María Guembe
@iisgm.com
Clinical Microbiology and Infectious Diseases Department, Gregorio Marañón Hospital
Fundación para la Investigación Biomédica del Hospital Gregorio Marañón
RESEARCH, TEACHING, or OTHER INTERESTS
Microbiology (medical), Orthopedics and Sports Medicine, Microbiology, Infectious Diseases
Scopus Publications
Scopus Publications
Enea Gino Di Domenico, Alessandra Oliva, and María Guembe
MDPI AG
In this Special Issue, titled “Biofilm-Related Infections in Healthcare”, we have reported considerable progress in understanding the physiology and pathology of biofilms [...]
M. Pérez-Granda, Álvaro Irigoyen-von-Sierakowski, Neera Toledo, Eva Rodríguez, María Luisa Cruz, Giovanna Hernanz, José Antonio Serra, M. Kestler, Patricia Muñoz and M. Guembe
Marta Díaz-Navarro, Rama Hafian, María Jesús Pérez-Granda, Emilia Cercenado, Patricia Muñoz, and María Guembe
Elsevier BV
Antonio Benjumea, Marta Díaz-Navarro, Rama Hafian, Emilia Cercenado, Mar Sánchez-Somolinos, Javier Vaquero, Francisco Chana, Patricia Muñoz, and María Guembe
Frontiers Media SA
Marta Díaz-Navarro, Álvaro Irigoyen Von-Sierakowski, María Palomo, Pilar Escribano, Jesús Guinea, Almudena Burillo, Alicia Galar, Patricia Muñoz, and María Guembe
Elsevier BV
Miguel Márquez-Gómez, Marta Díaz-Navarro, Andrés Visedo, Rama Hafian, José Matas, Patricia Muñoz, Javier Vaquero, María Guembe, and Pablo Sanz-Ruíz
MDPI AG
Irrigation and debridement using an irrigation solution is a fundamental step during the surgical treatment of both acute and chronic periprosthetic joint infection (PJI). However, there is no consensus on the optimal solution, nor is there sufficient evidence on the optimal irrigation time and combination of solutions. Therefore, it is necessary to determine which solution or combination of solutions is most efficacious against biofilm, as well as the optimal irrigation time. We conducted an experimental in vitro model by inoculating stainless steel discs with ATCC strains of methicillin-susceptible Staphylococcus aureus, methicillin-resistant S. aureus, Pseudomonas aeruginosa, and a clinical strain of Staphylococcus epidermidis. The discs were all irrigated with commonly used antiseptic solutions (10% and 3% povidone iodine, hydrogen peroxide, 3% acetic acid, and Bactisure™) for 1 min, 3 min, and 5 min and their combinations for 9 min (3 min each) vs. sterile saline as a positive control. We evaluated the reduction in biofilm based on colony-forming unit (cfu) counts and in combination assays, also based on cell viability and scanning electron microscopy. All antiseptics alone reduced more than 90% of cfu counts after 1 min of irrigation; the worst results were for hydrogen peroxide and 3% acetic acid. When solutions were sequentially combined, the best results were observed for all those starting with acetic acid, in terms of both reduction of log cfu/mL counts and viable cells. We consider that a combination of antiseptic solutions, particularly that comprising the sequence acetic acid + povidone iodine + hydrogen peroxide, would be the best option for chemical debridement during PJI surgery.
