Liu Chang, Cheng Jin, Xie Yunong, Ouyang Kehui, Qu Mingren, Pan Ke, Qiu Qinghua
Jiangxi Province Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, China.
Vet Sci. 2025 Mar 3;12(3):234. doi: 10.3390/vetsci12030234.
The aim of this study was to investigate the effects of storage temperature and preservation time on the microbial diversity and community composition of rumen fluid. Rumen fluid samples were collected from six Hu sheep fed on a high-forage diet and stored at -80 °C and -20 °C for intervals of 0, 7, 14, 30, 60, 120, and 240 days. DNA was extracted at each time point for 16S rRNA gene sequencing to evaluate the rumen microbial diversity and community composition. The results showed that storage temperature affected only the relative abundance of Proteobacteria, with no substantial impact on alpha-diversity or other microbial groups ( > 0.05), and no significant interaction effects were observed between storage temperature and preservation time ( > 0.05). Alpha-diversity indices such as Chao1, observed species, and PD whole tree showed dynamic changes after 7 days of storage, while the relative abundances of Verrucomicrobiota and , as well as the energy metabolism metabolic pathway, exhibited significant alterations after 14 days of storage ( < 0.05). Notably, Patescibacteria, , and abundances demonstrated significant changes after 240 days of storage ( < 0.05). Both principal coordinates analysis (PCoA) and non-metric multidimensional scaling (NMDS) showed distinct overlaps. This study suggests that storing rumen fluid at -80 °C and -20 °C does not influence rumen microbial diversity and community composition, whereas the storage time significantly impacts these factors, with most differences emerging after 14 days of preservation. Consequently, it is advised that the analysis of microbial diversity and community composition in rumen fluid samples be conducted within 14 days post-collection.
本研究的目的是调查储存温度和保存时间对瘤胃液微生物多样性和群落组成的影响。从六只采食高粗饲料日粮的湖羊采集瘤胃液样本，并在-80°C和-20°C下分别储存0、7、14、30、60、120和240天。在每个时间点提取DNA进行16S rRNA基因测序，以评估瘤胃微生物多样性和群落组成。结果表明，储存温度仅影响变形菌门的相对丰度，对α多样性或其他微生物类群没有实质性影响（>0.05），且未观察到储存温度和保存时间之间的显著交互作用（>0.05）。Chao1、观测物种数和PD全树等α多样性指数在储存7天后呈现动态变化，而疣微菌门的相对丰度以及能量代谢代谢途径在储存14天后出现显著变化（<0.05）。值得注意的是，在储存240天后，Patescibacteria、以及的丰度出现显著变化（<0.05）。主坐标分析（PCoA）和非度量多维尺度分析（NMDS）均显示出明显的重叠。本研究表明，在-80°C和-20°C下储存瘤胃液不会影响瘤胃微生物多样性和群落组成，而储存时间会显著影响这些因素，大多数差异在保存14天后出现。因此，建议在采集瘤胃液样本后14天内进行微生物多样性和群落组成分析。
