Artículos (Microbiología)
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Artículo ginmappeR: an unified approach for integrating gene and protein identifiers across biological sequence databases(Oxford Univ Press, 2024-08-29) Sola Espinosa, Fernando Luis; Ayala Hernández, Daniel; Pulido, Marina R.; Ayala, Rafael; López Cerero, Lorena; Hernández Salmerón, Inmaculada Concepción; Ruiz Cortés, David; Universidad de Sevilla. Departamento de Lenguajes y Sistemas Informáticos; Universidad de Sevilla. Departamento de Microbiología; Ministerio de Ciencia e Innovación, España; Instituto de Carlos III; Ministerio de UniversidadesThe proliferation of biological sequence data, due to developments in molecular biology techniques, has led to the creation of numerous open access databases on gene and protein sequencing. However, the lack of direct equivalence between identifiers across these databases difficults data integration. To address this challenge, we introduce ginmappeR, an integrated R package facilitating the translation of gene and protein identifiers between databases. By providing a unified interface, ginmappeR streamlines the integration of diverse data sources into biological workflows, so it enhances efficiency and user experience.Artículo Association of Microbiological Factors with Mortality in Escherichia Coli Bacteraemia Presenting with Sepsis/septic Shock: A Prospective Cohort Study(Elsevier, 2024) Maldonado, Natalia; López Hernández, Inmaculada; López-Cortes, Luis Eduardo; Martínez Pérez-Crespo, Pedro María; Retamar Gentil, Pilar; García Montaner, Andrea; De la Rosa Riestra, Sandra; Sousa Domínguez, Adrián; Goikoetxea, Josune; Pulido Navazo, Ángeles; Pascual Hernández, Álvaro; Rodríguez-Baño, Jesús; Universidad de Sevilla. Departamento de MicrobiologíaObjectives: This study aimed to determine the association of Escherichia coli microbiological factors with 30-day mortality in patients with bloodstream infection (BSI) presenting with a dysregulated response to infection (i.e. sepsis or septic shock). Methods: Whole-genome sequencing was performed on 224 E coli isolates of patients with sepsis/septic shock, from 22 Spanish hospitals. Phylogroup, sequence type, virulence, antibiotic resistance, and pathogenicity islands were assessed. A multivariable model for 30-day mortality including clinical and epidemiological variables was built, to which microbiological variables were hierarchically added. The predictive capacity of the models was estimated by the area under the receiver operating characteristic curve (AUROC) with 95% confidence intervals (CI). Results: Mortality at day 30 was 31% (69 patients). The clinical model for mortality included (adjusted OR; 95% CI) age (1.04; 1.02e1.07), Charlson index 3 (1.78; 0.95e3.32), urinary BSI source (0.30; 0.16 e0.57), and active empirical treatment (0.36; 0.11e1.14) with an AUROC of 0.73 (95% CI, 0.67e0.80). Addition of microbiological factors selected clone ST95 (3.64; 0.94e14.04), eilA gene (2.62; 1.14e6.02), and astA gene (2.39; 0.87e6.59) as associated with mortality, with an AUROC of 0.76 (0.69e0.82). Discussion: Despite having a modest overall contribution, some microbiological factors were associated with increased odds of death and deserve to be studied as potential therapeutic or preventive targets.Artículo Antimicrobial Activity of Anionic Bis(N-Heterocyclic Carbene) Silver Complexes(Multidisciplinary Digital Publishing Institute, 2024-09-27) Carrasco Carrasco, Carlos Jesús; Montilla Ramos, Francisco Javier; Villalobo Polo, Eduardo; Angulo, Manuel; Álvarez, Eleuterio; Galindo del Pozo, Agustín; Universidad de Sevilla. Departamento de Microbiología; Ministerio de Ciencia e Innovación (MICIN). España; Universidad de SevillaThe antimicrobial properties of a series of anionic bis(carbene) silver complexes Na3[Ag(NHCR)2] were investigated (2a–2g and 2c′, where NHCR is a 2,2′-(imidazol-2-ylidene)dicarboxylate-type N-heterocyclic carbene). The complexes were synthesized by the interaction of imidazolium dicarboxylate compounds with silver oxide in the presence of aqueous sodium hydroxide. Complexes 2f,g were characterized analytically and spectroscopically, and the ligand precursor 1f and complexes 2c and 2g were structurally identified by X-ray diffraction methods. The anions of 2c and 2g, [Ag(NHCR)2]3−, showed a typical linear disposition of Ccarbene-Ag-Ccarbene atoms and an uncommonly eclipsed conformation of carbene ligands. The antimicrobial properties of complexes 2a–g, which contains chiral (2b–2e and 2c′) and non-chiral derivatives (2a,f,g), were evaluated against Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, and a Gram-positive bacterium, Staphylococcus aureus. From the observed values of the minimal inhibitory concentration and minimal bactericidal concentration, complexes 2a and 2b showed the best antimicrobial activity against all strains. An interesting chirality–antimicrobial relationship was found, and eutomer 2c′ showed better activity than its enantiomer 2c against the three bacteria. Furthermore, these complexes were investigated experimentally and theoretically by 109Ag nuclear magnetic resonance, and the electronic and steric characteristics of the dianionic carbene ligands were also examined.Artículo Genetic