Deficiencies in thiamine (vitamin B1) cause a host of neurological and reproductive impairments y... more Deficiencies in thiamine (vitamin B1) cause a host of neurological and reproductive impairments yielding morbidity and mortality across environmental and clinical realms. In a technique analogous to immunomagnetic separation, we introduce the use of thiamine periplasmic binding protein (TBP)-conjugated magnetic beads to isolate thiamine from complex matrices. TBP expressed in Escherichia coli is highly specific to thiamine and provides an alternative to antibodies for this non-immunogenic target. After incubation with the sample and removal of unbound matrix constituents, thiamine is simultaneously released and converted to its fluorescent oxidation product thiochrome by alkaline potassium ferricyanide. Subsequent measurement of fluorescence at thiochrome-specific wavelengths provides a second layer of specificity for the detection of thiamine. Thiamine could be quantified at concentrations as low as 5 nM ranging up to 240 nM. Within, we apply this technique to selectively capture and quantify thiamine in complex salmonid fish egg and tissue matrices. Our results showed no measurable non-specific binding to the beads by endogenous fluorophores in the fish egg matrix. Thiamine levels as low as 0.2 nmol/g of fish egg can be detected using this approach, which is sufficient to assess deficiencies causing morbidity and mortality in fish that occur at 1.0 nmol/g of egg. This practical method may find application in other resource limited settings for clinical, food, or dietary supplement analyses.
It is rare that there are molecular clues to the evolutionary origin of developmental traits. We ... more It is rare that there are molecular clues to the evolutionary origin of developmental traits. We have encountered an evolutionary juxtaposition that may explain the origin of the unique replicative morphology of Epulopiscium spp., the largest known bacteria, which reproduce by the internal production of multiple live offspring. We report here a 16S rRNA-based phylogenetic analysis of Metabacterium polyspora, a multiple-endospore-forming, uncultivated inhabitant of guinea pig cecum. Cells of M. polyspora were harvested from cecum contents by sedimentation in a Ficoll gradient and lysed. The bacterial 16S rRNA genes of this lysate were amplified by PCR. Sequence analysis of the cloned PCR products revealed two dominant, closely related 16S rRNA types. In situ hybridization of cecum contents with fluorescently labeled oligonucleotides, diagnostic of these two sequences, showed that they represent distinct strains of M. polyspora. Phylogenetic analyses of the sequences showed that M. po...
Gut microbiota has become an integral component of the host, and received increasing attention. H... more Gut microbiota has become an integral component of the host, and received increasing attention. However, for many domestic animals, information on the microbiota is insufficient and more effort should be exerted to manage the gastrointestinal bacterial community. Understanding the factors that influence the composition of microbial community in the host alimentary canal is essential to manage or improve the microbial community composition. In the present study, 16S rRNA gene sequence-based comparisons of the bacterial communities in the grass carp (Ctenopharyngodon idellus) intestinal contents and fish culture-associated environments are performed. The results show that the fish intestinal microbiota harbors many cellulose-decomposing bacteria, including sequences related to Anoxybacillus, Leuconostoc, Clostridium, Actinomyces, and Citrobacter. The most abundant bacterial operational taxonomic units (OTUs) in the grass carp intestinal content are those related to feed digestion. In ...
Cellulosilyticum lentocellum DSM 5427 is an anaerobic, endospore-forming member of the Firmicutes... more Cellulosilyticum lentocellum DSM 5427 is an anaerobic, endospore-forming member of the Firmicutes. We describe the complete genome sequence of this cellulose-degrading bacterium; originally isolated from estuarine sediment of a river that received both domestic and paper mill waste. Comparative genomics of cellulolytic clostridia will provide insight into factors that influence degradation rates.
Cellulosilyticum lentocellum DSM 5427 is an anaerobic, endospore-forming member of the Firmicutes... more Cellulosilyticum lentocellum DSM 5427 is an anaerobic, endospore-forming member of the Firmicutes . We describe the complete genome sequence of this cellulose-degrading bacterium, which was originally isolated from estuarine sediment of a river that received both domestic and paper mill waste. Comparative genomics of cellulolytic clostridia will provide insight into factors that influence degradation rates.
