Department of Animal and Dairy Sciences, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Bio-Tech Pharmacal Inc, Fayetteville, Arkansas, USA.
J Environ Sci Health B. 2021;56(3):272-281. doi: 10.1080/03601234.2020.1868236. Epub 2021 Jan 5.
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.
本研究旨在利用 16S rDNA 从商业玉米-豆粕家禽饲料中鉴定出典型的需氧分离株,对其进行分类,并将数据与之前对这些相同分离株的全基因组测序(WGS)分析进行比较。将 10 克商业玉米-豆粕家禽饲料在 100ml 胰蛋白酶大豆肉汤中匀浆 2 分钟,连续稀释,平板接种于胰蛋白酶大豆琼脂(TSA),在 37°C 下有氧培养 24 小时。随后,将 20 个独特的菌落划线分离于 TSA 平板上,在 37°C 下有氧培养 24 小时。重复此过程三次,以纯化分离株,直到获得 11 个形态上不同的菌落。使用 Qiagen 的 DNeasey® Blood and Tissue Kit 提取 DNA。使用 Illumina MiSeq 靶向 16S rRNA V4 区,并通过 QIIME2-2020.2 进行分析。生成了 Alpha 多样性和 Beta 多样性指标,并在 Qiime2-2020.2 中使用 Silva 对齐分类群。在 11 个不同的菌落中鉴定出 25 个不同的属。由于 16S rDNA 鉴定可以了解病原体与生态系统中微生物生境的关联,因此该信息可能提供了一种建立和描述与家禽饲料相关的卫生指示微生物的潜在方法。