胰酪胨大豆瓊脂培養(yǎng)基平皿暴露時長研究
發(fā)布時間：
2022-12-24
作者：
微生物新型檢測培養(yǎng)基
胰酪胨大豆瓊脂培養(yǎng)基平皿暴露時長研究
研究對象：90mm TSA 平板，25ml和30ml兩種裝量，來自3個不同的廠家
研究策略：在單向?qū)恿髦蟹胖?.1小時和4.1小時，而后接菌培養(yǎng),真菌類20-25℃培養(yǎng)5天,細菌類30-35℃培養(yǎng)3天。對照平板沒有在層流中暴露，但是同時放入培養(yǎng)箱中培養(yǎng)。并有陰性對照。
細節(jié)：接菌方式為0.1ml菌液涂布；筆式水針車間D級區(qū)清洗室A級送風(fēng)四角及中部風(fēng)速，共5個點，各區(qū)域風(fēng)速均在0.36～0.54m/s，溫濕度符合日常要求范圍。
接種菌種列表：CMCC菌種
1. Bacillus subtilits 枯草芽孢桿菌
2. Candida albicans白色念珠菌
3. Staphylococcus aureus金黃色葡萄球菌
4. Aspergillus niger黑曲霉
5. Pseudomonas aeruginosa銅綠假單孢菌
回收率結(jié)果：
直徑為 90mm，裝量為 30mL 的 TSA 平皿在暴露 2.1h 和 4.1h 后，接種于表面的金黃色葡萄球菌、銅綠假單胞菌、枯草芽孢桿菌、白色念珠菌和黑曲霉的生長均符合要求；直徑為 90mm、裝量為 25mL 的 TSA 平皿在暴露 2.1h 后微生物的生長符合要求，但暴露 4.1h后，接種的黑曲霉生長情況不好。
我的討論：
本研究與【國外研究】在單向流下進行動態(tài)沉降菌測試時，平板失水對微生物生長性能影響比較有幾點不同：
1.國內(nèi)研究的接收標(biāo)準(zhǔn)是50%-200%，國外研究是70%
2.國內(nèi)研究的培養(yǎng)時間（3或者5天）要少于國外研究（5+2天）
3.沒有稱重，不清楚失水情況到底怎么樣
4.國內(nèi)研究比較不同廠家的產(chǎn)品，差別較大。甚至有些廠家的產(chǎn)品4.1h的回收率要好于2.1h，有些讓人費解。
5.國內(nèi)研究有些菌種接種量較少（小于10cfu），容易造成計算結(jié)果的波動較大。
6.沒有考慮環(huán)境分離菌株
本研究的價值在于讓人清楚的看到不同廠家的產(chǎn)品的巨大差異。結(jié)合國外研究中的結(jié)果可以看到，TSA平板的干粉原料不同以及干粉溶解滅菌后凝固的過程對TSA是否耐暴露是至關(guān)重要的，25ml到30ml的回收率的一點改變只是表面現(xiàn)象。但是因為本研究中沒有公布3家廠家的原料信息以及制備TSA平板的過程，所以無從考證。
本研究還可以看成是國內(nèi)TSA暴露時間驗證的一個樣板，如果你買的預(yù)制TSA廠家可以提供類似的報告也是不錯的。
做好的一次性胰酪大豆胨液體培養(yǎng)基，真心不容易。
（測試效果很好的日水牌胰酪胨大豆瓊脂培養(yǎng)基平皿圖，點圖查看詳情）
Study on exposure time of trypsin soy agar medium
Research object: 90mm TSA flat, 25ml and 30ml two loading capacity, from 3 different manufacturers.
Research strategy: placed in a one-way laminar flow for 2.1 hours and 4.1 hours, followed by bacteria culture, fungi at 20-25 degrees for 5 days, and bacteria for 30-35 days for 3 days. The control plate was not exposed in laminar flow, but was incubator in the incubator. And there was a negative control.
Details: the method of bacteria grafting is coated with 0.1ml bacteria, and the wind speed in the D level cleaning room of the pen type water needle workshop is four angles and 5 points in the middle. The wind speed in each area is 0.36 to 0.54m/s, and the temperature and humidity are in accordance with the daily requirements.
The list of inoculation bacteria: CMCC strain
Bacillus subtilis 1. Bacillus subtilits
2. Candida albicans Candida albicans
3. Staphylococcus aureus Staphylococcus aureus
4. Aspergillus Niger Aspergillus niger
5. Pseudomonas aeruginosa Pseudomonas aeruginosa
Recovery results:
The growth of Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans, Candida albicans and Aspergillus niger on the surface of the TSA plate with a diameter of 90mm and 30mL was all in accordance with the requirements. The growth character of the TSA flat with a diameter of 90mm and 25mL in 25mL was exposed to the growth character of the microbe after 2.1h. However, after exposure to 4.1h, the growth of Aspergillus niger was not good.
My discussion:
In this study, there were several differences in the effect of flat water loss on microbial growth performance under the unidirectional flow of dynamic sedimentation tests.
1. the acceptance standard of domestic research is 50%-200%, while foreign research is 70%
2. the incubation time of domestic research (3 or 5 days) is less than that of Foreign Studies (5+2 days).
3. no weighing. I don't know what the situation is.
4. domestic research compares the products of different manufacturers. Some manufacturers even have a better recovery rate of 4.1h than 2.1h, which is puzzling.
5. some domestic studies have less inoculation (less than 10cfu), which is likely to cause greater fluctuation in the calculation results.
6. no environmental isolates were considered
Overall, in this study, the TSA of 25ml capacity is almost completely annihilated, and 30ml loading is barely qualified, and the result is not optimistic.
The value of this research is to make people see clearly the huge difference of products from different manufacturers. According to the results from foreign studies, it is important to see whether the dry powder material of TSA plate and the solidification process after the dry powder dissolve and sterilization are essential to the resistance to exposure of TSA, and the change of the recovery rate of 25ml to 30ml is only a superficial phenomenon. However, because the material information of 3 manufacturers and the process of preparing TSA tablets were not published in this study, it is impossible to verify.
This study can also be seen as a model for the verification of TSA exposure time in China. If you buy a prefabricated TSA manufacturer, you can provide similar reports.
A good disposable trypsin soy peptone liquid medium is not easy at all.