Features of BEL-1-Producing and KPC-2-Producing E. coli from Hospital Wastewater: Human Source or Sewages Adaptation(Springer Nature, 2024) Romero Oraá, Laura; Pulido, Marina R.; Galán, Fatima; Palacios, María Victoria García; Pascual Hernández, Álvaro; López Cerero, Lorena; Universidad de Sevilla. Departamento de Microbiología; Ministerio de Economía y Competitividad (MINECO). EspañaHospital sewage is an ecosystem that facilitates the transfer of antibiotic and heavy metal resistance genes and the interaction of human and environmental bacteria. In this environment, we have detected the presence of 7 KPC-2 and BEL-1 co-producing E. coli isolates of two different clones over a 10-month period in the same hospital. All isolates carried blaKPC-2 and the operon mer on the same IncP plasmid of similar size and an IncN plasmid of different size each clone carrying blaBEL-1. Both IncN-blaBEL-1 plasmids shared a 77 kb region containing blaBEL-1 alongside with fosE, blaOXA-10 and aac(6’)-1b genes in a class 3 integron within a Tn3 transposon. The major IncN plasmid contained in addition a region homolog to P1-like bacteriophage RCS47, including the lytic RepL and lysogenic proteins, but other phage regions were incomplete. The characters such as the temporal persistence in sewage, the absence of colonized patients in the hospital or in the region, the presence of a p1 phage-plasmid fusion and the infrequent class 3 integron as genetic platform would indicate that BEL-1-producing isolates could have been generated in situ by adaptation to human sewage. Part of the microbiota in these discharges could be explained by the interactions of sewage ecosystems and not derive directly from the hospital.Artículo Whole-genome characterisation of Escherichia coli isolates from patients with bacteraemia presenting with sepsis or septic shock in Spain: a multicentre cross-sectional study(Elsevier, 2024-04) Maldonado, Natalia; López Hernández, Inmaculada; García Montaner, Andrea; López-Cortes, Luis Eduardo; Martínez Pérez-Crespo, Pedro María; Retamar Gentil, Pilar; Rodríguez-Baño, Jesús; Pascual Hernández, Álvaro; Universidad de Sevilla. Departamento de Medicina; Universidad de Sevilla. Departamento de MicrobiologíaBackground Escherichia coli is the most frequent cause of bloodstream infections (BSIs). About one-third of patients with BSIs due to E coli develop sepsis or shock. The objective of this study is to characterise the microbiological features of E coli blood isolates causing sepsis or septic shock to provide exploratory information for future diagnostic, preventive, or therapeutic interventions. Methods Ecolibloodisolatesfromamulticentrecross-sectionalstudyofpatientsolderthan14yearspresentingwithsepsis or septic shock (according to the Third International Consensus Definitions for Sepsis and Septic Shock criteria) from hospitals in Spain between Oct 4, 2016, and Oct 15, 2017, were studied by whole-genome sequencing. Phylogroups, sequence types (STs), serotype, FimH types, antimicrobial resistance (AMR) genes, pathogenicity islands, and virulence factors were identified. Susceptibility testing was performed by broth microdilution. The main outcome of this study was the characterisation of the E coli blood isolates in terms of population structure by phylogroups, groups (group 1: phylogroups B2, F, and G; group 2: A, B1, and C; group 3: D), and STs and distribution by geographical location and bloodstream infection source. Other outcomes were virulence score and prevalence of virulence-associated genes, pathogenicity islands, AMR, and AMR-associated genes. Frequencies were compared using χ2 or Fisher’s exact tests, and continuous variables using the Mann-Whitney test, with Bonferroni correction for multiple comparisons. Findings We analysed 224 isolates: 140 isolates (63%) were included in phylogenetic group 1, 52 (23%) in group 2, and 32 (14%) in group 3. 85 STs were identified, with four comprising 44% (n=98) of the isolates: ST131 (38 [17%]), ST73 (25 [11%]), ST69 (23 [10%]), and ST95 (12 [5%]). No significant differences in phylogroup or ST distribution were found according to geographical areas or source of bloodstream infection, except for ST95, which was more frequent in urinary tract infections than in other sources (11 [9%] of 116 vs 1 [1%] of 108, p=0⋅0045). Median virulence score was higher in group 1 (median 25⋅0 [IQR 20⋅5–29⋅0) than in group 2 (median 14⋅5[9⋅0–20⋅0]; p<0⋅0001) and group 3 (median 21 [16⋅5–23⋅0]; p<0⋅0001); prevalence of several pathogenicity islands was higher in group 1. No significant differences were found between phylogenetic groups in proportions of resistance to antibiotics. ST73 had higher median virulence score (32 [IQR 29–35]) than the other predominant clones (median range 21–28). Some virulence genes and pathogenicity islands were significantly associated with each ST. ST131 isolates had higher prevalence of AMR and a higher proportion of AMR genes, notably blaCTX-M-15 and blaOXA-1. Interpretation In this exploratory study, the population structure of E coli causing sepsis or shock was similar to previous studies that included all bacteraemic isolates. Virulence genes, pathogenicity islands, and AMR genes were not randomly distributed among phylogroups or STs. These