Studies on malarial sporozoites. II. Effect of age and dosage of sporozoites on their infectiousness.
Studies on malarial sporozoites. II. Effect of age and dosage of sporozoites on their infectiousness.
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疟疾子孢子的研究。
DOI：
10.1016/0014-4894(54)90026-0
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发表时间：
1954
影响因子：
2.1
通讯作者：
E. M. Dusseau
中科院分区：
文献类型：
--
作者：
R. J. Porter;R. Laird;E. M. Dusseau
材料和方法用子孢子感染小鸡的方法在别处已被广泛讨论（Porter，et al.，1952年）。Cedes aeg，i和A.白纹伊蚊是用Shannon和Putnam（1934）的一般方法饲养的。它们在羽化后一周内通过喂食携带8-A鸡疟原虫株血液通道的c，hiclts而感染。将受感染的蚊子保持在27-30 ℃（平均28 ℃）下，用煮熟的葡萄干或苹果片作为维持饮食。将头部切下，并将一点胸腔内容物挤出到0.85%的氯化钠溶液中。这样切除的组织通常包含唾液腺。将每批蚊子的腺体悬浮在0.50 ml肝素化的正常鸡血（0.85%盐水溶液，0.05 cc;粉末状肝素，约20 mg;血液，0.45 ml）中，在玻璃辊磨机中旋转30分钟。检查制备物的干燥、Giemsa染色的薄膜，以确定子孢子与红细胞的比率。至少计数10个子孢子，除非首先达到总共10，000个红细胞。这给出了一个数字（$子孢子/10，000红细胞），对于10或更大的值，可能误差为20%或更小。对于较小的值，概率误差逐渐增大。悬浮液中的子孢子总数可以从t计算，这是充分准确的假设，即0.45 ml正常鸡血含有约1，000，000，000个红细胞。每个悬浮液含有9至177个唾液腺，通常为10-20个蚊子。将制备物的等分试样静脉接种至2-7日龄的白色Rock或白色Leghorn雏鸡，每组4只或更多。
MATERIALS AND METHODSThe procedures for infection of chicks with sporozoites have been discussed extensively elsewhere (Porter, et al., 1952). Cedes aeg,@ i and, to a small extent, A. albopictus were reared by the general methods of Shannon and Putnam (1934). They were infected within a week of emergence by feeding on c, hiclts harboring a blood-passage line of the 8-A strain of Plasmodium gallinacewn. infected mosquitoes were kept at 27-30 C (mean 28”) with cooked raisins or slices of apple as a maintenance diet.For dissection, mosquitoes were immobilized with minimal exposure to anesthetic ether. The heads were cut off, and a bit of thoracic contents was squeezed out into 0.85% sodium chloride solution. The tissue thus removed usually contains the salivary glands. Glands from each lot of mosquitoes were suspended in 0.50 ml of heparinized normal chick blood (0.85% saline solution, 0.05 cc; powdered heparin, about 20 mg; blood, 0.45 ml) in a glass roller mill rotated for 30 minutes. Dried, Giemsa-stained films of the preparation were examined to determine the ratio of sporozoites to erythrocytes. At least 10 sporozoit, es were counted unless a total of 10,000 erythrocytes was reached first. This gives a figure ($ sporozoites per 10,000 erythrocytes) with a probable error of 20% or less for values of 10 or more. For smaller values, the probable error is progressively larger. The total number of sporozoites in the suspension ran be calculated from t, he sufficiently accurate assumption that 0.45 ml of normal chick blood contains about 1,000,000,000 erythrocytes. Each suspension contained the salivary glands from 9 to 177, usually 10-20 mosquitoes. Aliquots of the preparation were inoculated intravenously to White Rock or White Leghorn chicks, 2-7 days old, in groups of four or more.