Epulopiscium sp. type B is an enormous intestinal symbiont of the surgeonfish Naso tonganus . Int... more Epulopiscium sp. type B is an enormous intestinal symbiont of the surgeonfish Naso tonganus . Intracellular offspring production in Epulopiscium shares features with endospore formation. Here, we characterize the spoIIE homolog in Epulopiscium . The timing of spoIIE gene expression and presence of interacting partners suggest that the activation of σ F occurs early in Epulopiscium offspring development.
Epulopiscium sp. type B, a member of the Firmicutes, is a large (up to 300 microm), cigar-shaped ... more Epulopiscium sp. type B, a member of the Firmicutes, is a large (up to 300 microm), cigar-shaped bacterial symbiont of surgeonfish that propagates itself by forming multiple intracellular offspring. This unusual form of reproduction is an apparent modification of a developmental program used by some Firmicutes to produce an endospore. At the onset of offspring formation, the Epulopiscium cell divides at both poles. The polar cells are engulfed by the larger mother cell and grow within the mother cell. At the final stages of development, the Epulopiscium mother cell lyses. Here we describe changes in Epulopiscium cell structure, focusing on mother cell DNA replication and cell death. DNA replication was examined by labeling cells with the nucleotide analog bromodeoxyuridine. As expected, DNA replication occurs in the developing offspring. However, well after passage of genetic information from parent to offspring is complete, DNA within the mother cell continues to replicate. Using fluorescence microscopy, we found that near the end of the offspring growth cycle, mother cell DNA disintegrates. The mother cell membrane and wall deteriorate as well. DNA replication within this terminally differentiated cell indicates the importance of mother cell nucleoids in cell maintenance and the development of offspring. The synchronized timing of mother cell deterioration within a population suggests that the Epulopiscium mother cell undergoes a programmed cell death. The programmed death of the mother cell may allow for the timely release of resources accumulated in the mother cell to provide nutrients to populations of these intestinal microbes and their host.
Deficiencies in thiamine (vitamin B1) cause a host of neurological and reproductive impairments y... more Deficiencies in thiamine (vitamin B1) cause a host of neurological and reproductive impairments yielding morbidity and mortality across environmental and clinical realms. In a technique analogous to immunomagnetic separation, we introduce the use of thiamine periplasmic binding protein (TBP)-conjugated magnetic beads to isolate thiamine from complex matrices. TBP expressed in Escherichia coli is highly specific to thiamine and provides an alternative to antibodies for this non-immunogenic target. After incubation with the sample and removal of unbound matrix constituents, thiamine is simultaneously released and converted to its fluorescent oxidation product thiochrome by alkaline potassium ferricyanide. Subsequent measurement of fluorescence at thiochrome-specific wavelengths provides a second layer of specificity for the detection of thiamine. Thiamine could be quantified at concentrations as low as 5 nM ranging up to 240 nM. Within, we apply this technique to selectively capture and quantify thiamine in complex salmonid fish egg and tissue matrices. Our results showed no measurable non-specific binding to the beads by endogenous fluorophores in the fish egg matrix. Thiamine levels as low as 0.2 nmol/g of fish egg can be detected using this approach, which is sufficient to assess deficiencies causing morbidity and mortality in fish that occur at 1.0 nmol/g of egg. This practical method may find application in other resource limited settings for clinical, food, or dietary supplement analyses.
It is rare that there are molecular clues to the evolutionary origin of developmental traits. We ... more It is rare that there are molecular clues to the evolutionary origin of developmental traits. We have encountered an evolutionary juxtaposition that may explain the origin of the unique replicative morphology of Epulopiscium spp., the largest known bacteria, which reproduce by the internal production of multiple live offspring. We report here a 16S rRNA-based phylogenetic analysis of Metabacterium polyspora, a multiple-endospore-forming, uncultivated inhabitant of guinea pig cecum. Cells of M. polyspora were harvested from cecum contents by sedimentation in a Ficoll gradient and lysed. The bacterial 16S rRNA genes of this lysate were amplified by PCR. Sequence analysis of the cloned PCR products revealed two dominant, closely related 16S rRNA types. In situ hybridization of cecum contents with fluorescently labeled oligonucleotides, diagnostic of these two sequences, showed that they represent distinct strains of M. polyspora. Phylogenetic analyses of the sequences showed that M. po...
