TL;DR: The taxonomic status of 14 strains of members of the genus Acinetobacter isolated from floral nectar of wild Mediterranean insect-pollinated plants, which did not belong to any previously described species within this genus, was investigated following a polyphasic approach and confirmed that these strains formed two separate lineages.
Abstract: The taxonomic status of 14 strains of members of the genus Acinetobacter isolated from floral nectar of wild Mediterranean insect-pollinated plants, which did not belong to any previously described species within this genus, was investigated following a polyphasic approach. Confirmation that these strains formed two separate lineages within the genus Acinetobacter was obtained from comparative analysis of the partial sequences of the 16S rRNA gene and the gene encoding the β-subunit of RNA polymerase (rpoB), DNA-DNA reassociation data, determination of the DNA G+C content and physiological tests. The names Acinetobacter nectaris sp. nov. and Acinetobacter boissieri sp. nov. are proposed. The type strain of A. nectaris sp. nov. is SAP 763.2(T) ( = LMG 26958(T) = CECT 8127(T)) and that of A. boissieri sp. nov. is SAP 284.1(T) ( = LMG 26959(T) = CECT 8128(T)).
TL;DR: Phenotypic, phylogenetic and chemotaxonomic data, including low DNA–DNA relatedness with closely related type strains, supported that strain WB1T represents a distinct novel species in the genus Acinetobacter.
Abstract: A Gram-negative bacterial strain, designated WB1(T), was isolated from a domestic refrigerator in Guangzhou, PR China. Cells of strain WB1(T) were oxidase-negative, catalase-positive, strictly aerobic, non-spore-forming and non-motile coccobacilli with peritrichous fimbriae-like structures. The strain was able to grow at 10-40 °C with optimum growth at 28-30 °C, pH 6.0-8.0 (optimum, pH 7.0) and 0-6 % NaCl (w/v, optimum, 0.5 %). Phylogenetic analyses based on 16S rRNA gene and rpoB gene sequences revealed that strain WB1(T) belonged to the genus Acinetobacter and was most closely related to A. indicus DSM 25388(T) (97.2 % 16S rRNA gene sequence similarity) and A. radioresistens NBRC 102413(T) (96.8 %). The DNA G + C content of strain WB1(T) was 46.74 ± 0.04 mol % and the major fatty acids comprised summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), C18:1 ω9c, C16:0 and C12:0. The predominant respiratory quinone was identified as Q-9 and the polar lipids as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and an unidentified phospholipid. Phenotypic, phylogenetic and chemotaxonomic data, including low DNA-DNA relatedness with closely related type strains, supported that strain WB1(T) represents a distinct novel species in the genus Acinetobacter, for which the name Acinetobacter refrigeratorensis sp. nov. was proposed. The type strain is WB1(T) (=GIMCC 1.663(T) = CCTCC AB 2014197(T) = KCTC 42011(T)).
6 citations
Cites background from "Acinetobacter nectaris sp. nov. and..."
...specimens [21, 23], floral nectar and canker bark of plants [1, 16, 17], raw wastewater [30] and soil [5]....
TL;DR: The taxonomic position and structure of a novel, taxonomically unique group of 26 Acinetobacter strains, provisionally designated Taxon 24 (T24), were defined in this article.
TL;DR: The author states that the aim of this book is to provide a history of statistical analysis of statistical methods and their applications in the field of quantitative anthropology.
Abstract: ................................................................................................................................ ii Acknowledgments ................................................................................................................ iv Abbreviations ....................................................................................................................... vi Table of content .................................................................................................................. viii List of figures ....................................................................................................................... xii List of tables ........................................................................................................................ xiv Chapter
TL;DR: The results highlight that collection time, collection location, and mosquito species each affect aspects of mosquito microbial diversity, but their importance is context dependent, and demonstrate that these variables have differing impacts on mosquito diversity and mosquito microbial Diversity.
Abstract: The mosquito microbiota affects many aspects of mosquito biology including development and reproduction. It also strongly impacts interactions between the mosquito host and pathogens that cause important disease in humans, such as dengue and malaria. Critically, the mosquito microbiota is highly diverse and can vary in composition in response to multiple environmental variables, but these effects are not always consistent. Understanding how the environment shapes mosquito microbial diversity is a critical step in elucidating the ubiquity of key host-microbe-pathogen interactions in nature. To that end, we examined the role of time of collection, collection location and host species on mosquito microbial diversity by repeating collections at two-month intervals on a trapping grid spanning three distinct biomes. We then used 16S rRNA sequencing to compare the microbiomes of Aedes taeniorhynchus, Anopheles crucians, and Culex nigripalpus mosquitoes from those collections. We saw that mosquito diversity was strongly affected by both time and collection location. We also observed that microbial richness and diversity increased from March to May, and that An. crucians and Cx. nigripalpus had greater microbial diversity than Ae. taeniorhynchus. However, we also observed that collection location had no impact on microbial diversity except for significantly lower bacterial richness observed in mosquitoes collected from the mangrove wetlands. Our results highlight that collection time, collection location, and mosquito species each affect aspects of mosquito microbial diversity, but their importance is context dependent. We also demonstrate that these variables have differing impacts on mosquito diversity and mosquito microbial diversity. Our findings suggest that the environment likely plays an important but variable role in influencing the composition of the mosquito microbiota.
