| 1. |
- Alves, G., et al.
(författare)
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Identification of Antibiotic Resistance Proteins via MiCId's Augmented Workflow. A Mass Spectrometry-Based Proteomics Approach
- 2022
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Ingår i: Journal of the American Society for Mass Spectrometry. - : American Chemical Society (ACS). - 1044-0305 .- 1879-1123. ; 33:6, s. 917-931
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Tidskriftsartikel (refereegranskat)abstract
- Fast and accurate identifications of pathogenic bacteria along with their associated antibiotic resistance proteins are of paramount importance for patient treatments and public health. To meet this goal from the mass spectrometry aspect, we have augmented the previously published Microorganism Classification and Identification (MiCId) workflow for this capability. To evaluate the performance of this augmented workflow, we have used MS/MS datafiles from samples of 10 antibiotic resistance bacterial strains belonging to three different species: Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. The evaluation shows that MiCId's workflow has a sensitivity value around 85% (with a lower bound at about 72%) and a precision greater than 95% in identifying antibiotic resistance proteins. In addition to having high sensitivity and precision, MiCId's workflow is fast and portable, making it a valuable tool for rapid identifications of bacteria as well as detection of their antibiotic resistance proteins. It performs microorganismal identifications, protein identifications, sample biomass estimates, and antibiotic resistance protein identifications in 6-17 min per MS/MS sample using computing resources that are available in most desktop and laptop computers. We have also demonstrated other use of MiCId's workflow. Using MS/MS data sets from samples of two bacterial clonal isolates, one being antibiotic-sensitive while the other being multidrug-resistant, we applied MiCId's workflow to investigate possible mechanisms of antibiotic resistance in these pathogenic bacteria; the results showed that MiCId's conclusions agree with the published study.
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| 2. |
- Carvalheira, A., et al.
(författare)
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Acinetobacter portensis sp. Nov. and acinetobacter guerrae sp. nov., isolated from raw meat
- 2020
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Ingår i: International journal of systematic and evolutionary microbiology. - : Microbiology Society. - 1466-5026 .- 1466-5034. ; 70:8, s. 4544-4554
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Tidskriftsartikel (refereegranskat)abstract
- The taxonomic status of six strains of Acinetobacter obtained from meat samples, collected from supermarkets in Porto, Por-tugal, was investigated using polyphasic analysis. Partial rpoB sequence similarities lower than 95 % to other Acinetobacter species with validly published names led to the hypothesis that these strains represented novel species. This was confirmed based on comparative multilocus sequence analysis, which included the gyrB, recA and 16S rRNA genes, revealing that these strains represented two coherent lineages that were distinct from each other and from all known species. The names Acine-tobacter portensis sp. nov. (comprising four strains) and Acinetobacter guerrae sp. nov. (comprising two strains) are proposed for these novel species. The species status of these two groups was confirmed by low (below 95 %) whole-genome sequence average nucleotide identity values and low (below 70 %) digital DNA–DNA hybridization similarities between the whole-genome sequences of the proposed type strains of each novel species and the representatives of the known Acinetobacter species. Phylogenomic treeing from core genome analysis supported these results. The coherence of each new species lineage was supported by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry differentiation of the species at the protein level, by cellular fatty acid profiles, and by unique and differential combinations of metabolic and physiological properties shared by each novel species. The type strain of A. portensis sp. nov. is AC 877T (=CCUG 68672T=CCM 8789T) and the type strain of A. guerrae sp. nov. is AC 1271T (=CCUG 68674T=CCM 8791T). © 2020 The Authors.
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| 3. |
- Crespi, S., et al.
