Phenotypic characterization of Histoplasma species

Evaluation of extracellular proteolytic activity: To evaluate extracellular proteolytic activity in different species of Histoplasma, we grew the 27 Histoplasma strains (Table S1) in HMM plates supplemented with 1.5% skim milk. Strains with a proteolytic activity show a clear halo around their yeast colonies.Optical density and growth curves of Histoplasma yeast cultures: We mixed 600 μl of liquid culture with 300 μl of 3M NaOH in a plastic cuvette, which was covered with Parafilm and vortexed for 10 seconds to separate yeast clumps and measure optical density (OD) in a GENESYS 10vis spectrophotometer (Thermo Spectronic) starting at day 0, and at every 24 hours after that until day 11.Yeast Area: We mixed 10 μl from a yeast culture that had large yeast clumps removed with 10 μl of Lactophenol Cotton Blue on a glass slide. Differential interference contrast (DIC) images were obtained using 100X/1.4 Oil UPlan S Apo PSF quality objective on an Olympus BX-61 microscope and collected using a QImaging RETIGA 4000R color camera and Volocity 6.3 acquisition software.Description of the data and file structureWe are including four files: Three of them are experimental observations, and one of them is the code required to analyze the data.halo_radius_miss.csv: ratio sizes of H. mississipiense cultures in .5% skim milk media. The variable halo is in cms and reflects the ratio of the lysis halo. Isolate refers to the name of the Histoplasma strain.od_cells_histo.csv: rates of growth of different species of Histoplasma in liquid media. Strain refers to the name of the Histoplasma strain. OD refers to the turbidity of the culture (reported in Absorbance Units). Cells is the number of cells in the culture.Cell_Size_df.csv: cell size for different species of Histoplasma. Line refers to the name of the Histoplasma strain. Replicate refers to the number of the experimental batch. Area is the ellipse area of the cell (in micrometers^2).histoplasma_morphology_code.R: Analytical code to generate linear models.胞外蛋白水解活性检测：为检测不同组织胞浆菌（Histoplasma）物种的胞外蛋白水解活性，我们将27株组织胞浆菌（表S1）接种于添加1.5%脱脂乳的HMM平板中培养。具备蛋白水解活性的菌株，其酵母菌落周围会形成清晰的透明晕圈。 组织胞浆菌酵母菌培养物的光密度与生长曲线测定：我们将600 μl液体培养物与300 μl 3M氢氧化钠（NaOH）置于塑料比色皿中混合，用帕拉菲膜（Parafilm）密封后涡旋振荡10秒以打散酵母菌团，随后从第0天开始，每隔24小时使用GENESYS 10vis型分光光度计（Thermo Spectronic）测量光密度（Optical Density, OD），直至第11天。 酵母菌面积测定：我们将已去除大菌团的酵母菌培养物中的10 μl与10 μl乳酸酚棉蓝染液（Lactophenol Cotton Blue）在载玻片上混合。使用奥林巴斯BX-61型显微镜搭配100X/1.4 Oil UPlan S Apo PSF物镜获取微分干涉差（Differential Interference Contrast, DIC）图像，并通过QImaging RETIGA 4000R彩色相机与Volocity 6.3采集软件完成图像采集。 数据与文件结构说明：本数据集包含4个文件，其中3个为实验观测数据，1个为数据分析所需代码。 halo_radius_miss.csv：密西西比组织胞浆菌（H. mississipiense）在0.5%脱脂乳培养基中的菌落晕圈比例数据。变量halo的单位为厘米（cm），代表裂解晕圈的比例；Isolate字段表示组织胞浆菌菌株的名称。 od_cells_histo.csv：不同组织胞浆菌物种在液体培养基中的生长速率数据。Strain字段表示组织胞浆菌菌株的名称；OD表示培养物的浊度，单位为吸光度单位（Absorbance Units）；Cells字段表示培养物中的细胞总数。 Cell_Size_df.csv：不同组织胞浆菌物种的细胞尺寸数据。Line字段表示组织胞浆菌菌株的名称；Replicate字段为实验批次编号；Area字段表示细胞的椭圆面积，单位为平方微米（μm²）。 histoplasma_morphology_code.R：用于构建线性模型的分析代码。