TL;DR: It is suggested that inhibition of AdeABC could prevent biofilm formation or colonisation in patients by A. baumannii and so provides a good target for drug discovery.
Abstract: Acinetobacter baumannii is a nosocomial pathogen and causes infections in hospitals worldwide. This organism is often multi-drug resistant (MDR), can persist in the environment and forms a biofilm on environmental surfaces and wounds.
This thesis describes research that investigates the role of the two component system AdeRS, which regulates production of the AdeABC MDR efflux pump. Its role in MDR, biofilm formation and virulence of A. baumannii was determined in mutants constructed for this study. Deletion of AdeRS or AdeABC resulted in increased susceptibility to antibiotics, decreased biofilm formation on biotic and abiotic surfaces and decreased virulence in a strain dependent manner. RNA-Seq revealed that loss of AdeRS or AdeB significantly altered the transcriptome, resulting in changed expression of many genes, notably those associated with antimicrobial resistance and virulence interactions.
Thjs study demonstrated the scope of AdeRS mediated regulation and suggests that inhibition of AdeABC could prevent biofilm formation or colonisation in patients by A. baumannii and so provides a good target for drug discovery. This study also highlighted the differences between A. baumannii strains and shows that conclusions for the species should not be drawn from the study of single strains.
TL;DR: The newest addition in MEGA5 is a collection of maximum likelihood (ML) analyses for inferring evolutionary trees, selecting best-fit substitution models, inferring ancestral states and sequences, and estimating evolutionary rates site-by-site.
Abstract: Comparative analysis of molecular sequence data is essential for reconstructing the evolutionary histories of species and inferring the nature and extent of selective forces shaping the evolution of genes and species. Here, we announce the release of Molecular Evolutionary Genetics Analysis version 5 (MEGA5), which is a user-friendly software for mining online databases, building sequence alignments and phylogenetic trees, and using methods of evolutionary bioinformatics in basic biology, biomedicine, and evolution. The newest addition in MEGA5 is a collection of maximum likelihood (ML) analyses for inferring evolutionary trees, selecting best-fit substitution models (nucleotide or amino acid), inferring ancestral states and sequences (along with probabilities), and estimating evolutionary rates site-by-site. In computer simulation analyses, ML tree inference algorithms in MEGA5 compared favorably with other software packages in terms of computational efficiency and the accuracy of the estimates of phylogenetic trees, substitution parameters, and rate variation among sites. The MEGA user interface has now been enhanced to be activity driven to make it easier for the use of both beginners and experienced scientists. This version of MEGA is intended for the Windows platform, and it has been configured for effective use on Mac OS X and Linux desktops. It is available free of charge from http://www.megasoftware.net.
39,110 citations
"Acinetobacter nectaris sp. nov. and..." refers methods in this paper
...Following these procedures, 1320 nt positions (98 % of the original alignment) remained for subsequent phylogenetic analysis using the neighbour-joining (NJ) method as implemented in the MEGA 5 software package (Tamura et al., 2011)....
TL;DR: A computerized method is presented that reduces to a certain extent the necessity of manually editing multiple alignments, makes the automation of phylogenetic analysis of large data sets feasible, and facilitates the reproduction of the final alignment by other researchers.
Abstract: The use of some multiple-sequence alignments in phylogenetic analysis, particularly those that are not very well conserved, requires the elimination of poorly aligned positions and divergent regions, since they may not be homologous or may have been saturated by multiple substitutions. A computerized method that eliminates such positions and at the same time tries to minimize the loss of informative sites is presented here. The method is based on the selection of blocks of positions that fulfill a simple set of requirements with respect to the number of contiguous conserved positions, lack of gaps, and high conservation of flanking positions, making the final alignment more suitable for phylogenetic analysis. To illustrate the efficiency of this method, alignments of 10 mitochondrial proteins from several completely sequenced mitochondrial genomes belonging to diverse eukaryotes were used as examples. The percentages of removed positions were higher in the most divergent alignments. After removing divergent segments, the amino acid composition of the different sequences was more uniform, and pairwise distances became much smaller. Phylogenetic trees show that topologies can be different after removing conserved blocks, particularly when there are several poorly resolved nodes. Strong support was found for the grouping of animals and fungi but not for the position of more basal eukaryotes. The use of a computerized method such as the one presented here reduces to a certain extent the necessity of manually editing multiple alignments, makes the automation of phylogenetic analysis of large data sets feasible, and facilitates the reproduction of the final alignment by other researchers.
