芽胞杆菌菌体浓度的快速检测方法.docVIP

芽胞杆菌快速 刘国红1，2，林营志1，林乃铨1，刘波1 （1 福建省农业科学院农业生物资源所 350003，2 福建农林大学 350002） 摘要： 关键词 Study on speedy method of genus Bacillus biological characteristics Abstract：Bacillus is a type of rod-shaped cells, producing spores of the bacteria groupsmedicine area. So, it was important to study biological characteristics of genus Bacillus, which playing a role in application. Generally, the traditional statistical method was the concentration of Bacillus coated plates or diluted blood count plate count, which were time-consuming, and results were impacted by the subjective largely. In this paper, we took Bacillus cereus cultured in different pH as example, measured the absorbance of different pH by spectrophotometry, water and culture medium as a blank control respectively, and its corresponding concentration of Bacillus cereus were compared. Last, by DPS software, we analyzed the results finding that there was a very significant correlation between the OD values ??measured and the bacterial concentration. The results demonstrated that Spectrophotometric detection can replace the concentration of Bacillus cereus expressing the biological characteristics of Bacillus, and this method is simple, fast and reliable. Keywords: Bacillus; biological characteristics; OD value; 前言 通常采用显微镜计数法测定菌浓度，该方法费时费力，且容易让观察者眼睛过度疲劳，从而产生不必要的主观性误差。本文针对培养过程中菌浓度的传统方法进行改进，对培养过程中的进行了浓度与培养液吸光度的相关性分析。研究菌浓度与吸光度的关系，以便利用该相关性对菌的生长情况进行简便、快速的测定。 材料与方法 材料 试验用水为超纯水，紫外可见分光光度计(UV-2550，220V)，恒温培养振荡器（ZHWY-2102C，220V），显微镜，高速离心机。 试验方法 菌液的制备：以不同pH值NA液体培养基培养24 h的菌液为测试样，然后将菌液稀释为一系列不同浓度的菌液，三个平行样品，即三个重复。菌液浓度测定方法：采用血球计数板法（25×16），在光学显微镜下计数，三次计数，取均值。菌液吸光度的测定：分别以水和培养基作为空白对照，测定菌液的吸光度（OD值）。上清液的处理：培养24 h的菌液离心3000 r.min-1 20min后，取上清液。 菌浓度与吸光度的相关性分析 分别测定菌液和对应上清液在各波长下的吸光度值，利用DPS软件进行数据处理，拟合菌液浓度-吸光度线性关系。 2.1藻培养液的吸光特性 分别对芽胞杆菌菌液和上清液进行光谱扫描，如图1和图2所示由图1可以看出，菌液可见光波长范围内没有很明显的吸收峰，在可见波长发范围内随波长减小吸光度呈上升趋势。而OD600是追踪液体培养物中微生物生长的标准方法。故选取在波长600 nm测定芽胞杆菌菌液的吸光度。图2为菌液上清液的光谱扫描图，很明显地发现在整个可见波长范围内没有吸收峰，且吸光度值几乎接近0点，由此说明上清液对菌液的吸光度值几乎没有影响，可忽略不计。 图 1 芽胞杆菌菌液的光谱扫描图 图