Iglesias L, Valero A, Adroher F J
Departamento de Parasitología, Facultad de Farmacia, Universidad de Granada, Spain.
Folia Parasitol (Praha). 1997;44(4):297-301.
Several culture media as well as some factors that may affect the in vitro development of the nematode Anisakis simplex Rudolphi, 1809 have been studied. After testing six media and four temperatures, the conditions for the in vitro culture selected were as follows: RPMI-1640 + 20% (v/v) heat-inactived fetal bovine serum or Meyer's M3 (without agar) media, at 37 degrees C, under 5% CO2 in air atmosphere, and renewal of the medium twice a week. The average survival rates of the larvae were significantly increased when the pH of the culture medium was increased (from 4.0 to 7.2) or decreased (from 7.2 to 4.0) after L3 to L4 moulting. The length of the larvae at the onset of culture affected the survival and moulting of themselves, but these were culture medium-dependent. On the other hand, we have observed that several L3 and L4 were attached, by means of a brown unknown substance apparently secreted by themselves, to the bottom of the substratum. Frequently, when a larva was spontaneously detached, a "cap" of the brown substance blocked, apparently, its mouth. The possible absorption of nutrients through the L3 larvae cuticle of A. simplex is discussed.
已经研究了几种培养基以及一些可能影响1809年鲁道夫氏简单异尖线虫体外发育的因素。在测试了六种培养基和四个温度后，选择的体外培养条件如下：RPMI - 1640 + 20%（v/v）热灭活胎牛血清或迈耶氏M3（无琼脂）培养基，在37摄氏度，空气气氛中5%二氧化碳条件下，每周更换两次培养基。当培养基的pH值在L3至L4蜕皮后升高（从4.0至7.2）或降低（从7.2至4.0）时，幼虫的平均存活率显著提高。培养开始时幼虫的长度影响其自身的存活和蜕皮，但这些都取决于培养基。另一方面，我们观察到一些L3和L4通过一种显然由它们自身分泌的褐色未知物质附着在培养基底部。通常，当幼虫自发脱离时，褐色物质的“帽”显然会堵塞其口部。讨论了简单异尖线虫L3幼虫角质层可能的营养吸收情况。
