微生物實驗：用豆粉浸液培養(yǎng)基培養(yǎng)花斑癬病菌
發(fā)布時間：
2022-12-27
作者：
摘要：分別用豆粉，豆油，豆粉浸液三種物質制成的固體培養(yǎng)基對花斑癬菌進行了分離培養(yǎng)的研究。結果表明：豆粉浸液固體培養(yǎng)基容易制備，易保存。清晰，菌落生長天數(shù)短，陽性率高等特點優(yōu)于豆油固體培養(yǎng)基，與豆粉固體培養(yǎng)基相比較，雖培養(yǎng)結果無差異，但前者易制備，易觀察。所以，我們認為豆粉浸液培養(yǎng)基較其它兩種有推廣使用價值。
我們采用豆粉、豆油、豆粉浸液制成的三種固體瓊脂培養(yǎng)基。培養(yǎng)了30例花斑堂皇壹良損鱗屑，并對其培養(yǎng)結果進行了比較，現(xiàn)將結果報告如下：
材料與方法：
1、取材對象：具有典幻皮損害。經(jīng)直接鏡檢真菌陽性的花斑癬患者30例，其中男性18例，女性12例，年齡10～50歲，平均年齡30歲。
2、培養(yǎng)基制備：①豆粉與豆油培養(yǎng)基。采用沙氏瓊脂，加熱溶化后，按4的比例分別加入豆粉和豆油，煮5～10min后。分裝，10磅10min滅菌，置斜面，備用。②豆粉浸液培養(yǎng)基：用蒸餾水1000m1加入30g豆粉，放冰箱過夜，次日取出，用100m1濾液加蛋白胨1g，瓊脂1。8g，葡萄糖4g的比例加入上述成分，加熱溶化。分裝，10磅10min滅菌，置斜面，備用。
3、取材及培養(yǎng)：取材前先將皮損區(qū)用75酒精消毒，然后用無菌鑷子輕輕刮取鱗屑。分別接種于上述三種培養(yǎng)基斜面上，置37℃溫箱中孵育，每天觀察菌落的生長情況，觀察時間為15天。
結果：1、菌落生長情況：1d后部分試管斜面上開始出現(xiàn)菌落。直徑約0.5～5mm大小。散在分布，逐漸長大融合成片。表面不光滑，有皺褶。初幾天為乳白色。以后漸變?yōu)楹稚＞湮⑼怀雠囵B(yǎng)基的表面。不向培養(yǎng)基內生產(chǎn)。1w內生長最快。以后減慢，10d后基本停止生長，15d后培養(yǎng)基漸變干。其菌落也皺縮，變?yōu)楹诤稚?。在固體培養(yǎng)基上，菌落比較旺盛。
2、陽性率：在接種的90支試管中去3支污染外，各培養(yǎng)基的陽性率。詳見表1三種培養(yǎng)基陽性率比較。
3、三種培養(yǎng)基生長天數(shù)：三種培養(yǎng)基從鱗屑接種開始。到肉眼可見菌落生長的最短，最長和平均天數(shù)結果見表2。
討論
1951年G0r0|0r首次采用橄欖油培養(yǎng)基對花斑癬菌培養(yǎng)成功以來。國外有相同結果的報道，近幾年國內有各種油類培養(yǎng)基代替橄欖油培養(yǎng)基進行實驗研究已取得良好效果n，但含油培養(yǎng)基易變質。保存時間短，油層易移層。存放不便，所以有人研究提出用豆粉固體培養(yǎng)基代替含油類。取得了令人滿意的效果。我們在使用豆粉固體培養(yǎng)基培養(yǎng)花斑癬菌中發(fā)現(xiàn)，豆粉固體培養(yǎng)基制備較困難。制備好的分上下兩層，豆粉都沉淀在下層，而花斑癬菌只在表面生長。就是說花斑癬菌生長利用的不是豆粉中的全部成份，而只一部分，所以我用豆粉制成浸液，并制成豆浸液固體培養(yǎng)基。并對豆粉、豆油、豆粉浸液三種培養(yǎng)基的結果進行比較。表1三種培養(yǎng)陽性率分別是96.66、i009／5、81.48。豆浸液與豆粉培養(yǎng)基無差異(p>0.05)，與豆油培養(yǎng)基有差異(p<0.05)，從三種培養(yǎng)基開始生長菌落的平均天數(shù)比較，豆浸液與豆粉培養(yǎng)基無差異(p>0.05)而與豆油培養(yǎng)基有差異(p<0.05)。實驗結果表明豆浸液培養(yǎng)基的陽性率高，培養(yǎng)速度快，易制備，易保存，完全可代替豆粉，豆油類培養(yǎng)基，值得推廣應用。
ABSTRACT: The isolation and culture of tinea versicolor were studied on the solid medium made of soybean powder, soybean oil and soybean powder extract. The results showed that the solid medium of soybean meal infusion was easy to prepare and easy to preserve. It is clear that the number of colony growth days is short and the positive rate is superior to the solid medium of soybean oil. Compared with the solid medium of soybean meal, the result is not different, but the former is easy to be prepared and easy to observe. Therefore, we believe that soybean meal infusion medium is more widely used than other two species.
