Gayo E, Polledo L, Magalde A, Balseiro A, García Iglesias M J, Pérez Martínez C, Preziuso S, Rossi G, García Marín J F
Veterinary Pathology Unit, School of Veterinary Medicine, University of León, León, Spain.
Micros Veterinaria, León, Spain.
BMC Vet Res. 2019 Apr 10;15(1):109. doi: 10.1186/s12917-019-1855-3.
In order to characterize the complete range of lesions, especially minimal, affecting mammary gland and viral antigen distribution and target cells using immunohistochemistry in naturally Visna/maedi (VM) 84 infected sheep were studied, forty-four from flocks with clinical cases (A) and 35 randomly sampled from two abattoirs (B) together with five negative controls (C). An immunocytochemistry technique was developed and further milk samples (n = 39) were used to study viral excretion, carrier cells and the role of milk and colostrum in the transmission of the disease.
All sheep from group C and three sheep from group B were negative to VM in tissue sections by histopathology, immunohistochemistry and PCR, and also in serum using ELISA. Several degrees of CD3 + lymphocytic interstitial mastitis were observed in groups A and B: minimal (+) n = 26 sheep; moderate (++), n = 32 and severe (+++), n = 12. No differences in lesion distribution were observed between groups A and B. Viral presence was confirmed by immunohistochemistry using two different antibodies and/or PCR in every tissue with lesions while serology was negative in six sheep with lesions. Two milk samples taken from milk tanks from two flocks from group A and fourteen milk samples from 29 infected sheep from group B were positive to VM (most of them from animals with moderate and severe lesions). Positivity was only found in macrophages, even in focal and minimal lesions, while no positivity was observed in epithelial or any other cells in either tissue and milk samples.
This new observation of the minimal lesions described in this work increased the prevalence of VM lesions in mammary gland up to 90.9% and VM should be considered as a differential diagnosis when minimal interstitial lesions are detected. A high prevalence of VM was observed in intensive milk-producing sheep, ELISA serology did not detect as positivity all infected animals, while histology, IHC or PCR showed higher sensitivity. The cytological technique developed was very useful in milk-cell studies using hematoxylin and eosin and immunocytochemistry. Viral detection in milk samples (16/39) confirms a potential but limited role of milk/colostrum in viral transmission.
为了通过免疫组织化学来描述自然感染维斯纳/梅迪病(VM)的绵羊乳腺病变的完整范围,尤其是微小病变,以及病毒抗原分布和靶细胞,对44只来自有临床病例的羊群(A组)、35只从两个屠宰场随机抽取的绵羊(B组)以及5只阴性对照绵羊(C组)进行了研究。开发了一种免疫细胞化学技术,并进一步使用牛奶样本(n = 39)来研究病毒排泄、携带细胞以及牛奶和初乳在疾病传播中的作用。
通过组织病理学、免疫组织化学和PCR检测,C组所有绵羊以及B组3只绵羊的组织切片对VM呈阴性,血清ELISA检测也呈阴性。在A组和B组中观察到了不同程度的CD3 +淋巴细胞性间质性乳腺炎:轻微(+),n = 26只绵羊;中度(++),n = 32只;重度(+++),n = 12只。A组和B组之间在病变分布上未观察到差异。使用两种不同抗体的免疫组织化学和/或PCR在每例有病变的组织中均证实了病毒的存在,而6只患有病变的绵羊血清学检测呈阴性。从A组两个羊群的奶罐中采集的两份牛奶样本以及从B组29只感染绵羊中采集的14份牛奶样本对VM呈阳性(其中大多数来自患有中度和重度病变的动物)。仅在巨噬细胞中发现阳性,即使在局灶性和微小病变中也是如此,而在组织和牛奶样本的上皮细胞或任何其他细胞中均未观察到阳性。
本研究中对微小病变的新观察将VM在乳腺中的病变患病率提高到了90.9%,当检测到微小间质性病变时,VM应被视为鉴别诊断。在集约化产奶绵羊中观察到VM的高患病率,ELISA血清学检测并未检测出所有感染动物呈阳性,而组织学、免疫组织化学或PCR显示出更高的敏感性。所开发的细胞学技术在使用苏木精和伊红以及免疫细胞化学的乳汁细胞研究中非常有用。牛奶样本中的病毒检测(16/39)证实了牛奶/初乳在病毒传播中具有潜在但有限的作用。