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Karlheinz Schleifer - One of the best experts on this subject based on the ideXlab platform.
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identification of rumen Ciliates using small subunit ribosomal rna 18s rrna targeted oligonucleotide probes and fluorescence in situ hybridization fish
Journal of Eukaryotic Microbiology, 2005Co-Authors: J. Fried, Nadine A Thomas, Johannes H P Hackstein, Neil R. Mcewan, S. Kišidayová, Jamie C Newbold, Wolfgang Ludwig, Tadeusz Michalowski, S Weber, Karlheinz SchleiferAbstract:Up to half of the biomass in the rumen can be represented by Ciliates, which play an important role in the digestion processes of their hosts. In the literature, very little information can be found on determination of the diversity of complex rumen ciliate communities. The identification of these fast moving protists is mainly based on live observations and comparisons of their highly variable cell morphologies. It makes accurate identification and quantification of rumen Ciliates very difficult if not impossible. The development of fluorescence in situ hybridization (FISH), established as a technique to identify prokaryotes and eukaryotes using ribosomal RNA-targeted fluorescently labeled oligonucleotide probes, is a promising approach to identify and investigate rumen Ciliates. The present study, part of CIMES (Ciliates as monitors for environmental safety), a project sponsored by the European Commission, shows the problems of applying FISH on rumen Ciliates and how to solve them. Tests resulted in a new protocol, which recommends para-formaldehyde and formaldehyde at 1–2% final concentration to preserve the Ciliates before applying FISH. Furthermore, seven new oligonucleotide probes could be developed and successfully be tested to identify different rumen ciliate taxa of the order Entodiniomorphida (class Litostomatea) by applying FISH. It is also shown, how FISH together with confocal laser scanning microscopy can improve analyses of ciliate cell morphologies. Thanks to Peter Pristas, Peter Javorsky, Ralf Einspanier, Susanne Ulbrich (providing rumen samples), Seung Yeo Moon-van der Staay, Georg van der Staay (providing unpublished 18S-rDNA sequences), the European Commission (financial support, project QLK3-CT-2002-02151).
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Identification of rumen Ciliates using small subunit ribosomal RNA (18S‐rRNA)‐targeted oligonucleotide probes and fluorescence in situ hybridization (FISH)
Journal of Eukaryotic Microbiology, 2005Co-Authors: J. Fried, Nadine A Thomas, Johannes H P Hackstein, Tadeusz Michałowski, Neil R. Mcewan, S. Kišidayová, Jamie C Newbold, Wolfgang Ludwig, S Weber, Karlheinz SchleiferAbstract:Up to half of the biomass in the rumen can be represented by Ciliates, which play an important role in the digestion processes of their hosts. In the literature, very little information can be found on determination of the diversity of complex rumen ciliate communities. The identification of these fast moving protists is mainly based on live observations and comparisons of their highly variable cell morphologies. It makes accurate identification and quantification of rumen Ciliates very difficult if not impossible. The development of fluorescence in situ hybridization (FISH), established as a technique to identify prokaryotes and eukaryotes using ribosomal RNA-targeted fluorescently labeled oligonucleotide probes, is a promising approach to identify and investigate rumen Ciliates. The present study, part of CIMES (Ciliates as monitors for environmental safety), a project sponsored by the European Commission, shows the problems of applying FISH on rumen Ciliates and how to solve them. Tests resulted in a new protocol, which recommends para-formaldehyde and formaldehyde at 1–2% final concentration to preserve the Ciliates before applying FISH. Furthermore, seven new oligonucleotide probes could be developed and successfully be tested to identify different rumen ciliate taxa of the order Entodiniomorphida (class Litostomatea) by applying FISH. It is also shown, how FISH together with confocal laser scanning microscopy can improve analyses of ciliate cell morphologies. Thanks to Peter Pristas, Peter Javorsky, Ralf Einspanier, Susanne Ulbrich (providing rumen samples), Seung Yeo Moon-van der Staay, Georg van der Staay (providing unpublished 18S-rDNA sequences), the European Commission (financial support, project QLK3-CT-2002-02151).
Neil R. Mcewan - One of the best experts on this subject based on the ideXlab platform.
