Schmitt-Slomska J, Caravano R, Anoal M, Gay B, Roux J
Ann Microbiol (Paris). 1981 May-Jun;132(3):253-65.
Previous attempts to obtain in vitro wall-deficient stable L-forms of various strains of Brucella have failed because the obtained spheroplasts revert quickly to bacterial form. Here, we report the isolation of L-forms from mice infected with a B. suis strain type 1 and treated with penicillin. In defined experimental conditions, L-type microcolonies associated with tissue debris were observed in primary spleen cultures, even on antibiotic free media. After several transfers on penicillin-containing medium. typical, tissue-free L colonies were obtained. At first, when cultivated on antibiotic-free medium, these colonies reverted to the bacterial form (identified as B suis, biotype 1). Later, after approximately fifteen transfers on penicillin-supplemented medium, they no longer reverted even after several subcultures on antibiotic-free medium. The L-forms' ultrastructural features included many giant empty bodies, considerable variation in size, shape and density of the wall-deficient cells, and many multilayered membranes. The stabilized L-forms were propagated in vitro and inoculated into mice, and then recovered from their spleens as tissue associated L-microcolonies. An occasional in vivo revertant was identified as B. suis, biotype 1. These data provide one possible explanation for earlier failures to detect the presence of atypical bacteria in clinical or experimental Brucella infections.
此前，为获得布鲁氏菌各种菌株的体外细胞壁缺陷稳定L型菌的尝试均告失败，因为所获得的原生质体很快就会回复到细菌形态。在此，我们报告了从感染1型猪布鲁氏菌并经青霉素处理的小鼠中分离出L型菌。在特定的实验条件下，即使在无抗生素培养基上，在原代脾脏培养物中也观察到了与组织碎片相关的L型小菌落。在含青霉素的培养基上多次传代后，获得了典型的、无组织的L菌落。起初，当在无抗生素培养基上培养时，这些菌落回复到细菌形态（鉴定为猪布鲁氏菌生物1型）。后来，在补充青霉素的培养基上大约传代15次后，即使在无抗生素培养基上多次传代，它们也不再回复。L型菌的超微结构特征包括许多巨大的空泡、细胞壁缺陷细胞在大小、形状和密度上有相当大的差异，以及许多多层膜。稳定的L型菌在体外繁殖并接种到小鼠体内，然后从它们的脾脏中作为与组织相关的L型小菌落回收。偶尔在体内出现的回复菌被鉴定为猪布鲁氏菌生物1型。这些数据为早期在临床或实验性布鲁氏菌感染中未能检测到非典型细菌的存在提供了一种可能的解释。
