PMID- 24118832 OWN - NLM STAT- MEDLINE DCOM- 20160422 LR - 20181202 IS - 1365-2672 (Electronic) IS - 1364-5072 (Linking) VI - 116 IP - 1 DP - 2014 Jan TI - Fluorescence in situ hybridization probing of protozoal Entodinium spp. and their methanogenic colonizers in the rumen of cattle fed alfalfa hay or triticale straw. PG - 14-22 LID - 10.1111/jam.12356 [doi] AB - AIMS: To develop and test a fluorescence in situ hybridization (FISH) based technique and to identify and quantify simultaneously those methanogenic populations colonizing Entodinium spp. in the rumen of cows fed different forages. METHODS AND RESULTS: New FISH probes targeting protozoal Entodinium spp. were designed and used together with FISH probes for methanogens in the cow rumen. The composition and relative abundance of methanogenic populations colonizing Entodinium simplex-, E. caudaum- and Entodinium furca-related populations were similar. Methanogens including Methanobrevibacter thaueri, Methanobrevibacter millerae and Methanobrevibacter smithii, and members of Methanomicrobium and Methanosphaera were generally the predominant colonizers of protozoa, regardless of the forage fed to cattle. Individual animals appeared to differ in which ruminal methanogenic populations colonized each of the individual Entodinium spp. CONCLUSIONS: Simultaneous FISH probing is shown here to be a reliable and effective approach to investigate the dynamics of symbiotic relationships between ruminal protozoa and methanogens at a single cell level. Phylogenetically closely related Entodinium spp. were colonized by similar methanogenic populations regardless of the forage fed. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of the methanogenic archaeal populations that specifically colonize Entodinium spp. as identified using simultaneous FISH probing. CI - Her Majesty the Queen in Right of Canada (c) 2013 Society for Applied Microbiology Reproduced with the permission of the Minister of Agriculture and Agri-Food Canada. FAU - Xia, Y AU - Xia Y AD - Lethbridge Research Centre, Agriculture and Agri-Food Canada, Lethbridge, AB, Canada; Department of Biological Science and Technology, Kunming University, Kunming, China. FAU - Kong, Y H AU - Kong YH FAU - Seviour, R AU - Seviour R FAU - Forster, R J AU - Forster RJ FAU - Kisidayova, S AU - Kisidayova S FAU - McAllister, T A AU - McAllister TA LA - eng PT - Journal Article DEP - 20131024 PL - England TA - J Appl Microbiol JT - Journal of applied microbiology JID - 9706280 RN - 0 (RNA, Ribosomal, 16S) SB - IM MH - Animals MH - Cattle MH - Euryarchaeota MH - In Situ Hybridization, Fluorescence MH - *Medicago sativa MH - RNA, Ribosomal, 16S MH - *Rumen/parasitology MH - Triticale OTO - NOTNLM OT - Entodinium OT - fluorescence in situ hybridization OT - methanogens OT - rumen ecology OT - symbiosis EDAT- 2013/10/15 06:00 MHDA- 2016/04/23 06:00 CRDT- 2013/10/15 06:00 PHST- 2013/02/08 00:00 [received] PHST- 2013/07/12 00:00 [revised] PHST- 2013/09/23 00:00 [accepted] PHST- 2013/10/15 06:00 [entrez] PHST- 2013/10/15 06:00 [pubmed] PHST- 2016/04/23 06:00 [medline] AID - 10.1111/jam.12356 [doi] PST - ppublish SO - J Appl Microbiol. 2014 Jan;116(1):14-22. doi: 10.1111/jam.12356. Epub 2013 Oct 24.