Brown D F, Spaulding P L, Twedt R M
Appl Environ Microbiol. 1977 Jan;33(1):10-4. doi: 10.1128/aem.33.1.10-14.1977.
The enteropathogenicity of Vibrio parahaemolyticus was investigated by contrasting the effects of whole cells, cell fragments, cell-free preparations, and media constituents injected into rabbit ileal loops. Three of 20 cultures utilized were Kanagawa-negative strains from seawater and sea fish. The remaining 17 cultures included both Kanagawa-positive and -negative strains from Japanese victims of gastroenteritis. Broth culture filtrates concentrated 10-fold by dialysis against 30% Carbowax were unreactive, whereas lyophilized filtrates, regardless of Kanagawa type, as well as all sterile broth preparations containing greater than or equal to 5% NaCl gave positive reactions in the rabbit gut. In contrast, crude lysates derived from broth cultures of Kanagawa-positive strains caused loop dilatation; lysate supernatants were unreactive. Lysates of cells washed from brain heart infusion agar were more reactive than lysates from Trypticase soy agar-grown cells. When agar-grown cell lysates prepared by disruption in saline were dialyzed against distilled water, they were devoid of gut reactivity. Reactivity was restored in dialysands resuspended in saline and in dialysates concentrated 10-fold. The agar-grown cell lysates exhibited Kanagawa-type hemolysis. Our data support the conclusion that the rabbit loop reactivity observed with lyophilized, cell-free culture filtrates may result from excessively elevated NaCl concentrations, and that a toxic factor associated with large-cell particles may be dialyzable, depends on saline for expression, and resembles the Kanagawa hemolysin.
通过对比注射到兔回肠袢中的全细胞、细胞碎片、无细胞制剂和培养基成分的作用,研究了副溶血性弧菌的肠致病性。所使用的20株培养物中有3株是来自海水和海鱼的神奈川阴性菌株。其余17株培养物包括来自日本肠胃炎患者的神奈川阳性和阴性菌株。通过对30%聚乙二醇进行透析浓缩10倍的肉汤培养滤液无反应,而冻干滤液,无论神奈川类型如何,以及所有含≥5%氯化钠的无菌肉汤制剂在兔肠道中均呈阳性反应。相比之下,神奈川阳性菌株肉汤培养物的粗裂解物导致肠袢扩张;裂解物上清液无反应。从脑心浸液琼脂上洗下的细胞裂解物比胰蛋白胨大豆琼脂培养的细胞裂解物反应性更强。当通过在盐水中裂解制备的琼脂培养细胞裂解物用蒸馏水透析时,它们失去了肠道反应性。透析液重悬于盐水中以及浓缩10倍的透析液中反应性得以恢复。琼脂培养的细胞裂解物表现出神奈川型溶血。我们的数据支持以下结论:冻干的无细胞培养滤液在兔肠袢中观察到的反应性可能是由于氯化钠浓度过高所致,并且与大细胞颗粒相关的毒性因子可能是可透析的,其表达依赖于盐水,并且类似于神奈川溶血素。