Somerson N L, Kocka J P, Rose D, Del Giudice R A
Appl Environ Microbiol. 1982 Feb;43(2):412-7. doi: 10.1128/aem.43.2.412-417.1982.
The isolation of Mollicutes from food has not been reported. To isolate Mollicutes in the presence of high levels of unwanted bacteria, we first incubated fresh vegetables in liquid culture media containing lysozyme, ampicillin, and thallous acetate. Culture fluids were than separated from the vegetable samples, subjected to one freeze-thaw cycle, and passed through a filter of 0.4-micron porosity. Filtered samples were cultured in SP4 medium and in a conventional medium containing horse serum. With this procedure 21 acholeplasma isolations representing three species were obtained from endive, broccoli, and kale. Of 35 food samples tested, 11 were positive for acholeplasmas; acholeplasmas isolated from 6 of these samples were recovered only in SP4 medium. In seven single vegetable samples, two or more Acholeplasma spp. were isolated. A. laidlawii was isolated from all three vegetables and A. axanthum was found in broccoli and kale. Four isolates were serologically identified as A. oculi. Mycoplasma verecundum was the only Mycoplasma species recovered. Several isolates could not be typed serologically, as they reacted with antisera to both A. morum and A. hippikon. these isolates may include new Acholeplasma spp.
尚未见从食品中分离出柔膜菌纲微生物的报道。为了在存在大量不需要的细菌的情况下分离柔膜菌纲微生物，我们首先将新鲜蔬菜在含有溶菌酶、氨苄青霉素和醋酸铊的液体培养基中培养。然后将培养液与蔬菜样品分离，进行一次冻融循环，并通过孔径为0.4微米的滤膜。将过滤后的样品在SP4培养基和含有马血清的传统培养基中培养。通过该程序，从菊苣、西兰花和羽衣甘蓝中获得了代表三个物种的21株无胆甾原体分离株。在检测的35个食品样品中，11个无胆甾原体呈阳性；从其中6个样品中分离出的无胆甾原体仅在SP4培养基中生长。在7个单一蔬菜样品中，分离出了两种或更多种无胆甾原体属物种。从所有三种蔬菜中均分离出莱氏无胆甾原体，在西兰花和羽衣甘蓝中发现了黄色无胆甾原体。4株分离株经血清学鉴定为眼无胆甾原体。羞怯支原体是唯一分离到的支原体物种。有几株分离株无法进行血清学分型，因为它们与针对桑氏无胆甾原体和马无胆甾原体的抗血清均发生反应。这些分离株可能包括新的无胆甾原体属物种。
