Abstract

The study’s objective was to identify typical aerobic isolates from commercial, corn-soybean meal poultry diets utilizing 16S rDNA, assign them their corresponding taxonomy, and compare the data with the previously published WGS analysis of these same isolates. Ten grams of a commercial corn-soybean meal poultry diet was homogenized in 100 mL of tryptic soy broth for 2 min, serially diluted, plated onto tryptic soy agar (TSA), and incubated aerobically for 24 h at 37 °C. Subsequently, 20 unique colonies were streaked for isolation on TSA and incubated aerobically for 24 h at 37 °C. This process was repeated three consecutive times for purification of isolates until only 11 morphologically distinct colonies were obtained. DNA was extracted using Qiagen’s DNeasey® Blood and Tissue Kit. The 16S rRNA V4 region was targeted using an Illumina MiSeq and analyzed via QIIME2-2020.2. Alpha diversity and Beta diversity metrics were generated, and taxa were aligned using Silva in Qiime2-2020.2. Twenty-five distinct genera were identified within the 11 different colonies. Because 16S rDNA identification can provide an understanding of pathogen associations and microbial niches within an ecosystem, the information may present a potential method to establish and characterize the hygienic indicator microorganisms associated with poultry feed.

 抽象的

该研究的目的是利用 16S rDNA 从商业玉米豆粕家禽日粮中识别出典型的需氧菌株，为它们分配相应的分类，并将数据与之前发表的这些相同菌株的 WGS 分析数据进行比较。将 10 克商业玉米豆粕家禽饲料在 100 mL 胰蛋白酶大豆肉汤中均质化 2 分钟，连续稀释，铺在胰蛋白酶大豆琼脂 (TSA) 上，并在 37 °C 下有氧孵育 24 小时。随后，在 TSA 上划线分离 20 个独特的菌落，并在 37°C 有氧条件下孵育 24 小时。该过程连续重复3次以纯化分离株，直到仅获得11个形态不同的菌落。使用 Qiagen 的 DNasey® 血液和组织试剂盒提取 DNA。使用 Illumina MiSeq 靶向 16S rRNA V4 区域，并通过 QIIME2-2020.2 进行分析。生成了 Alpha 多样性和 Beta 多样性指标，并使用 Qiime2-2020.2 中的 Silva 对齐了分类单元。在 11 个不同的菌落中鉴定出 25 个不同的属。由于 16S rDNA 鉴定可以提供对生态系统内病原体关联和微生物生态位的了解，因此该信息可能提供一种潜在的方法来建立和表征与家禽饲料相关的卫生指示微生物。