(författare)
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Legionella maioricensis sp. nov., a new species isolated from the hot water distribution systems of a hospital and a shopping center during routine sampling
- 2023
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Ingår i: International Journal of Systematic and Evolutionary Microbiology. - : Microbiology Society. - 1466-5026 .- 1466-5034. ; 73:1
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Tidskriftsartikel (refereegranskat)abstract
- Two Legionella- like strains isolated from hot water distribution systems in 2012 have been characterized phenotypically, bio-chemically and genomically in terms of DNA relatedness. Both strains, HCPI- 6T and EUR- 108, exhibited biochemical pheno-typic profiles typical of Legionella species. Cells were Gram-negative motile rods which grew on BCYE alpha agar but not on blood agar and displayed phenotypic characteristics typical of the family Legionellaceae, including a requirement for L-cysteine and testing catalase positive. Both strains were negative for oxidase, urease, nitrate reduction and hippurate negative, and non -fermentative. The major ubiquinone was Q12 (59.4 % HCPI- 6T) and the dominant fatty acids were C16:1 omega 7c (28.4 % HCPI- 6T, 216 % EUR- 108), C16: 0 iso (222.5 % and 213 %) and C15: 0 anteiso (19.5 % and 223.5 %, respectively). The percent G+C content of genomic DNA was determined to be 39.3 mol %. The 16S rRNA gene, mip sequence and comparative genome sequence -based analyses (average nucleotide identity, ANI; digital DNA-DNA hybridization, dDDH; and phylogenomic treeing) demonstrated that the strains represent a new species of the genus Legionella. The analysis based on the 16S rRNA gene sequences showed that the sequence similarities for both strains ranged from 98.8-90.1 % to other members of the genus. The core genome- based phylogenomic tree (protein-concatemer tree based on concatenation of 418 proteins present in single copy) revealed that these two strains clearly form a separate cluster within the genus Legionella. ANI and dDDH values confirmed the distinctiveness of the strains. Based on the genomic, genotypic and phenotypic findings from a polyphasic study, the isolates are considered to represent a single novel species, for which the name Legionella maioricensis sp. nov. is proposed. The type strain is HCPI- 6T (=CCUG 75071T=CECT 30569T).
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| 4. |
- Jaen-Luchoro, Daniel, et al.
(författare)
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Corynebacterium genitalium sp. nov., nom. rev. and Corynebacterium pseudogenitalium sp. nov., nom. rev., two old species of the genus Corynebacterium described from clinical and environmental samples
- 2023
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Ingår i: Research in Microbiology. - : Elsevier BV. - 0923-2508. ; 174:1-2
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Tidskriftsartikel (refereegranskat)abstract
- Two Corynebacterium species were proposed decades ago, isolated from clinical samples and divided into biovars: "Corynebacterium genitalium" biovars I-V and "Corynebacterium pseudogenitalium" biovars C1 -C6. Several biovars have been re-classified as new species. Nevertheless, biovar I and C5, together with their respective specific epithets "Corynebacterium genitalium" and "Corynebacterium pseudogenitalium", remained not validly published after more than 40 years. Several more strains, temptatively classified as "C. genitalium" biovar I and "Corynebacterium pseudogenitalium" C5, have been isolated from clinical and environmental samples. Both species presented Gram-positive, non-spore forming rod-shaped cells, able to grow aerobically with CO2. Core-genome analysis identified "C. genitalium" to be most closely related to Corynebacterium tuscaniense, Corynebacterium urinipleomorphum, Corynebacterium aquatimens and C appendicis, and Corynebacterium gottingense as the most closely related species to "C. pseudogenitalium". Comprehensive genomic, genotypic, phenotypic analyses, as well as chemotaxonomic, support the proposal for "C. genitalium" and "C. pseudogenitalium" as distinct species within the genus Corynebacterium. The designated type strains of the two species are Furness 392-1T = ATCC 33030T = CCUG 38989T = CCM 9178T = DSM 113155T for C. genitalium sp. nov., nom. rev., and Furness 162-C2T = ATCC 33039T = CCUG 27540T = CCM 9177T = DSM 113154T for C. pseudogenitalium sp. nov., nom. rev. (c) 2022 The Author(s). Published by Elsevier Masson SAS on behalf of Institut Pasteur. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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| 5. |
- Jaen-Luchoro, Daniel, et al.