results provide a comprehensive characterisation of invasive E coli isolates causing severe response syndrome. Future studies are required to determine the contribution of these microbiological factors to severe clinical presentation and worse outcomes in patients with E coli bloodstream infection.Artículo Implications of Two-component Systems EnvZ/OmpR and BaeS/BaeR in in Vitro Temocillin Resistance in Escherichia Coli(Oxford University Press, 2024) Pérez Palacios, Patricia; Rodríguez Ochoa, José Luis; Velázquez Escudero, Ana; Rodríguez-Baño, Jesús; Rodríguez Martínez, José Manuel; Pascual Hernández, Álvaro; Docobo Pérez, Fernando; Universidad de Sevilla. Departamento de Microbiología; Universidad de Sevilla. Departamento de Medicina; Junta de Andalucía; Sociedad Europea de Microbiología Clínica y Enfermedades Infecciosas (ECCMID)Background: BaeS/BaeR is a two-component system of Escherichia coli that controls the expression of porins and efflux pumps. Its role in beta-lactam resistance is limited. Objectives: To study the role of baeS/baeR two-component system in temocillin resistance in E. coli. Methods: E. coli strain BW25113 and single-gene deletion mutants related to two-component systems were collected from the KEIO collection. Double-gen deletion mutants were generated. Temocillin-resistant mutant frequencies were determined at 32 mg/L. E. coli BW25113 mutants were selected by selective pressure from serial passages. Biological costs were analysed by growth curves. Genomes of the generated mutants were sequenced. The expression level of the mdtA, mdtB, mdtC, acrD and tolC in the ΔbaeS mutant was determined by RT–PCR (with/without temocillin exposure). Results: The frequency of temocillin mutants ranged from 2.12 × 10−8 to 4.51 × 10−8 in single-porin mutants. No mutants were recovered from E. coli BW25113 (>10−9). Selection of temocillin-resistant variants by serial passage yielded mutants up to 128 mg/L. Mutations were found in the baeS gene. Temocillin MICs ranged from 4 to 32 mg/L (highest MICs for ΔbaeS and ΔompR). The efflux pumps mdtA, mdtB, mdtC and acrD pumps were overexpressed 3–10-fold in the presence of temocillin in ΔbaeS compared to control. Conclusions: Mutations in the sensor histidine kinase, baeS, may be involved in temocillin resistance through the expression of the efflux pumps mdtABC and acrD. In addition, the low mutation rate may be a good predictor of temocillin activity.Artículo Impact of Suppression of the SOS Response on Protein Expression in Clinical Isolates of Escherichia Coli under Antimicrobial Pressure of Ciprofloxacin(Frontiers Media S.A., 2024) Recacha, Esther; Kuropka, Benno; Díaz Díaz, Sara; García Montaner, Andrea; González Tortuero, Enrique; Docobo Pérez, Fernando; Rodríguez Rojas, Alexandro; Rodríguez Martínez, José Manuel; Universidad de Sevilla. Departamento de Microbiología; Instituto de Salud Carlos IIIIntroduction/objective: Suppression of the SOS response in combination with drugs damaging DNA has been proposed as a potential target to tackle antimicrobial resistance. The SOS response is the pathway used to repair bacterial DNA damage induced by antimicrobials such as quinolones. The extent of lexA-regulated protein expression and other associated systems under pressure of agents that damage bacterial DNA in clinical isolates remains unclear. The aim of this study was to assess the impact of this strategy consisting on suppression of the SOS response in combination with quinolones on the proteome profile of Escherichia coli clinical strains. Materials and methods: Five clinical isolates of E. coli carrying different chromosomally- and/or plasmid-mediated quinolone resistance mechanisms with different phenotypes were selected, with E. coli ATCC 25922 as control strain. In addition, from each clinical isolate and control, a second strain was created, in which the SOS response was suppressed by deletion of the recA gene. Bacterial inocula from all 12 strains were then exposed to 1xMIC ciprofloxacin treatment (relative to the wild-type phenotype for each isogenic pair) for 1 h. Cell pellets were collected, and proteins were digested into peptides using trypsin. Protein identification and label-free quantification were done by liquid chromatography-mass spectrometry (LC–MS) in order to identify proteins that were differentially expressed upon deletion of recA in each strain. Data analysis and statistical analysis were performed using the MaxQuant and Perseus software. Results: The proteins with the lowest expression levels were: RecA (as control), AphA, CysP, DinG, DinI, GarL, PriS, PsuG, PsuK, RpsQ, UgpB and YebG; those with the highest expression levels were: Hpf, IbpB, TufB and RpmH. Most of these expression alterations were strain-dependent and involved DNA repair processes and nucleotide, protein and carbohydrate metabolism, and transport. In isolates with suppressed SOS response, the number of underexpressed proteins was higher than overexpressed proteins. Conclusion: High genomic and proteomic variability was observed among clinical isolates and was not associated with a specific resistant phenotype. This study provides an interesting approach to identify new potential targets to combat antimicrobial resistance.Artículo Characterizing Replisome