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1. Identification of Oxygen-Responsive Transcripts in the Silage Inoculant Lactobacillus buchneri CD034 by RNA Sequencing.

Author

Felix Gregor Eikmeyer, Stefan Heinl, Hans Marx, Alfred Pühler, Reingard Grabherr, and Andreas Schlüter

Subjects
Medicine, Science
Abstract

The Lactobacillus buchneri CD034 strain, known to improve the ensiling process of green fodder and the quality of the silage itself was transcriptionally analyzed by sequencing of transcriptomes isolated under anaerobic vs. aerobic conditions. L. buchneri CD034 was first cultivated under anaerobic conditions and then shifted to aerobic conditions by aeration with 21% oxygen. Cultivations already showed that oxygen was consumed by L. buchneri CD034 after aeration of the culture while growth of L. buchneri CD034 was still observed. RNA sequencing data revealed that irrespective of the oxygen status of the culture, the most abundantly transcribed genes are required for basic cell functions such as protein biosynthesis, energy metabolism and lactic acid fermentation. Under aerobic conditions, 283 genes were found to be transcriptionally up-regulated while 198 genes were found to be down-regulated (p-value < 0.01). Up-regulated genes i. a. play a role in oxygen consumption via oxidation of pyruvate or lactate (pox, lctO). Additionally, genes encoding proteins required for decomposition of reactive oxygen species (ROS) such as glutathione reductase or NADH peroxidase were also found to be up-regulated. Genes related to pH homeostasis and redox potential balance were found to be down-regulated under aerobic conditions. Overall, genes required for lactic acid fermentation were hardly affected by the growth conditions applied. Genes identified to be differentially transcribed depending on the aeration status of the culture are suggested to specify the favorable performance of the strain in silage formation.

Published
2015
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2. Sequence of Two Plasmids from Clostridium perfringens Chicken Necrotic Enteritis Isolates and Comparison with C. perfringens Conjugative Plasmids

Author

Felix Gregor Eikmeyer, Marcio C. Costa, Valeria R. Parreira, Jochen Blom, and John F. Prescott

Subjects
Bacterial Diseases, Clostridium perfringens, Applied Microbiology, lcsh:Medicine, medicine.disease_cause, law.invention, Plasmid, law, Gene Order, lcsh:Science, Polymerase chain reaction, Phylogeny, Genetics, Multidisciplinary, Ecology, Animal Models, Veterinary Bacteriology, Chicken, Enteritis, Infectious Diseases, Veterinary Diseases, Conjugation, Genetic, Recombinant DNA, Medicine, Research Article, Biotechnology, Plasmids, Sequence analysis, Molecular Sequence Data, Locus (genetics), Biology, Microbiology, Microbial Ecology, Model Organisms, medicine, Animals, Gene, lcsh:R, Sequence Analysis, DNA, Genes, Bacterial, Mutation, Multilocus sequence typing, lcsh:Q, Veterinary Science, Chickens, Multilocus Sequence Typing
Abstract

Twenty-six isolates of Clostridium perfringens of different MLST types from chickens with necrotic enteritis (NE) (15 netB-positive) or from healthy chickens (6 netB-positive, 5 netB-negative) were found to contain 1–4 large plasmids, with most netB-positive isolates containing 3 large and variably sized plasmids which were more numerous and larger than plasmids in netB-negative isolates. NetB and cpb2 were found on different plasmids consistent with previous studies. The pathogenicity locus NELoc1, which includes netB, was largely conserved in these plasmids whereas NeLoc3, present in the cpb2 containing plasmids, was less well conserved. A netB-positive and a cpb2-positive plasmid were likely to be conjugative, and the plasmids were completely sequenced. Both plasmids possessed the intact tcp conjugative region characteristic of C. perfringens conjugative plasmids. Comparative genomic analysis of nine CpCPs, including the two plasmids described here, showed extensive gene rearrangements including pathogenicity locus and accessory gene insertions around rather than within the backbone region. The pattern that emerges from this analysis is that the major toxin-containing regions of the variety of virulence-associated CpCPs are organized as complex pathogenicity loci. How these different but related CpCPs can co-exist in the same host has been an unanswered question. Analysis of the replication-partition region of these plasmids suggests that this region controls plasmid incompatibility, and that CpCPs can be grouped into at least four incompatibility groups.

Published
2012

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