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The symbiotic intestinal Ciliates and the evolution of their hosts
European journal of protistology, 2014Co-Authors: Seung Yeo Moon-van Der Staay, Tadeusz Michałowski, Neil R. Mcewan, Peter Javorsky, Svetlana Kisidayova, Georg W.m. Van Der Staay, Jean-pierre Jouany, Peter Pristas, Zora Váradyová, C. Jamie NewboldAbstract:The evolution of sophisticated differentiations of the gastro-intestinal tract enabled herbivorous mammals to digest dietary cellulose and hemicellulose with the aid of a complex anaerobic microbiota. Distinctive symbiotic Ciliates, which are unique to this habitat, are the largest representatives of this microbial community. Analyses of a total of 484 different 18S rRNA genes show that extremely complex, but related ciliate communities can occur in the rumen of cattle, sheep, goats and red deer (301 sequences). The communities in the hindgut of equids (Equus caballus, Equus quagga), and elephants (Elephas maximus, Loxodonta africanus; 162 sequences), which are clearly distinct from the ruminant ciliate biota, exhibit a much higher diversity than anticipated on the basis of their morphology. All these Ciliates from the gastro-intestinal tract constitute a monophyletic group, which consists of two major taxa, i.e. Vestibuliferida and Entodiniomorphida. The Ciliates from the evolutionarily older hindgut fermenters exhibit a clustering that is specific for higher taxa of their hosts, as extant species of horse and zebra on the one hand, and Africa and Indian elephant on the other hand, share related Ciliates. The evolutionary younger ruminants altogether share the various entodiniomorphs and the vestibuliferids from ruminants.
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The symbiotic intestinal Ciliates and the evolution of their hosts
European Journal of Protistology, 2014Co-Authors: Seung Yeo Moon-van Der Staay, Neil R. Mcewan, Tadeusz Michalowski, Peter Javorsky, J Pierre Jouany, Svetlana Kisidayova, Georg W.m. Van Der Staay, Peter Pristas, Zora Váradyová, C. Jamie NewboldAbstract:The evolution of sophisticated differentiations of the gastro-intestinal tract enabled herbivorous mammals to digest dietary cellulose and hemicellulose with the aid of a complex anaerobic microbiota. Distinctive symbiotic Ciliates, which are unique to this habitat, are the largest representatives of this microbial community. Analyses of a total of 484 different 18S rRNA genes show that extremely complex, but related ciliate communities can occur in the rumen of cattle, sheep, goats and red deer (301 sequences). The communities in the hindgut of equids (Equus caballus, Equus quagga), and elephants (Elephas maximus, Loxodonta africanus; 162 sequences), which are clearly distinct from the ruminant ciliate biota, exhibit a much higher diversity than anticipated on the basis of their morphology. All these Ciliates from the gastro-intestinal tract constitute a monophyletic group, which consists of two major taxa, i.e. Vestibuliferida and Entodiniomorphida. The Ciliates from the evolutionarily older hindgut fermenters exhibit a clustering that is specific for higher taxa of their hosts, as extant species of horse and zebra on the one hand, and Africa and Indian elephant on the other hand, share related Ciliates. The evolutionary younger ruminants altogether share the various entodiniomorphs and the vestibuliferids from ruminants. (C) 2014 Elsevier GmbH. All rights reserved.
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The Onset of Ciliate Populations in Newborn Foals
Acta Protozoologica, 2010Co-Authors: Catherine Elizabeth Egan, Timothy J. Snelling, Neil R. McewanAbstract:The time of onset of survival of ciliate protozoa in the equine hindgut in new born foals was investigated. Daily faecal samples were collected from 6 new-born foals and studied under a microscope for examples of Ciliates within the samples. The results of this study show that Ciliates are first seen in faecal samples from the foal on day 5 post partum although these appeared to be voided and were assumed to be non-viable. However, by the following day the Ciliates collected seen in the faecal samples appeared to be intact and were assumed to be viable. This observation is 5 days earlier than Ciliates have previously been observed in faeces collected from the digestive tract of newborn foals.