Three solid agar media were made from soybean meal, soybean oil and soybean meal infusion. 30 cases of variegated flowers were cultured, and the results were compared. The results are as follows:
Materials and methods:
1. The object: the damage of the canonical skin. Thirty cases of tinea versicolor with positive fungi were examined by direct microscopy, including 18 males and 12 females, aged 10-50, with an average age of 30 years.
2. Preparation of medium: (1) soybean meal and soybean oil medium. After heating and dissolving, the soybean meal and soybean oil were added at 4 to 5 to 10min after heating. Separate, 10 pounds 10min sterilization, inclined plane, spare. (2) Soybean powder extract medium: add 30 g soybean powder with distilled water 1000 m1, put it in refrigerator overnight, take out the next day, add peptone 1 g with 100 M1 filter, agar 1. 8g, the ratio of glucose 4G was added to the above components and heated to melt. Separate, 10 pounds 10min sterilization, inclined plane, spare.
3, sampling and Cultivation: before taking the material, disinfect the lesion area with 75 alcohol, then gently scrape the scales with sterile forceps. The colonies were inoculated on the inclined surfaces of the three media and incubated in a 37 C incubator. The growth of the colonies was observed every day for 15 days.
Results: 1. Colony growth: colony appeared on some test tube slopes after 1D. The diameter is about 0.5 to 5mm. Scattered in the distribution, gradually grow into pieces. The surface is not smooth and has wrinkles. The first few days are milky white. It gradually became brown in the future. The colony is microprotruding the surface of the medium. It is not produced in the culture base. The fastest growth is in 1W. After slowing down, 10d stopped growing and 15d gradually dried. Its colony crinkled and became dark brown. On the solid medium, the colony is more vigorous.
2. Positive rate: the positive rate of each medium was 3 in the 90 test tubes. The positive rate of three kinds of medium is compared in Table 1.
1 Comparison of positive rates of three kinds of medium
3, three kinds of medium growth days: three medium starting from scale inoculation. To the naked eye, the shortest, longest and average days of colony growth can be seen in Table 2.
Table 2 Comparison of the growth days of three kinds of medium
discuss
In 1951, G0r0|0r first used olive oil medium for the successful cultivation of tinea versicolor. In recent years, there have been a variety of reports on the same results in foreign countries. In recent years, there have been a variety of oil media in place of the olive oil medium for experimental research, which has achieved good results n, but the oil containing medium is easy to deteriorate. The storage time is short, and the oil layer is easily moved. It is difficult to store. Therefore, some researchers have suggested that soybean meal solid medium should be used instead of oil. Satisfactory results have been achieved. We found that it was difficult to prepare soybean meal solid medium by using soybean meal solid medium to cultivate tinea versicolor. The upper and lower two layers were prepared, and soybean powder was deposited on the lower layer, while tinea versicolor was only growing on the surface. That is to say, the growth and utilization of tinea versicolor is not all the ingredients in the bean powder, but only a part, so I use the bean powder to make the soak and make the solid medium of the bean extract. The results of three media were compared with soybean meal, soybean oil and soybean meal. Table 1 the positive rates of three cultures were 96.66, I009 / 5 and 81.48 respectively. There was no difference between soybean extract and soybean meal medium (p>0.05), and there was a difference from soybean oil medium (p<0.05). The average days of growing colonies from three kinds of medium were compared. There was no difference between soybean extract and soybean meal medium (p>0.05), but there was a difference from soybean oil medium (p<0.05). The experimental results show that the culture medium of bean extract has high positive rate, fast culture speed, easy to be prepared, easy to preserve, and can completely replace bean powder and soybean oil medium. It is worth popularizing.
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