• Korean Journal of Veterinary Research
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• v.35 no.2
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• pp.317-326
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• 1995

The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

• Korean Journal of Veterinary Research
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• v.31 no.1
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• pp.71-75
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• 1991

This study was undertaken to identify the distribution of T and B lymphocytes in bovine peripheral blood and various lymphoid tissues by the method of ABPC using RABTS, $BLT_1$ and $_6E_{12}$ as primary antibodies. RABTS, $BLT_1$ and $_6E_{12}$ positive cells in PB-MNCs were 70.9${\pm}5.5%$, $59.0{\pm}8.7%$ and $23.0{\pm}8.7%$, respectively. $BLT_1$ and $_6E_{12}$ positive cells in nylon wool nonadherent cells of PB-MNCs were $91.6{\pm}1.0%$ and $9.6{\pm}0.8%$, respectively. In the lymphoid tissues such as inguinal lymph node, mesenteric lymph node, spleen and thymus, positive cells of RABTS were $76.3{\pm}3.4%$, $74.2{\pm}8.2%$, $73.6{\pm}5.5%$ and $95.6{\pm}2.8%$, those of $BLT_1$ were $56.4{\pm}6.2%$, $55.6{\pm}7.7%$, $48.6{\pm}5.1%$ and $23.0{\pm}4.8%$ and those of $_6E_{12}$ were $45.3{\pm}7.4%$, $42.3{\pm}5.8%$, $48.5{\pm}6.2%$ and $5.6{\pm}2.1%$, in order. These results are indicating that nylon wool column method is effective for separation of bovine ocytes.