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horizontal gene transfer from bacteria to rumen Ciliates indicates adaptation to their anaerobic carbohydrates rich environment
BMC Genomics, 2006Co-Authors: Guenola Ricard, Neil R. Mcewan, Didier Macheboeuf, Bas E Dutilh, J P Jouany, Makoto Mitsumori, Freda M Mcintosh, T Michalowski, Takafumi Nagamine, Nancy NelsonAbstract:The horizontal transfer of expressed genes from Bacteria into Ciliates which live in close contact with each other in the rumen (the foregut of ruminants) was studied using ciliate Expressed Sequence Tags (ESTs). More than 4000 ESTs were sequenced from representatives of the two major groups of rumen Cilates: the order Entodiniomorphida (Entodinium simplex, Entodinium caudatum, Eudiplodinium maggii, Metadinium medium, Diploplastron affine, Polyplastron multivesiculatum and Epidinium ecaudatum) and the order Vestibuliferida, previously called Holotricha (Isotricha prostoma, Isotricha intestinalis and Dasytricha ruminantium). A comparison of the sequences with the completely sequenced genomes of Eukaryotes and Prokaryotes, followed by large-scale construction and analysis of phylogenies, identified 148 ciliate genes that specifically cluster with genes from the Bacteria and Archaea. The phylogenetic clustering with bacterial genes, coupled with the absence of close relatives of these genes in the Ciliate Tetrahymena thermophila, indicates that they have been acquired via Horizontal Gene Transfer (HGT) after the colonization of the gut by the rumen Ciliates. Among the HGT candidates, we found an over-representation (>75%) of genes involved in metabolism, specifically in the catabolism of complex carbohydrates, a rich food source in the rumen. We propose that the acquisition of these genes has greatly facilitated the Ciliates' colonization of the rumen providing evidence for the role of HGT in the adaptation to new niches.
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identification of rumen Ciliates using small subunit ribosomal rna 18s rrna targeted oligonucleotide probes and fluorescence in situ hybridization fish
Journal of Eukaryotic Microbiology, 2005Co-Authors: J. Fried, Nadine A Thomas, Johannes H P Hackstein, Neil R. Mcewan, S. Kišidayová, Jamie C Newbold, Wolfgang Ludwig, Tadeusz Michalowski, S Weber, Karlheinz SchleiferAbstract:Up to half of the biomass in the rumen can be represented by Ciliates, which play an important role in the digestion processes of their hosts. In the literature, very little information can be found on determination of the diversity of complex rumen ciliate communities. The identification of these fast moving protists is mainly based on live observations and comparisons of their highly variable cell morphologies. It makes accurate identification and quantification of rumen Ciliates very difficult if not impossible. The development of fluorescence in situ hybridization (FISH), established as a technique to identify prokaryotes and eukaryotes using ribosomal RNA-targeted fluorescently labeled oligonucleotide probes, is a promising approach to identify and investigate rumen Ciliates. The present study, part of CIMES (Ciliates as monitors for environmental safety), a project sponsored by the European Commission, shows the problems of applying FISH on rumen Ciliates and how to solve them. Tests resulted in a new protocol, which recommends para-formaldehyde and formaldehyde at 1–2% final concentration to preserve the Ciliates before applying FISH. Furthermore, seven new oligonucleotide probes could be developed and successfully be tested to identify different rumen ciliate taxa of the order Entodiniomorphida (class Litostomatea) by applying FISH. It is also shown, how FISH together with confocal laser scanning microscopy can improve analyses of ciliate cell morphologies. Thanks to Peter Pristas, Peter Javorsky, Ralf Einspanier, Susanne Ulbrich (providing rumen samples), Seung Yeo Moon-van der Staay, Georg van der Staay (providing unpublished 18S-rDNA sequences), the European Commission (financial support, project QLK3-CT-2002-02151).
J. Fried - One of the best experts on this subject based on the ideXlab platform.
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identification of rumen Ciliates using small subunit ribosomal rna 18s rrna targeted oligonucleotide probes and fluorescence in situ hybridization fish
Journal of Eukaryotic Microbiology, 2005Co-Authors: J. Fried, Nadine A Thomas, Johannes H P Hackstein, Neil R. Mcewan, S. Kišidayová, Jamie C Newbold, Wolfgang Ludwig, Tadeusz Michalowski, S Weber, Karlheinz SchleiferAbstract:Up to half of the biomass in the rumen can be represented by Ciliates, which play an important role in the digestion processes of their hosts. In the literature, very little information can be found on determination of the diversity of complex rumen ciliate communities. The identification of these fast moving protists is mainly based on live observations and comparisons of their highly variable cell morphologies. It makes accurate identification and quantification of rumen Ciliates very difficult if not impossible. The development of fluorescence in situ hybridization (FISH), established as a technique to identify prokaryotes and eukaryotes using ribosomal RNA-targeted fluorescently labeled oligonucleotide probes, is a promising approach to identify and investigate rumen Ciliates. The present study, part of CIMES (Ciliates as monitors for environmental safety), a project sponsored by the European Commission, shows the problems of applying FISH on rumen Ciliates and how to solve them. Tests resulted in a new protocol, which recommends para-formaldehyde and formaldehyde at 1–2% final concentration to preserve the Ciliates before applying FISH. Furthermore, seven new oligonucleotide probes could be developed and successfully be tested to identify different rumen ciliate taxa of the order Entodiniomorphida (class Litostomatea) by applying FISH. It is also shown, how FISH together with confocal laser scanning microscopy can improve analyses of ciliate cell morphologies. Thanks to Peter Pristas, Peter Javorsky, Ralf Einspanier, Susanne Ulbrich (providing rumen samples), Seung Yeo Moon-van der Staay, Georg van der Staay (providing unpublished 18S-rDNA sequences), the European Commission (financial support, project QLK3-CT-2002-02151).
