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STUDIES ON TOXOPLASMOSIS V : COMPLEMENTAL OBSERVATIONS ON THE TISSUE CULTURE METHOD, ESPECIALLY ON THE EFFECT OF THE NUTRIENT FLUID UPON THE INVASION AND MULTIPLICATION OF THE ORGANISMS

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Please use this identifier to cite or link to this item:https://doi.org/10.14943/jjvr.11.1.1

Title: STUDIES ON TOXOPLASMOSIS V : COMPLEMENTAL OBSERVATIONS ON THE TISSUE CULTURE METHOD, ESPECIALLY ON THE EFFECT OF THE NUTRIENT FLUID UPON THE INVASION AND MULTIPLICATION OF THE ORGANISMS
Authors: SHIMIZU, Kiheiji Browse this author
Issue Date: Mar-1963
Publisher: The Graduate School of Veterinary Medicine
Journal Title: Japanese Journal of Veterinary Research
Volume: 11
Issue: 1
Start Page: 1
End Page: 11
Abstract: Following the previous report, complemental observations were carried out to ascertain the suitable cultural conditions including the effects of the nutrient fluid upon the invasion and multiplication, the ultimate aim being to promote the utility of tissue culture in isolation of the organisms. Throughout the experiments HeLa cells and T. gondii strain RH were used. Data obtained are summarized as follows : 1. Good invasion occured very frequently when Hanks' balanced salt solution and solutions devoid of divalent ions such as Mg, Ca, were employed. However, there seems to be no significant differences in the ratio of invasion due to the culture fluids tested-H (Hanks), HL (Hanks plus lactalbumin hydrolysate), HLS (HL plus serum), YLE (yeast extract, lactalbumin hydrolysate and Earle's balanced salt solution) and YLES (YLE plus serum), and organism numbers more than 90% may invade the host cell 24 hours after inoculation (fig. 1). 2. The organisms which were suspended in the culture fluid devoid of protein substances tend to die in rather short period and could not multiply well in cells when cultured by these proteinless nutrients (Fig. 3 A, Table 1). Accordingly, the nutrients must be replaced by ones containing protein after 5〜7 hours when the media such as Hanks were employed in inoculation (Fig. 3B). 3. Addition of bovine sera to the concentration of 0.5〜1.0% seems to be quite enough to cause heavy growth of T. gondii if the medium HL is used for the culture nutrient. 4. PH values ranging from 6.5 to 8.0 do seem not to show any effects on the invasion ratio of the organisms. However, in the more acid or alkaline sides (6.0,8.0) poor multiplication was always observed and especially in acid side, elution of the soluble complement-fixing antigenic substances was also scarce (Fig. 2). 5. The volume of the suspension of the inocula seemed to have no significant relations with the yield of the organisms when the inoculated bottles had been left overnight for the invasion (Fig. 7). 6. To cultivate a small number of the organisms in material, frequent changes of the nutrient fluid every day or every other day during the first week are very necessary (Fig. 6). 7. Clear proofs of the multiplication of the organism numbers of 2.5~5.0×10^3 in the materials were observed by counting method when the tests were performed with due consideration of the above described points (Fig. 8).
Type: bulletin (article)
URI: http://hdl.handle.net/2115/1772
Appears in Collections:Japanese Journal of Veterinary Research > Volume 11, Number 1

Submitter: 獣医学部図書室

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