Gut microbiota has become an integral component of the host, and received increasing attention. H... more Gut microbiota has become an integral component of the host, and received increasing attention. However, for many domestic animals, information on the microbiota is insufficient and more effort should be exerted to manage the gastrointestinal bacterial community. Understanding the factors that influence the composition of microbial community in the host alimentary canal is essential to manage or improve the microbial community composition. In the present study, 16S rRNA gene sequence-based comparisons of the bacterial communities in the grass carp (Ctenopharyngodon idellus) intestinal contents and fish culture-associated environments are performed. The results show that the fish intestinal microbiota harbors many cellulose-decomposing bacteria, including sequences related to Anoxybacillus, Leuconostoc, Clostridium, Actinomyces, and Citrobacter. The most abundant bacterial operational taxonomic units (OTUs) in the grass carp intestinal content are those related to feed digestion. In ...
Cellulosilyticum lentocellum DSM 5427 is an anaerobic, endospore-forming member of the Firmicutes... more Cellulosilyticum lentocellum DSM 5427 is an anaerobic, endospore-forming member of the Firmicutes. We describe the complete genome sequence of this cellulose-degrading bacterium; originally isolated from estuarine sediment of a river that received both domestic and paper mill waste. Comparative genomics of cellulolytic clostridia will provide insight into factors that influence degradation rates.
Cellulosilyticum lentocellum DSM 5427 is an anaerobic, endospore-forming member of the Firmicutes... more Cellulosilyticum lentocellum DSM 5427 is an anaerobic, endospore-forming member of the Firmicutes . We describe the complete genome sequence of this cellulose-degrading bacterium, which was originally isolated from estuarine sediment of a river that received both domestic and paper mill waste. Comparative genomics of cellulolytic clostridia will provide insight into factors that influence degradation rates.
Epulopiscium sp. type B is an enormous intestinal symbiont of the surgeonfish Naso tonganus . Int... more Epulopiscium sp. type B is an enormous intestinal symbiont of the surgeonfish Naso tonganus . Intracellular offspring production in Epulopiscium shares features with endospore formation. Here, we characterize the spoIIE homolog in Epulopiscium . The timing of spoIIE gene expression and presence of interacting partners suggest that the activation of σ F occurs early in Epulopiscium offspring development.
Epulopiscium sp. type B, a member of the Firmicutes, is a large (up to 300 microm), cigar-shaped ... more Epulopiscium sp. type B, a member of the Firmicutes, is a large (up to 300 microm), cigar-shaped bacterial symbiont of surgeonfish that propagates itself by forming multiple intracellular offspring. This unusual form of reproduction is an apparent modification of a developmental program used by some Firmicutes to produce an endospore. At the onset of offspring formation, the Epulopiscium cell divides at both poles. The polar cells are engulfed by the larger mother cell and grow within the mother cell. At the final stages of development, the Epulopiscium mother cell lyses. Here we describe changes in Epulopiscium cell structure, focusing on mother cell DNA replication and cell death. DNA replication was examined by labeling cells with the nucleotide analog bromodeoxyuridine. As expected, DNA replication occurs in the developing offspring. However, well after passage of genetic information from parent to offspring is complete, DNA within the mother cell continues to replicate. Using fluorescence microscopy, we found that near the end of the offspring growth cycle, mother cell DNA disintegrates. The mother cell membrane and wall deteriorate as well. DNA replication within this terminally differentiated cell indicates the importance of mother cell nucleoids in cell maintenance and the development of offspring. The synchronized timing of mother cell deterioration within a population suggests that the Epulopiscium mother cell undergoes a programmed cell death. The programmed death of the mother cell may allow for the timely release of resources accumulated in the mother cell to provide nutrients to populations of these intestinal microbes and their host.
Uploads
Papers by Esther Angert