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Identification of rumen Ciliates using small subunit ribosomal RNA (18S‐rRNA)‐targeted oligonucleotide probes and fluorescence in situ hybridization (FISH)
Journal of Eukaryotic Microbiology, 2005Co-Authors: J. Fried, Nadine A Thomas, Johannes H P Hackstein, Tadeusz Michałowski, Neil R. Mcewan, S. Kišidayová, Jamie C Newbold, Wolfgang Ludwig, S Weber, Karlheinz SchleiferAbstract:Up to half of the biomass in the rumen can be represented by Ciliates, which play an important role in the digestion processes of their hosts. In the literature, very little information can be found on determination of the diversity of complex rumen ciliate communities. The identification of these fast moving protists is mainly based on live observations and comparisons of their highly variable cell morphologies. It makes accurate identification and quantification of rumen Ciliates very difficult if not impossible. The development of fluorescence in situ hybridization (FISH), established as a technique to identify prokaryotes and eukaryotes using ribosomal RNA-targeted fluorescently labeled oligonucleotide probes, is a promising approach to identify and investigate rumen Ciliates. The present study, part of CIMES (Ciliates as monitors for environmental safety), a project sponsored by the European Commission, shows the problems of applying FISH on rumen Ciliates and how to solve them. Tests resulted in a new protocol, which recommends para-formaldehyde and formaldehyde at 1–2% final concentration to preserve the Ciliates before applying FISH. Furthermore, seven new oligonucleotide probes could be developed and successfully be tested to identify different rumen ciliate taxa of the order Entodiniomorphida (class Litostomatea) by applying FISH. It is also shown, how FISH together with confocal laser scanning microscopy can improve analyses of ciliate cell morphologies. Thanks to Peter Pristas, Peter Javorsky, Ralf Einspanier, Susanne Ulbrich (providing rumen samples), Seung Yeo Moon-van der Staay, Georg van der Staay (providing unpublished 18S-rDNA sequences), the European Commission (financial support, project QLK3-CT-2002-02151).
Senjie Lin - One of the best experts on this subject based on the ideXlab platform.
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prevalent ciliate symbiosis on copepods high genetic diversity and wide distribution detected using small subunit ribosomal rna gene
PLOS ONE, 2012Co-Authors: Zhiling Guo, Sheng Liu, Yousong Huang, Guangxing Liu, Huan Zhang, Senjie LinAbstract:Toward understanding the genetic diversity and distribution of copepod-associated symbiotic Ciliates and the evolutionary relationships with their hosts in the marine environment, we developed a small subunit ribosomal RNA gene (18S rDNA)-based molecular method and investigated the genetic diversity and genotype distribution of the symbiotic Ciliates on copepods. Of the 10 copepod species representing six families collected from six locations of Pacific and Atlantic Oceans, 9 were found to harbor ciliate symbionts. Phylogenetic analysis of the 391 ciliate 18S rDNA sequences obtained revealed seven groups (ribogroups), six (containing 99% of all the sequences) belonging to subclass Apostomatida, the other clustered with peritrich ciliate Vorticella gracilis. Among the Apostomatida groups, Group III were essentially identical to Vampyrophrya pelagica, and the other five groups represented the undocumented Ciliates that were close to Vampyrophrya/Gymnodinioides/Hyalophysa. Group VI Ciliates were found in all copepod species but one (Calanus sinicus), and were most abundant among all ciliate sequences obtained, indicating that they are the dominant symbiotic Ciliates universally associated with copepods. In contrast, some ciliate sequences were found only in some of the copepods examined, suggesting the host selectivity and geographic differentiation of Ciliates, which requires further verification by more extensive sampling. Our results reveal the wide occurrence and high genetic diversity of symbiotic Ciliates on marine copepods and highlight the need to systematically investigate the host- and geography-based genetic differentiation and ecological roles of these Ciliates globally.