Mar Sánchez-Somolinos, Marta Díaz-Navarro, Antonio Benjumea, José Matas, Javier Vaquero, Patricia Muñoz, Pablo Sanz-Ruíz, and María Guembe
MDPI AG
Based on previous studies by our group in which we demonstrated that dalbavancin loaded in bone cement had good elution capacity for the treatment of biofilm-related periprosthetic infections, we now assess the anti-biofilm activity of dalbavancin and compare it with that of vancomycin over a 3-month period. We designed an in vitro model in which we calculated the percentage reduction in log cfu/mL counts of sonicated steel discs contaminated with staphylococci and further exposed to bone cement discs loaded with 2.5% or 5% vancomycin and dalbavancin at various timepoints (24 h, 48 h, 1 week, 2 weeks, 6 weeks, and 3 months). In addition, we tested the anti-biofilm activity of eluted vancomycin and dalbavancin at each timepoint based on a 96-well plate model in which we assessed the percentage reduction in metabolic activity. We observed a significant decrease in the dalbavancin concentration from 2 weeks of incubation, with sustained anti-biofilm activity up to 3 months. In the case of vancomycin, we observed a significant decrease at 1 week. The concentration gradually increased, leading to significantly lower anti-biofilm activity. The percentage reduction in cfu/mL counts was higher for dalbavancin than for vancomycin at both the 2.5% and the 5% concentrations. The reduction in log cfu/mL counts was higher for S. epidermidis than for S. aureus and was particularly more notable for 5% dalbavancin at 3 months. In addition, the percentage reduction in metabolic activity also decreased at 3 months in 5% dalbavancin and 5% vancomycin, with more notable values recorded for the latter.
David Rodríguez-Temporal, Rafael Díez, Marta Díaz-Navarro, Pilar Escribano, Jesús Guinea, Patricia Muñoz, Belén Rodríguez-Sánchez, and María Guembe
Frontiers Media SA
BackgroundThe traditional method for assessing the capacity of a microorganism to produce biofilm is generally a static in vitro model in a multi-well plate using the crystal violet (CV) binding assay, which takes 96 h. Furthermore, while the method is simple to perform, its reproducibility is poor.ObjectiveWe evaluated whether matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) could make it possible to differentiate between high-and low-biofilm-producing microorganisms on 24-h cultures of Staphylococcus aureus and Candida albicans.MethodsWe included 157 strains of S. aureus and 91 strains of C. albicans obtained from the blood cultures of patients with bacteremia/candidemia. We tested biofilm production using the CV binding assay as the gold standard to classify strains as low or high biofilm producers. We then applied MALDI-TOF MS to create a machine learning–based predictive model using 40 strains of S. aureus and C. albicans, each with extreme absorbance values, and validated this approach with the remaining 117 and 51 strains using the random forest algorithm and the support vector machine algorithm, respectively.ResultsOverall, 81.2% of the S. aureus strains (95/117) and 74.5% of the C. albicans strains (38/51) used for validation were correctly categorized, respectively, as low and high-biofilm-producing.ConclusionClassification based on MALDI-TOF MS protein spectra enables us to predict acceptable information about the capacity of 24-h cultures of S. aureus and C. albicans to form biofilm.
Marta Díaz-Navarro, Rafael Samaniego, Juan Carlos Piqueras, Rafael Díez, Rama Hafian, Irene Manzano, Patricia Muñoz, and María Guembe
Frontiers Media SA
BackgroundThe differential time to positivity (DTTP) technique is recommended for the conservative diagnosis of catheter-related bloodstream infection (C-RBSI). The technique is based on a 120-minute difference between microbial growth in blood drawn through the catheter and blood drawn through a peripheral vein. However, this cut-off has failed to confirm C-RBSI caused by Candida spp. and Staphylococcus aureus.ObjectiveWe hypothesized that the biofilm of both microorganisms disperses faster than that of other microorganisms and that microbial load is rapidly equalized between catheter and peripheral blood. Therefore, our aim was to compare the biofilm dynamics of various microorganisms.MethodsBiofilm of ATCC strains of methicillin-resistant Staphylococcus epidermidis, methicillin-susceptible S. aureus, Enterococcus faecalis, Escherichia coli and Candida albicans was grown on silicon disks and analyzed using time-lapse optical microscopy. The time-lapse images of biofilms were processed using ImageJ2 software. Cell dispersal time and biofilm thickness were calculated.ResultsThe mean (standard deviation) dispersal time in C. albicans and S. aureus biofilms was at least nearly 3 hours lower than in biofilm of S. epidermidis, and at least 15 minutes than in E. faecalis and E. coli biofilms.ConclusionOur findings could explain why early dissemination of cells in C. albicans and S. aureus prevents us from confirming or ruling out the catheter as the source of the bloodstream infection using the cut-off of 120 minutes in the DTTP technique. In addition, DTTP may not be sufficiently reliable for E. coli since their dispersion time is less than the cut-off of 120 minutes.