Disassembly in Human Cells(Elsevier, 2024) Jones, Rebecca M.; Herrero Ruiz, Joaquín; caramuzza, Shaun; Nath, Sarmi; Liu, Chaoyu; Henklewska, Marta; Natsume, Toyoaki; Bristow, Robert G.; Romero Portillo, Francisco; Kanemaki, Masato T.; Gambus, Agnieszka; Universidad de Sevilla. Departamento de Microbiología; Wellcome Trust. UKTo ensure timely duplication of the entire eukaryotic genome, thousands of replication machineries (replisomes) act on genomic DNA at any time during S phase. In the final stages of this process, replisomes are unloaded from chromatin. Unloading is driven by polyubiquitylation of MCM7, a subunit of the terminated replicative helicase, and processed by p97/VCP segregase. Most of our knowledge of replication termination comes from model organisms, and little is known about how this process is executed and regulated in human somatic cells. Here we show that replisome disassembly in this system requires CUL2LRR1-driven MCM7 ubiquitylation, p97, and UBXN7 for unloading and provide evidence for “backup” mitotic replisome disassembly, demonstrating conservation of such mechanisms. Finally, we find that small-molecule inhibitors against Cullin ubiquitin ligases (CULi) and p97 (p97i) affect replisome unloading but also lead to induction of replication stress in cells, which limits their usefulness to specifically target replisome disassembly processes.Artículo Activity of Cefepime, Carbapenems and New β-lactam/β-lactamase Inhibitor Combinations on Enterobacter Cloacae Complex and Klebsiella Aerogenes in Spain (SMART 2016–2022)(Oxford University Press, 2024) Rodríguez Villodres, Ángel; Lepe Balsalobre, Esperanza; Ortiz de la Rosa, José Manuel; Giner Almaraz, Salvador; González De Herrero, Elisa; Cercenado, Emilia; García Fernández, Sergio; Benito, Rafael; Ponz Mir, Ricardo; Cantón, Rafael; Lepe Jiménez, José Antonio; Universidad de Sevilla. Departamento de Microbiología; Ministerio de Ciencia, Innovación y Universidades (MICINN). EspañaObjectives: To analyse the susceptibility profile to cefepime, carbapenems and new β-lactam/β-lactamase inhibitor combinations in Enterobacter cloacae complex and Klebsiella aerogenes isolated from intra-abdominal, urinary, respiratory and bloodstream infections in the SMART (Study for Monitoring Antimicrobial Resistance Trends) surveillance study in Spain. Methods: The susceptibilities of 759 isolates (473 E. cloacae complex and 286 K. aerogenes) collected in 11 Spanish hospitals from 2016 to 2022 were analysed following the EUCAST 2023 criteria. Molecular characterization looking for β-lactamase genes was performed through PCR and DNA sequencing analysis. Results: E. cloacae complex showed resistance to third-generation cephalosporins in 25% of the cases, whereas K. aerogenes was resistant in 35%. Regarding cefepime, resistance in E. cloacae was higher (10%) than in K. aerogenes (2%). Carbapenems showed >85% activity in both microorganisms. Ceftazidime/avibactam, imipenem/relebactam and meropenem/vaborbactam had good activity against these microorganisms (>95%). In contrast, the activity of ceftolozane/tazobactam was lower (80%). A high proportion of the isolates resistant to new β-lactam/β-lactamase inhibitor combinations carried a carbapenemase, mainly OXA-48-like and VIM-1. Conclusions: Ceftazidime/avibactam, imipenem/relebactam and meropenem/vaborbactam show high activity against both E. cloacae complex and K. aerogenes isolates recovered in the SMART-Spain study. In contrast, differences have been found in the case of cefepime, showing more activity against K. aerogenes than E. cloacae complex. These results are useful for antimicrobial stewardship programmes and for the implementation of local and national guidelines.
Artículo Survival of infection with TEM beta-lactamase-producing Escherichia coli with Pan-beta-lactam resistance(Elsevier, 2024) Rodriguez-Villodres, Angel; Ortiz de la Rosa, Jose Manuel; Galvez-Benitez, Lydia; Gascon, Maria Luisa; Penalva, German; Dorado Pardo, Francisco J.; Cisneros, José Miguel; Lepe Jiménez, José Antonio; Universidad de Sevilla. Departamento de Microbiología; Universidad de Sevilla. Departamento de MedicinaBackground: Antimicrobial resistance is a critical global health issue, significantly contributing to patient mortality. Recent antibiotic developments have aimed to counteract carbapenemase-producing Enterobacterales; however, the impact of their use on the emergence of antibiotic resistance is unknown. This study investigates the first case of a non-carbapenemase-producing, pan-β-lactam-resistant Escherichia coli strain from a patient previously treated with ceftolozane-tazobactam and cefiderocol. Methods: This study describes the clinical progression of a 39-year-old ICU patient who developed multiple infections, culminating in the isolation of a pan-β-lactam-resistant E. coli strain (EC554). The resistance profile was characterised through MIC determination, whole-genome sequencing, the use of the β-lactam inactivation method, RT-qPCR, efflux pump inhibition assays, outer membrane protein analysis, and blaTEM transformation. Findings: The EC554 isolate displayed resistance to all tested β-lactams and β-lactam-β-lactamase inhibitor combinations. Whole-genome sequencing revealed four plasmids in EC554, with