胎牛血清脑心浸液琼脂培养基在马尔尼菲青霉酵母相转相中的应用

The Application of Fetal Calf Serum Brain Heart Infusion Ager in Yeast Phase Transition of Penicillium Marneffei

目的探讨胎牛血清脑心浸液琼脂培养基在马尔尼菲青霉由菌丝相向酵母相转相中的应用价值,以及两种不同来源马尔尼菲青霉(即临床人感染株及广西野生银星竹鼠寄生株)在该培养基中转相程度的异同。方法取临床人感染株18株及竹鼠寄生株13株分别接种在已配制好的胎牛血清脑心浸液琼脂培养基(实验组)及普通脑心浸液琼脂培养基(对照组)上并置于37°C恒温恒湿箱中培养7d,再连续转种2次后,取菌落均匀涂片并予乳酸酚棉兰染色,在高倍显微镜下观察并计算单个酵母样细胞和关节孢子占该镜下所有细胞的百分比(即转化率),连续取卫星位6个镜下视野百分值的平均值作为该菌最终转化率。结果马尔尼菲青霉临床人感染株酵母相下单个酵母样细胞和关节孢子占每镜下所有细胞百分比为(75.08±3.32)%,对照组为(54.09±4.92)%;竹鼠寄生株实验组为(73.36±2.64)%,对照组为(52.37±2.57)%;两种不同来源马尔尼菲青霉酵母相下实验组与对照组比较差异均有统计学意义(P<0.05);临床人感染株的实验组和对照组与竹鼠寄生株相应组别比较差异均无统计学意义(P>0.05)。结论胎牛血清脑心浸液琼脂培养基在马尔尼菲青霉酵母相转相中有较高的应用价值,值得推广;两种不同来源马尔尼菲青霉酵母相在胎牛血清脑心浸液琼脂培养基中转相程度无区别。

Objective To investigate the application of fetal calf serum （FBS） brain heart infusion ager in the switching to yeast cells from hyphae of Penicillium marneffei （PM）, and the different degree of switching in two different sources of isolates from clinical patients and wild-Rhizomys of Guangxi. Methods We selected 18 strains of clinical isolates and 13 strains wild-Rhizomys＇s randomly, and cultivated them in the FBS brain heart infusion ager（ treating group） and the general BHI ager（ control group） respectively. Then we put the ready medium into incubator at 37℃ ,and kept for 7 days. It was found that these fungi converted to yeasts colonies gradually. After that we cuhivated little these yeasts colonies in the above two sorts of ager in the same way twice again. Since the lactophenol cotton blue would be used in the staining, we observed and calculated all single yeast-like cells and arthroconidias share the percentage of all cells in one endoscopic vision（ that＇s conversion rate ）. Finally, the average level of 6 different visions aroundly would be looked as the end percentile. Results The conversion rate of treating group of isolates from clinical patients was （ 75.08±3.32 ） % , and control group was （ 54.09± 4.92 ） % ; wild-Rhizomys＇s treating group was （ 73.36±2.64 ） % and control group was （52.37±2.57 ） %. It was found that there were more yeasts cells generated in treating group than in control one （ P 〈 0.05 ） regardless of the isolates from clinical patients or wild-Rhizomys. However, the results showed no difference in conversion rates between the two treating groups or control groups of the two different sources of isolates （ P 〉 0. 05 ）. Conclusion The FBS brain heart infusion agar has high application value and worth promoting. The results also showed that no difference in the degree of yeasts conversion of the two different sources of PM isolates.