Mar Sánchez-Somolinos, Marta Díaz-Navarro, Antonio Benjumea, Marta Tormo, José Matas, Javier Vaquero, Patricia Muñoz, Pablo Sanz-Ruíz, and María Guembe
MDPI AG
Antibiotic-loaded bone cement is the most widely used approach for the treatment of biofilm-induced septic sequelae in orthopedic surgery. Dalbavancin is a lipoglycopeptide that acts against Gram-positive bacteria and has a long half-life, so we aimed to assess whether it could be a new alternative drug in antibiotic-loaded bone cement for the treatment of periprosthetic joint infections. We assessed the elution capacity of dalbavancin and compared it with that of vancomycin in bone cement. Palacos®R (Heraeus Medical GmbH, Wehrheim, Germany) bone cement was manually mixed with each of the antibiotics studied at 2.5% and 5%. Three cylinders were obtained from each of the mixtures; these were weighed and incubated in 5 mL phosphate-buffered saline at 37°C under shaking for 1 h, 2 h, 4 h, 8 h, 24 h, 48 h, 168 h, and 336 h. PBS was replenished at each time point. The samples were analyzed using high-performance liquid chromatography (vancomycin) and mass cytometry (dalbavancin). Elution was higher than the minimum inhibitory concentration (MIC)90 for both antibiotics after 14 days of study. The release of vancomycin at 14 days was higher than of dalbavancin at each concentration tested (p = 0.05, both). However, the cumulative release of 5% dalbavancin was similar to that of 2.5% vancomycin (p = 0.513). The elution capacity of dalbavancin reached a cumulative concentration similar to that of vancomycin. Moreover, considering that the MIC90 of dalbavancin is one third that of vancomycin (0.06 mg/L and 2 mg/L, respectively) and given the long half-life of dalbavancin, it may be a new alternative for the treatment of biofilm-related periprosthetic infections when loaded in bone cement.
Michele Castellano, Mercedes Marín, Luis Alcalá, Iskra Cunnas, Belén Rodríguez, Mª Jesús Ruíz, José Lasso, Rosa Pérez-Cano, Emilio Bouza, Patricia Muñoz,et al.
Elsevier BV
Enea Gino Di Domenico, Alessandra Oliva, and María Guembe
MDPI AG
Biofilm is the trigger for the majority of infections caused by the ability of microorganisms to adhere to tissues and medical devices. Microbial cells embedded in the biofilm matrix are highly tolerant to antimicrobials and escape the host immune system. Thus, the refractory nature of biofilm-related infections (BRIs) still represents a great challenge for physicians and is a serious health threat worldwide. Despite its importance, the microbiological diagnosis of a BRI is still difficult and not routinely assessed in clinical microbiology. Moreover, biofilm bacteria are up to 100–1000 times less susceptible to antibiotics than their planktonic counterpart. Consequently, conventional antibiograms might not be representative of the bacterial drug susceptibility in vivo. The timely recognition of a BRI is a crucial step to directing the most appropriate biofilm-targeted antimicrobial strategy.