the only β-lactamase gene being blaTEM-252 on the pEC554-PBR-X1-X1 plasmid. We found that the extremely resistant phenotype was attributable to a combination of different mechanisms: a high expression of TEM-252, efflux pump activity, porin loss, and PBP3 mutations. Interpretation: The findings illustrate the complex interplay of multiple resistance mechanisms in E. coli, highlighting the potential for high-level resistance even without carbapenemase production. This study underscores the importance of comprehensively characterising resistance mechanisms in order to inform effective treatment strategies and mitigate the spread of resistant strains.Artículo Effects of drought and increased temperature on phytochemical traits of the edible halophyte Crithmum maritimum: : Perspectives for future climatic scenarios(Elsevier, 2024-10) Martins Noguerol, Raquel; Rico Jiménez, Diego; Matías Resina, Luis; Pérez Ramos, Ignacio Manuel; Moreira, Xoaquín; Francisco, Marta; Álvarez Morales, Rosario; Gandullo Tovar, Jacinto Manuel; Moreno Pérez, Antonio Javier; Cambrollé Silva, Jesús; Universidad de Sevilla. Departamento de Biología Vegetal y Ecología; Universidad de Sevilla. Departamento de Microbiología; Agencia Estatal de Investigación. España; Ministerio de Ciencia e Innovación (MICIN). España; Junta de Andalucía; European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER); Universidad de SevillaClimate change, characterised by drought events and rising temperatures, exerts a significant threat to crop productivity and global food security. Halophytes, known for their resilience in harsh conditions, offer promising options for sustainable cultivation alternatives. Our study focused on Crithmum maritimum, commonly known as sea fennel, an edible halophyte with potential in the food and nutraceutical industries, to explore the impacts of drought and increased temperatures on its nutritional and antioxidant profiles. Different C. maritimum populations displayed high nutritional qualities, suitable for consumption despite appearing slight differences among localities. While both drought and increased temperatures affected plant growth and phytochemical profiles, their impact on nutritional value was minor. Surprisingly, drought induced an unexpected decline in phenolic content, challenging the assumption of increased antioxidants in response to water scarcity. Different rates of decrease in leaf production were observed among C. maritimum populations under drought, yet overall, they maintained similar levels, suggesting potential suitability for cultivation in environments with limited water availability. Diverse population-specific responses under climatic treatments revealed different alterations in amino acid and oxidative stress profiles, suggesting diverse adaptive strategies. These findings provide critical insights into C. maritimum adaptability to climate-driven changes, offering valuable information for future agricultural practices.Artículo Mpox in people with advanced HIV infection: a global case series(Elsevier, 2023-03-18) Mitja, O.; Alemany, A.; Marks, M.; Mora, J. I. L.; Rodríguez-Aldama, J. C.; Silva, M. S. T.; Herrera, E. A. C.; Crabtree-Ramírez, B.; Blanco, J. L.; Girometti, N.; Lepe Jiménez, José Antonio; Orkin, C. M.; SHARE-NET writing group; Universidad de Sevilla. Departamento de MicrobiologíaBackground People living with HIV have accounted for 38–50% of those affected in the 2022 multicountry mpox outbreak. Most reported cases were in people who had high CD4 cell counts and similar outcomes to those without HIV. Emerging data suggest worse clinical outcomes and higher mortality in people with more advanced HIV. We describe the clinical characteristics and outcomes of mpox in a cohort of people with HIV and low CD4 cell counts (CD4 <350 cells per mm3). Methods A network of clinicians from 19 countries provided data of confirmed mpox cases between May 11, 2022, and Jan 18, 2023, in people with HIV infection. Contributing centres completed deidentified structured case report sheets to include variables of interest relevant to people living with HIV and to capture more severe outcomes. We restricted this series to include only adults older than 18 years living with HIV and with a CD4 cell count of less than 350 cells per mm3 or, in settings where a CD4 count was not always routinely available, an HIV infection clinically classified as US Centers for Disease Control and Prevention stage C. We describe their clinical presentation, complications, and causes of death. Analyses were descriptive. Findings We included data of 382 cases: 367 cisgender men, four cisgender women, and ten transgender women. The median age of individuals included was 35 (IQR 30–43) years. At mpox diagnosis, 349 (91%) individuals were known to be living with HIV; 228 (65%) of 349 adherent to antiretroviral therapy (ART); 32 (8%) of 382 had a concurrent opportunistic illness. The median CD4 cell count was 211 (IQR 117–291) cells per mm3, with 85 (22%) individuals with CD4 cell counts of less than 100 cells per mm3 and 94 (25%) with 100–200 cells per mm3. Overall, 193 (51%) of 382 had undetectable viral load. Severe complications were more common in people with a CD4 cell count of less than 100 cells per mm3 than