Logan L L, Whyard T C, Quintero J C, Mebus C A
Department of Pathology, School of Veterinary Medicine, University of California, Davis 95616.
Onderstepoort J Vet Res. 1987 Sep;54(3):197-204.
The sequential development of C. ruminantium (Kwanyanga and Kümm isolates) was followed in caprine leukocyte cultures by light microscopy, direct immunofluorescent microscopy (DFA), indirect immunoflourescent microscopy (IFA) and transmission electron microscopy (TEM). During the febrile response, one to several small cocci, large ring forms or rods were observed in neutrophils in blood smears and cytopreparations of neutrophil fractions using Diff Quik stain, Giemsa stain, DFA and TEM. One to several C. ruminantium colonies were seen in up to 35% of neutrophils maintained in vitro for 18 h to 5 days. The organisms were located in neutrophil phagosomes by TEM and were enveloped by two trilamellar unit membranes. Initially, C. ruminantium was tightly enclosed within phagosomes. At 20 h of incubation, organisms were frequently observed undergoing binary fission within enlarged phagosomal vacuoles. At later time periods, neutrophils harboured fully formed colonies (morula) containing numerous organisms. An occasional C. ruminantium-infected macrophage (Kümm isolate), and an occasional infected eosinophil (Kümm and Kwanyanga isolate) were found.
通过光学显微镜、直接免疫荧光显微镜（DFA）、间接免疫荧光显微镜（IFA）和透射电子显微镜（TEM），在山羊白细胞培养物中追踪反刍兽无形体（夸扬加和库姆分离株）的连续发育过程。在发热反应期间，使用Diff Quik染色、吉姆萨染色、DFA和TEM，在血液涂片和中性粒细胞组分的细胞制备物中的中性粒细胞中观察到一至几个小球菌、大环形式或杆状。在体外培养18小时至5天的中性粒细胞中，高达35%的细胞中可见一至几个反刍兽无形体菌落。通过TEM观察到这些生物体位于中性粒细胞吞噬体中，并被两层三层单位膜包裹。最初，反刍兽无形体紧密包裹在吞噬体内。在孵育20小时时，经常观察到生物体在扩大的吞噬体空泡内进行二分裂。在后期，中性粒细胞中含有完全形成的菌落（桑葚胚），其中包含大量生物体。偶尔发现被反刍兽无形体感染的巨噬细胞（库姆分离株）和偶尔被感染的嗜酸性粒细胞（库姆和夸扬加分离株）。