Antonio Benjumea, Marta Díaz-Navarro, Rama Hafian, Emilia Cercenado, Mar Sánchez-Somolinos, Javier Vaquero, Francisco Chana, Patricia Muñoz, and María Guembe
Frontiers Media SA
BackgroundTranexamic acid (TXA) is an antifibrinolytic agent applied in orthopedic surgery and has been proven to reduce post-surgery infection rates. We previously showed that TXA also had an additional direct antimicrobial effect against planktonic bacteria. Therefore, we aimed to evaluate whether it has a synergistic effect if in combination with antibiotics.Materials and MethodsThree ATCC and seven clinical strains of staphylococci were tested against serial dilutions of vancomycin and gentamicin alone and in combination with TXA at 10 and 50 mg/ml. The standardized microtiter plate method was used. Minimal inhibitory concentrations (MICs) were calculated by standard visualization of well turbidity (the lowest concentration at which complete absence of well bacterial growth was observed by the researcher) and using the automated method (the lowest concentration at which ≥80% reduction in well bacterial growth was measured using a spectrophotometer).ResultsTranexamic acid-10 mg/ml reduced the MIC of vancomycin and gentamicin with both the standard method (V: 1-fold dilution, G: 4-fold dilutions) and the automated turbidity method (vancomycin: 8-fold dilutions, gentamicin: 8-fold dilutions). TXA-50 mg/ml reduced the MIC of gentamicin with both the standard turbidity method (6-fold dilutions) and the automated turbidity method (1-fold dilutions). In contrast, for vancomycin, the MIC remained the same using the standard method, and only a 1-fold dilution was reduced using the automated method.ConclusionOurs was a proof-of-concept study in which we suggest that TXA may have a synergistic effect when combined with both vancomycin and gentamicin, especially at 10 mg/ml, which is the concentration generally used in clinical practice.
Marta Díaz-Navarro, Rama Hafian, Irene Manzano, María J. Pérez-Granda, Emilia Cercenado, Cristina Pascual, Carmen Rodríguez, Patricia Muñoz, and María Guembe
Springer Science and Business Media LLC
María Consuelo Latorre, Luis Alcalá, Michele Castellano, José Lasso, Rosa Pérez-Cano, Emilio Bouza, Patricia Muñoz, and María Guembe
Elsevier BV
Francesca Sivori, Ilaria Cavallo, Daniela Kovacs, Maria Guembe, Isabella Sperduti, Mauro Truglio, Martina Pasqua, Grazia Prignano, Arianna Mastrofrancesco, Luigi Toma,et al.
American Society for Microbiology
Staphylococcus aureus is the most common cause of skin and soft tissue infections (SSTIs) worldwide. In addition, methicillin-resistant S. aureus (MRSA) is increasingly frequent in postoperative infections and responsible for a large number of hospital readmissions and deaths.
Marta Díaz-Navarro, Rama Hafian, Irene Manzano, María J. Pérez-Granda, Emilia Cercenado, Cristina Pascual, Carmen Rodríguez, Patricia Muñoz, and María Guembe
Springer Science and Business Media LLC
Borja Fernández-Ibarburu, Marta Díaz-Navarro, Gorka Ibarra, Andrés Rivera, Rama Hafian, Ãlvaro Irigoyen, Raquel Carrillo, Rosa Pérez-Cano, Patricia Muñoz, Ángela García-Ruano,et al.
Frontiers Media SA
BackgroundIn the practice of breast augmentation and reconstruction, implant irrigation with various solutions has been widely used to prevent infection and capsular contracture, but to date, there is no consensus on the optimal protocol to use. Recently, application of povidone iodine (PI) for 30 min has shown in vitro to be the most effective irrigating formula in reducing contamination in smooth breast implants. However, as 30 min is not feasible intraoperatively, it is necessary to determine whether shorter times could be equally effective as well as to test it in both smooth and textured implants.MethodsWe tested the efficacy of 10% PI at 1′, 3′, and 5′ against biofilms of 8 strains (2 ATCC and 6 clinical) of Staphylococcus spp. on silicone disks obtained from Mentor® and Polytech® implants of different textures. We analyzed the percentage reduction of cfu counts, cell viability and bacterial density between treatment (PI) and control (sterile saline, SS) groups for each time of application. We consider clinical significance when > 25% reduction was observed in cell viability or bacterial density.ResultsAll textured implants treated with PI at any of the 3 exposure times reduced 100% bacterial load by culture. However, none of the implants reached enough clinical significance in percentage reduction of living cells. Regarding bacterial density, only 25–50 μm Polytxt® Polytech® implants showed significant reduction at the three PI exposure times.ConclusionPI is able to inhibit bacterial growth applied on the surface of breast implants regardless of the exposure time. However, no significant reduction on living cells or bacterial density was observed. This lack of correlation may be caused by differences in texture that directly affect PI absorption.