in those with more than 300 cells per mm3, including necrotising skin lesions (54% vs 7%), lung involvement (29% vs 0%) occasionally with nodules, and secondary infections and sepsis (44% vs 9%). Overall, 107 (28%) of 382 were hospitalised, of whom 27 (25%) died. All deaths occurred in people with CD4 counts of less than 200 cells per mm3. Among people with CD4 counts of less than 200 cells per mm3, more deaths occurred in those with high HIV viral load. An immune reconstitution inflammatory syndrome to mpox was suspected in 21 (25%) of 85 people initiated or re-initiated on ART, of whom 12 (57%) of 21 died. 62 (16%) of 382 received tecovirimat and seven (2%) received cidofovir or brincidofovir. Three individuals had laboratory confirmation of tecovirimat resistance. Interpretation A severe necrotising form of mpox in the context of advanced immunosuppression appears to behave like an AIDS-defining condition, with a high prevalence of fulminant dermatological and systemic manifestations and death.Artículo Soil microorganisms buffer the reduction in plant growth and physiological performance under combined abiotic stress in the halophyte Salicornia ramosissima(Elsevier, 2023-11-08) Mateos Naranjo, Enrique; Pérez Romero, Jesús Alberto; Puglielli, Giacomo; López Jurado, Javier; Mesa Marín, Jennifer; Pajuelo Domínguez, Eloísa; Rodríguez Llorente, Ignacio David; Redondo Gómez, Susana; Universidad de Sevilla. Departamento de Biología Vegetal y Ecología; Universidad de Sevilla. Departamento de Microbiología; Ministerio de Ciencia e Innovación (MICIN). España; Agencia Estatal de Investigación. España; European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER)The impact of multifactorial abiotic stress combinations on plant functional responses remains controversial, and general patterns of response are yet to emerge. This knowledge gap is particularly relevant for species with innate tolerance to environmental stress. Using the halophyte Salicornia ramosissima as a model species, we performed a multifactorial study with 16 experimental scenarios that included or not beneficial microorganisms in order to quantify their impact on plant growth, photosynthetic performance, osmotic adjustment and ion homeostasis. The experimental scenarios were characterized by the combination of four factors with two levels (salinity: 171 and 510 mM NaCl; water stress: yes and no; temperature min/max range: 14/25 and 18/29ºC and atmospheric CO2 concentration: 400 and 700 ppm). A plant growth-promoting rhizobacteria (PGPR) consortium was used as a proxy for positive biological interaction. The results revealed that the multifactorial stress combinations triggered unique functional responses, depending on the stress factors involved. However, there was an overall more negative impact on plant functional traits under the most extreme scenario (i.e., 510 mM NaCl + water stress + high temperature). Interestingly, the presence of PGPR was able to reverse this negative influence, although this effect was negligible under non-stressful conditions. Furthermore, the positive effect of PGPR was even magnified when coexisting with elevated atmospheric CO2 concentration. This response is associated with mitigation of the negative impacts of suboptimal factor combinations on plant growth, photosynthetic performance/efficiency, and water/nutrient homeostasis. Therefore, we conclude that the positive impact of microorganisms on halophyte tolerance in complex environmental matrices would only be determinant under extreme conditions in which plant intrinsic tolerance mechanisms would not be sufficient. Remarkably, this effect could be accentuated by increasing atmospheric CO2 concentration.Artículo Técnicas de biología molecular usadas en estudios publicados en revistas de medicina peruana, 2010-2019(Infomed, 2023) Pilco Ferreto, Nesstor; Medina Morillas, Carlos; Ponce Cusi, Richard; Universidad de Sevilla. Departamento de MicrobiologíaLas técnicas de biología molecular son fundamentales en medicina porque permiten el estudio a nivel molecular de enfermedades, el diagnóstico temprano, el desarrollo de terapias específicas y la medicina personalizada. Estas herramientas son esenciales para entender la base genética de las enfermedades y diseñar tratamientos precisos. El objetivo del estudio fue describir la cantidad y tipos de artículos que utilizaron técnicas de biología molecular, publicados en cuatro revistas de medicina peruana durante los años 2010 y 2019. En este trabajo se describieron 1415 artículos en cuatro revistas médicas peruanas entre artículos originales y reportes de casos. A pesar de un aumento en los reportes de casos sin técnicas de biología molecular, no hubo diferencias significativas entre artículos originales y reportes de casos. Las técnicas más comunes fueron: PCR, ELISA y electroforesis. Los hospitales e institutos de salud lideraron en cantidad de publicaciones. Estos resultados proporcionan valiosos datos para futuros estudios y perspectivas sobre el uso de estas técnicas en la producción científica médica peruana.Artículo The Gap Junction Inhibitor Octanol Decreases Proliferation and Increases Glial Differentiation of Postnatal Neural Progenitor Cells(Multidisciplinary Digital Publishing Institute (MDPI), 