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Aninvitro cultivation method for the predominant anaerobic eukaryotic microorganismEntodiniumcaudatum in the rumen of ruminants
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Qinhui XU1, Jinying HE1, Yujia WANG1, Cheng PENG1, Xiaoyue LU1, Zixian YANG1, Fahu YUAN1, Congjie HUA1, Jie XIONG2, Wei MIAO2, Jinmei FENG1, *
Acta Microbiologica Sinica | 2024, 64(7) : 2554 - 2565
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Acta Microbiologica Sinica | 2024, 64(7): 2554-2565
• Technology and Method •
Aninvitro cultivation method for the predominant anaerobic eukaryotic microorganismEntodiniumcaudatum in the rumen of ruminants
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Qinhui XU1, Jinying HE1, Yujia WANG1, Cheng PENG1, Xiaoyue LU1, Zixian YANG1, Fahu YUAN1, Congjie HUA1, Jie XIONG2, Wei MIAO2, Jinmei FENG1, *
Affiliations
  • 1 Department of Pathogenic Biology, School of Medicine, Jianghan University, Wuhan 430056, Hubei, China
  • 2 State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, Hubei, China
Published: 2024-07-04 doi: 10.13343/j.cnki.wsxb.20240004
Outline
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[Objective] To develop a stable and controllable method for isolating and cultivating the eukaryotic microorganismEntodiniumcaudatum from the rumen of ruminants, which would lay a foundation for the germplasm banking and provide sufficient experimental materials for researching the physiological function of rumen ciliates. [Methods] First, a rumen cannula was used to collect rumen fluid from cows in Wuhan, andE.caudatum was gradually enriched and isolated from the rumen fluid by micro-strainer filtration through different sizes of nylon mesh. Subsequently, the enrichedE.caudatum was cultured in anaerobic culture bottles loaded with the modified SP medium. After purification, a single culture of the strain was established. The species was identified by morphological observation and the phylogenetic analysis based on the 18S rRNA gene. Finally, the generation time of theE.caudatum was calculated based on the half-transfer cultivation method. [Results] A single culture ofE.caudatum was isolated from the rumen fluid of Holstein cows. The modified SP medium used in this study was composed of SP salt solution, supernatant of the rumen fluid without protozoa, cysteine hydrochloride, antibiotics, starch, and grass powder. In the modified SP medium, the obtained culture grew and proliferated steadily, reaching the highest density of 37 000 cells/mL on day 16 from the initial inoculation density of 320 cells/mL. Morphological features and molecular data indicated that the culture obtained in this study wasE.caudatum, namedE.caudatum strain WH. The generation time ofE.caudatum strain WH was 19.0 h. [Conclusion] This study has successfully developed aninvitro cultivation method forE.caudatum strain WH, a prevalent eukaryotic microorganism from the rumen of ruminants. This work lays a technical and theoretical basis for the germplasm banking and in-depth research on the physiological functions of rumen ciliates.

eukaryotic microorganism  /  rumen ciliate  /  Entodinium  /  modified SP medium  /  invitro cultivation  /  anaerobic cultivation
Qinhui XU, Jinying HE, Yujia WANG, Cheng PENG, Xiaoyue LU, Zixian YANG, Fahu YUAN, Congjie HUA, Jie XIONG, Wei MIAO, Jinmei FENG. Aninvitro cultivation method for the predominant anaerobic eukaryotic microorganismEntodiniumcaudatum in the rumen of ruminants[J]. Acta Microbiologica Sinica, 2024 , 64 (7) : 2554 -2565 . DOI: 10.13343/j.cnki.wsxb.20240004
Funding
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  • National Natural Science Foundation of China(32070420)
  • National Key Research and Development Program of China(2022YFF1001100)
Appendix
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Year 2024 volume 64 Issue 7
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Article Info
doi: 10.13343/j.cnki.wsxb.20240004
  • Receive Date：2024-01-03
  • Online Date：2026-03-19
  • Published：2024-07-04
Article Data
Affiliations
History
  • Received：2024-01-03
  • Accepted：2024-03-12
Funding
National Natural Science Foundation of China(32070420)
National Key Research and Development Program of China(2022YFF1001100)
Affiliations
    1 Department of Pathogenic Biology, School of Medicine, Jianghan University, Wuhan 430056, Hubei, China
    2 State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, Hubei, China

Corresponding:

*FENG Jinmei, E-mail:
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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