8,757 citations
"Acinetobacter nectaris sp. nov. and..." refers methods in this paper
...0 (Hall, 1999) to ensure that all sequences had the same start and end point, and analysed with Gblocks (Castresana, 2000) to eliminate ambiguously aligned regions, using ‘allow gap positions5with half’, ‘minimum length of a block559 and default settings for all other options....
[...]
...…alignment was trimmed with BioEdit v. 7.0.9.0 (Hall, 1999) to ensure that all sequences had the same start and end point, and analysed with Gblocks (Castresana, 2000) to eliminate ambiguously aligned regions, using ‘allow gap positions5with half’, ‘minimum length of a block559 and default settings…...
[...]
...As with the 16S rRNA gene sequence, concatenated sequences of rpoB zones 1 and 2 of the nectar strains and type strains of other species of the genus Acinetobacter were included in a multiple alignment and analysed with Gblocks, which resulted in the selection of 843 nucleotide positions (99 % of the original alignment)....
[...]
...The resulting alignment was trimmed with BioEdit v. 7.0.9.0 (Hall, 1999) to ensure that all sequences had the same start and end point, and analysed with Gblocks (Castresana, 2000) to eliminate ambiguously aligned regions, using ‘allow gap positions5with half’, ‘minimum length of a block559 and default settings for all other options....
TL;DR: The Clustal series of programs, widely used in molecular biology for the multiple alignment of both nucleic acid and protein sequences and for preparing phylogenetic trees, are extended.
Abstract: The Clustal series of programs are widely used in molecular biology for the multiple alignment of both nucleic acid and protein sequences and for preparing phylogenetic trees. The popularity of the programs depends on a number of factors, including not only the accuracy of the results, but also the robustness, portability and user-friendliness of the programs. New features include NEXUS and FASTA format output, printing range numbers and faster tree calculation. Although, Clustal was originally developed to run on a local computer, numerous Web servers have been set up, notably at the EBI (European Bioinformatics Institute) (http://www.ebi.
5,300 citations
"Acinetobacter nectaris sp. nov. and..." refers methods in this paper
...The 16S rRNA gene sequences of the novel nectar strains and reference strains of members of the genus Acinetobacter and the family Moraxellaceae were included in a multiple alignment generated by CLUSTAL W (Chenna et al., 2003)....
TL;DR: High-performance liquid chromatography is a promising alternative for determining the G+C content of bacterial deoxyribonucleic acid (DNA) and may also be more accurate than indirect methods, such as the buoyant density and thermal denaturation methods.
Abstract: High-performance liquid chromatography is a promising alternative for determining the G+C content of bacterial deoxyribonucleic acid (DNA). The method which we evaluated involves enzymatic degradation of the DNA to nucleosides by P1 nuclease and bovine intestinal mucosa alkaline phosphatase, separation of the nucleosides by high-performance liquid chromatography, and calculation of the G+C content from the apparent ratios of deoxyguanosine and thymidine. Because the nucleosides are released from the DNA at different rates, incomplete degradation produces large errors in the apparent G+C content. For partially purified DNA, salts are a major source of interference in degradation. However, when the salts are carefully removed, the preparation and degradation of DNA contribute little error to the determination of G+C content. This method also requires careful selection of the chromatographic conditions to ensure separation of the major nucleosides from the nucleosides of modified bases and precise control of the flow rates. Both of these conditions are achievable with standard equipment and C18 reversed-phase columns. Then the method is precise, and the relative standard deviations of replicate measurements are close to 0.1%. It is also rapid, and a single measurement requires about 15 min. It requires small amounts of sample, and the G+C content can be determined from DNA isolated from a single bacterial colony. It is not affected by contamination with ribonucleic acid. Because this method yields a direct measurement, it may also be more accurate than indirect methods, such as the buoyant density and thermal denaturation methods. In addition, for highly purified DNA, the extent of modification can be determined.
4,685 citations
"Acinetobacter nectaris sp. nov. and..." refers methods in this paper
...The DNA was enzymically degraded into nucleosides and the nucleotidic base composition was determined by HPLC, according to the method of Mesbah et al. (1989)....