2024-06-07) Talaverón Aguilocho, Rocío; Morado Díaz, Camilo José; Herrera Delgado, Alejandro; Gálvez, Victoria; Pastor Loro, Ángel Manuel; Rodríguez Matarredona, Esperanza; Universidad de Sevilla. Departamento de Fisiología; Universidad de Sevilla. Departamento de Microbiología; Ministerio de Ciencia e Innovación (MICIN). España; Junta de AndalucíaNeural precursor cells (NPCs) that persist in the postnatal/adult subventricular zone (SVZ) express connexins that form hemichannels and gap junctions. Gap junctional communication plays a role in NPC proliferation and differentiation during development, but its relevance on postnatal age remains to be elucidated. In this work we aimed to evaluate the effect of the blockade of gap junctional communication on proliferation and cell fate of NPCs obtained from the SVZ of postnatal rats. NPCs were isolated and expanded in culture as neurospheres. Electron microscopy revealed the existence of gap junctions among neurosphere cells. Treatment of cultures with octanol, a broad-spectrum gap junction blocker, or with Gap27, a specific blocker for gap junctions formed by connexin43, produced a significant decrease in bromodeoxyuridine incorporation. Octanol treatment also exerted a dose-dependent antiproliferative effect on glioblastoma cells. To analyze possible actions on NPC fate, cells were seeded in the absence of mitogens. Treatment with octanol led to an increase in the percentage of astrocytes and oligodendrocyte precursors, whereas the percentage of neurons remained unchanged. Gap27 treatment, in contrast, did not modify the differentiation pattern of SVZ NPCs. Our results indicate that general blockade of gap junctions with octanol induces significant effects on the behavior of postnatal SVZ NPCs, by reducing proliferation and promoting glial differentiation.Artículo First Steps Towards a Vaccine against Acinetobacter baumannii(Bentham Science, 2013) García Quintanilla, Meritxell de Jesús; Pulido, Marina R.; McConnell, Michael J.; Universidad de Sevilla. Departamento de Microbiología; Universidad de Sevilla. CTS210: Resistencia a AntimicrobianosAcinetobacter baumannii has become an important cause of human infections, most notably in the hospital setting. In addition, the global dissemination of multidrug resistant strains has complicated effective antibiotic therapy of infections produced by this pathogen, necessitating the development of novel treatment and prevention strategies. Active and passive immunization approaches have begun to be explored in experimental animal models as potential alternative therapies for A. baumannii. In the present review, we discuss the advantages and disadvantages of each therapeutic strategy with respect to A. baumannii infections, and summarize the recent studies that have explored these approaches. The single antigen candidates that have been tested include, the outer membrane protein OmpA, the membrane transporter Ata, the biofilm-associated protein Bap, the K1 capsular polysaccharide and the membrane associated polysaccharide poly-N-acetyl-β -(1-6)-glucosamine. Strategies employing multicomponent antigens include inactivated whole cells, outer membrane complexes and outer membrane vesicles. The strengths and limitations of each approach are discussed and the challenges that remain to be addressed for successful A. baumannii vaccine development are highlighted.Artículo A lipopolysaccharide-free outer membrane vesicle vaccine protects against Acinetobacter baumannii infection(Elsevier, 2020-01) Pulido, Marina R.; García Quintanilla, Meritxell de Jesús; Pachón Díaz, Jerónimo; McConnell, Michael J.; Universidad de Sevilla. Departamento de Microbiología; Universidad de Sevilla. Departamento de Medicina; Universidad de Sevilla. CTS210: Resistencia a Antimicrobianos; Universidad de Sevilla. CTS203: Estudio de las Enfermedades InfecciosasOuter membrane vesicles (OMVs) were isolated from an Acinetobacter strain deficient in lipopolysaccharide (LPS) due to a mutation in lpxD (IB010). Two immunizations with 10 µg of IB010 OMVs elicited total IgG, IgM, IgG1 and IgG2c titers similar to those observed after immunization with OMVs derived from the parental strain (ATCC 19606), and IB010 OMVs plus purified LPS. Immunization with IB010 OMVs resulted in significantly reduced post-infection spleen bacterial loads and serum IL-1β and IL-6 levels compared to control mice in a disseminated sepsis model. Mice immunized with 10 µg IB010 OMVs demonstrated significant, but partial, protection (75%) against infection, whereas mice immunized with ATCC 19606 OMVs or IB010 OMVs plus purified LPS were completely protected. Immunization of mice with 100 µg of IB010 OMVs completely protected mice from infection. This study demonstrates that LPS deficient A. baumannii produces OMVs, and that immunization with these OMVs elicits protective immunity against infection.Artículo Identification of a Stable Chromosomal Tandem Multicopy of bla(VIM-63), a New bla(VIM-2) Carbapenemase(American Society for Microbiology, 2022-07) Pulido, Marina R.; García Montaner, Andrea; López Cerero, Lorena; Fernández Cuenca, Felipe Manuel; Gutiérrez-Fernández, José; Pascual Hernández, Álvaro; Universidad de Sevilla. Departamento de Microbiología; Universidad de Sevilla. CTS210: Resistencia a AntimicrobianosThis study characterizes a new genetic structure containing a multicopy of a bla(VIM-2) variant with an A676C substitution, bla(VIM-63). This gene was detected on the chromosome of two carbapenem-resistant clinical strains of Citrobacter freundii ST22 recovered from two patients, separated by a 6-month period, and previously in Pseudomonas aeruginosa ST2242 from the same hospital unit. Short-read sequencing was used to characterize the new variant in both species, and long-read sequencing was used to characterize the genome of C. freundii. On the P. aeruginosa chromosome, the bla(VIM-63) gene was inserted between ISPsy 42-type sequences, flanked by an intl1 sequence, nearby aph(3′)-VI, and sul1. On the C. freundii chromosome, the bla(VIM-63) gene was inserted into a Tn6230-like transposon as a stable five-tandem-repeat multimer, flanked by the same intl1 as in P. aeruginosa. This structure was stable across subcultures and did not change in the presence of carbapenems. The bla(VIM-63) gene was cloned into the pCR-Blunt plasmid to study antimicrobial susceptibility patterns and into pET29a for kinetic activity analysis. VIM-63 showed higher K(m) values than VIM-2 for ceftazidime and cefepime and higher k(cat) values for cefotaxime, ceftazidime, imipenem, and ertapenem, without differences in MIC values. This is the first study to describe this new variant, VIM-63, in two different species with a chromosomal location integrated into different mobile elements and the first to describe a stable multimer of a metallo-β-lactamase. Despite the amino acid substitution, the susceptibility pattern of the new variant was similar to that of VIM-2. IMPORTANCE: VIM group metallo-β-lactamases are usually captured by IntI1 integrases. This work describes the detection for the first time of a novel, previously unknown variant of VIM-2, VIM-63. This carbapenemase has been found on the chromosome of two different species, Citrobacter freundii and Pseudomonas aeruginosa, from the same hospital. The adjacent genetic environment of the bla(VIM-63) gene would indicate that the capture of this gene by IntI1 has occurred in two different genetic events in each of the species, and in one there has been a stable integration of tandem copies of this gene.Artículo Characterization of Extended-Spectrum β-Lactamase-Producing Shigella sonnei in Spain: Expanding the Geographic Distribution of Sequence Type 152/CTX-M-27 Clone(American Society for Microbiology, 2022-07) López Cerero, Lorena; Stolz, E.; Pulido, Marina R.; Pascual Hernández, Álvaro; Universidad de Sevilla. Departamento de Microbiología; Universidad de Sevilla. CTS210: Resistencia a AntimicrobianosWe describe the first occurrence in Spain of community cases of CTX-M-27-producing Shigella sonnei sequence type 152 (ST152), resistant to quinolones and azithromycin. The cases included adult males and also one pediatric case. The isolates were clustered together with an Australian isolate and differed from other outbreak-causing strains in England by more than 50 alleles. They carried the blaCTX-M-27 gene on an 83-Kb F2:A-:B- plasmid, similar to that found in a British isolate.Artículo Synergistic effect of SOS response and GATC methylome suppression on antibiotic stress survival in Escherichia coli(https://journals.asm.org/doi/epdf/10.1128/aac.01392-22, 2023-03) Díaz Díaz, S.; Recacha, E.; Pulido, Marina R.; Romero Muñoz, María; De Gregorio Iaria, Belén; Docobo Pérez, Fernando; Pascual Hernández, Álvaro; Rodríguez Martínez, José Manuel; Universidad de Sevilla. Departamento de Microbiología; Universidad de Sevilla. CTS210: Resistencia a AntimicrobianosThe suppression of the SOS response has been shown to enhance the in vitro activity of quinolones. Furthermore, Dam-dependent base methylation has an impact on susceptibility to other antimicrobials affecting DNA synthesis. Here, we investigated the interplay between these two processes, alone and in combination, in terms of antimicrobial activity. A genetic strategy was used employing single- and double-gene mutants for the SOS response (recA gene) and the Dam methylation system (dam gene) in isogenic models of Escherichia coli both susceptible and resistant to quinolones. Regarding the bacteriostatic activity of quinolones, a synergistic sensitization effect was observed when the Dam methylation system and the recA gene were suppressed. In terms of growth, after 24 h in the presence of quinolones, the Δdam ΔrecA double mutant showed no growth or delayed growth compared to the control strain. In bactericidal terms, spot tests showed that the Δdam ΔrecA double mutant was more sensitive than the ΔrecA single mutant (about 10- to 10² -fold) and the wild type (about 10³- to 10^4-fold) in both susceptible and resistant genetic backgrounds. Differences between the wild type and the Δdam ΔrecA double mutant were confirmed by time-kill assays. The suppression of both systems, in a strain with chromosomal mechanisms of quinolone resistance, prevents the evolution of resistance. This genetic and microbiological approach demonstrated the enhanced sensitization of E. coli to quinolones by dual targeting of the recA (SOS response) and Dam methylation system genes, even in a resistant strain model.