,在鳞片袋和皮肤轻度病变部位也鉴定出了细菌抗原。在肾脏和脾脏中,无论是自然感染还是实验感染的鱼类,均检测到明显的细菌抗原免疫染色。此外,肠道中存在无相关组织学变化的 T. maritimum 表明该器官可能是 T. maritimum 的储存库。本研究的结果证实了免疫组化技术在石蜡包埋组织中诊断迟钝爱德华氏菌病的有用性。