María Consuelo Latorre, Beatriz Alonso, Raquel Cruces, Aurora Sanz, Patricia Muñoz, and María Guembe
Elsevier BV
Antonio Benjumea, Marta Díaz-Navarro, Rama Hafian, Mar Sánchez-Somolinos, Javier Vaquero, Francisco Chana, Patricia Muñoz, and María Guembe
American Society for Microbiology
The possible use of TXA as an antibiotic agent in addition to its antifibrinolytic effect may play an important role in the prevention of prosthetic joint infection.
Fiorentina Ascenzioni, Axel Cloeckaert, Enea Gino Di Domenico, Catherine Dunyach-Remy, and María Guembe
Frontiers Media SA
HAL is a multi-disciplinary open access archive for the deposit and dissemination of scientific research documents, whether they are published or not. The documents may come from teaching and research institutions in France or abroad, or from public or private research centers. L’archive ouverte pluridisciplinaire HAL, est destinée au dépôt et à la diffusion de documents scientifiques de niveau recherche, publiés ou non, émanant des établissements d’enseignement et de recherche français ou étrangers, des laboratoires publics ou privés.
María Consuelo Latorre, María Jesús Pérez-Granda, Paul B Savage, Beatriz Alonso, Pablo Martín-Rabadán, Rafael Samaniego, Emilio Bouza, Patricia Muñoz, and María Guembe
Oxford University Press (OUP)
Abstract Background Ventilator-associated pneumonia is one of the most common nosocomial infections, caused mainly by bacterial/fungal biofilm. Therefore, it is necessary to develop preventive strategies to avoid biofilm formation based on new compounds. Objectives We performed an in vitro study to compare the efficacy of endotracheal tubes (ETTs) coated with the ceragenin CSA-131 and that of uncoated ETTs against the biofilm of clinical strains of Pseudomonas aeruginosa (PA), Escherichia coli (EC) and Staphylococcus aureus (SA). Methods We applied an in vitro bench top model using coated and uncoated ETTs that were treated with three different clinical strains of PA, EC and SA for 5 days. After exposure to biofilm, ETTs were analysed for cfu count by culture of sonicate and total number of cells by confocal laser scanning microscopy. Results The median (IQR) cfu/mL counts of PA, EC and SA in coated and uncoated ETTs were, respectively, as follows: 1.00 × 101 (0.0–3.3 × 102) versus 3.32 × 109 (6.6 × 108–3.8 × 109), P < 0.001; 0.0 (0.0–5.4 × 103) versus 1.32 × 106 (2.3 × 103–5.0 × 107), P < 0.001; and 8.1 × 105 (8.5 × 101–1.4 × 109) versus 2.7 × 108 (8.6 × 106–1.6 × 1011), P = 0.058. The median (IQR) total number of cells of PA, EC and SA in coated and non-coated ETTs were, respectively, as follows: 11.0 [5.5–not applicable (NA)] versus 87.9 (60.5–NA), P = 0.05; 9.1 (6.7–NA) versus 62.6 (42.0–NA), P = 0.05; and 97.7 (94.6–NA) versus 187.3 (43.9–NA), P = 0.827. Conclusions We demonstrated significantly reduced biofilm formation in coated ETTs. However, the difference for SA was not statistically significant. Future clinical studies are needed to support our findings.
María Jesús Pérez-Granda, José María Barrio, Raquel Cruces, Beatriz Alonso, Pablo Martín-Rabadán, Inmaculada Collado, and María Guembe
Elsevier BV
Beatriz Alonso, María Jesús Pérez-Granda, María Consuelo Latorre, Carlos Sánchez-Carrillo, Emilio Bouza, Patricia Muñoz, and María Guembe
Elsevier BV
Beatriz Alonso, Laia Fernández-Barat, Enea Gino Di Domenico, Mercedes Marín, Emilia Cercenado, Irene Merino, Manuela de Pablos, Patricia Muñoz, and María Guembe
Springer Science and Business Media LLC
An amendment to this paper has been published and can be accessed via the original article.