(författare)
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Knockout of Targeted Plasmid-Borne fl-Lactamase Genes in an Extended-Spectrum-fl-Lactamase-Producing Escherichia coli Strain: Impact on Resistance and Proteomic Profile
- 2023
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Ingår i: Microbiology Spectrum. - : American Society for Microbiology. - 2165-0497. ; 11:1
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Tidskriftsartikel (refereegranskat)abstract
- Resistance to beta-lactams is known to be multifactorial, although the underlying mechanisms are not well established. The aim of our study was to develop a system for assessing the phenotypic and proteomic responses of bacteria to antibiotic stress as a result of the loss of selected antimicrobial resistance genes. We applied homologous recombination to knock out plasmid-borne beta-lactamase genes (bla(OXA-1), bla(TEM-1), and bla(CTX-M15)) in Escherichia coli CCUG 73778, generating knockout clone variants lacking the respective deleted beta-lactamases. Quantitative proteomic analyses were performed on the knockout variants and the wild-type strain, using bottom-up liquid chromatography tandem mass spectrometry (LC-MS/MS), after exposure to different concentrations of cefadroxil. Loss of the bla(CTX-M-15) gene had the greatest impact on the resulting protein expression dynamics, while losses of bla(OXA-1) and bla(TEM-1) affected fewer proteins' expression levels. Proteins involved in antibiotic resistance, cell membrane integrity, stress, and gene expression and unknown function proteins exhibited differential expression. The present study provides a framework for studying protein expression in response to antibiotic exposure and identifying the genomic, proteomic, and phenotypic impacts of resistance gene loss.
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| 6. |
- Méndez, V., et al.
(författare)
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Complete genome sequence of the hydrocarbon-degrading strain achromobacter sp. B7, isolated during petroleum hydrocarbon bioremediation in the valparaiso region, Chile
- 2018
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Ingår i: Microbiology Resource Announcements. - 2576-098X. ; 7:19
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Tidskriftsartikel (refereegranskat)abstract
- Achromobacter sp. strain B7 (= CCUG 72081) was isolated from a diesel-polluted soil from the Valparaiso Region, Chile, subjected to bioremediation with a hydrocarbon-degrading enrichment. The complete genome sequence of Achromobacter sp. B7 has been determined to have a size of 6.24 Mb, 5,578 coding sequences, 57 tRNAs, and a G+C content of 64.8%. © 2018 Méndez et al.
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| 7. |
- Pain, M., et al.
(författare)
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Staphylococcus borealis sp. nov., isolated from human skin and blood
- 2020
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Ingår i: International journal of systematic and evolutionary microbiology. - : Microbiology Society. - 1466-5026 .- 1466-5034. ; 70:12, s. 6067-6078
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Tidskriftsartikel (refereegranskat)abstract
- When analysing a large cohort of Staphylococcus haemolyticus, using whole-genome sequencing, five human isolates (four from the skin and one from a blood culture) with aberrant phenotypic and genotypic traits were identified. They were phenotypically similar with yellow colonies, nearly identical 16S rRNA gene sequences and initially speciated as S. haemolyticus based on 16S rRNA gene sequence and MALDI-TOF MS. However, compared to S. haemolyticus, these five strains demonstrate: (i) considerable phylogenetic distance with an average nucleotide identity <95 % and inferred DNA-DNA hybridization <70%; (ii) a pigmented phenotype; (iii) urease production; and (iv) different fatty acid composition. Based on the phenotypic and genotypic results, we conclude that these strains represent a novel species, for which the name Staphylococcus borealis sp. nov. is proposed. The novel species belong to the genus Staphylococcus and is coagulase- and oxidase-negative and catalase-positive. The type strain, 51-48(T), is deposited in the Culture Collection University of Gothenburg (CCUG 73747(T)) and in the Spanish Type Culture Collection (CECT 30011(T)).
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| 8. |
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