S Weber - One of the best experts on this subject based on the ideXlab platform.
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identification of rumen Ciliates using small subunit ribosomal rna 18s rrna targeted oligonucleotide probes and fluorescence in situ hybridization fish
Journal of Eukaryotic Microbiology, 2005Co-Authors: J. Fried, Nadine A Thomas, Johannes H P Hackstein, Neil R. Mcewan, S. Kišidayová, Jamie C Newbold, Wolfgang Ludwig, Tadeusz Michalowski, S Weber, Karlheinz SchleiferAbstract:Up to half of the biomass in the rumen can be represented by Ciliates, which play an important role in the digestion processes of their hosts. In the literature, very little information can be found on determination of the diversity of complex rumen ciliate communities. The identification of these fast moving protists is mainly based on live observations and comparisons of their highly variable cell morphologies. It makes accurate identification and quantification of rumen Ciliates very difficult if not impossible. The development of fluorescence in situ hybridization (FISH), established as a technique to identify prokaryotes and eukaryotes using ribosomal RNA-targeted fluorescently labeled oligonucleotide probes, is a promising approach to identify and investigate rumen Ciliates. The present study, part of CIMES (Ciliates as monitors for environmental safety), a project sponsored by the European Commission, shows the problems of applying FISH on rumen Ciliates and how to solve them. Tests resulted in a new protocol, which recommends para-formaldehyde and formaldehyde at 1–2% final concentration to preserve the Ciliates before applying FISH. Furthermore, seven new oligonucleotide probes could be developed and successfully be tested to identify different rumen ciliate taxa of the order Entodiniomorphida (class Litostomatea) by applying FISH. It is also shown, how FISH together with confocal laser scanning microscopy can improve analyses of ciliate cell morphologies. Thanks to Peter Pristas, Peter Javorsky, Ralf Einspanier, Susanne Ulbrich (providing rumen samples), Seung Yeo Moon-van der Staay, Georg van der Staay (providing unpublished 18S-rDNA sequences), the European Commission (financial support, project QLK3-CT-2002-02151).
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Identification of rumen Ciliates using small subunit ribosomal RNA (18S‐rRNA)‐targeted oligonucleotide probes and fluorescence in situ hybridization (FISH)
Journal of Eukaryotic Microbiology, 2005Co-Authors: J. Fried, Nadine A Thomas, Johannes H P Hackstein, Tadeusz Michałowski, Neil R. Mcewan, S. Kišidayová, Jamie C Newbold, Wolfgang Ludwig, S Weber, Karlheinz SchleiferAbstract:Up to half of the biomass in the rumen can be represented by Ciliates, which play an important role in the digestion processes of their hosts. In the literature, very little information can be found on determination of the diversity of complex rumen ciliate communities. The identification of these fast moving protists is mainly based on live observations and comparisons of their highly variable cell morphologies. It makes accurate identification and quantification of rumen Ciliates very difficult if not impossible. The development of fluorescence in situ hybridization (FISH), established as a technique to identify prokaryotes and eukaryotes using ribosomal RNA-targeted fluorescently labeled oligonucleotide probes, is a promising approach to identify and investigate rumen Ciliates. The present study, part of CIMES (Ciliates as monitors for environmental safety), a project sponsored by the European Commission, shows the problems of applying FISH on rumen Ciliates and how to solve them. Tests resulted in a new protocol, which recommends para-formaldehyde and formaldehyde at 1–2% final concentration to preserve the Ciliates before applying FISH. Furthermore, seven new oligonucleotide probes could be developed and successfully be tested to identify different rumen ciliate taxa of the order Entodiniomorphida (class Litostomatea) by applying FISH. It is also shown, how FISH together with confocal laser scanning microscopy can improve analyses of ciliate cell morphologies. Thanks to Peter Pristas, Peter Javorsky, Ralf Einspanier, Susanne Ulbrich (providing rumen samples), Seung Yeo Moon-van der Staay, Georg van der Staay (providing unpublished 18S-rDNA sequences), the European Commission (financial support, project QLK3-CT-2002-02151).