Uropathogenic bacteria and deductive genomics towards antimicrobial resistance, virulence, and potential drug targets / Bacterias uropatógenas y genómica deductiva hacia la resistencia, virulencia y posibles objetivos farmacológicos a los antimicrobianos

Urinary tract infections (UTIs) are among the most prevalent bacterial infections affecting people in inpatient and outpatient settings. The current study aimed to sequence the genome of uropathogenic Escherichia coli strain CUI-B1 resourced from a woman having uncomplicated cystitis and pyelonephritis. Followed by deductive genomics towards potential drug targets using E. coli strain CUI-B1, strain O25b: H4-ST131, Proteus mirabilis strain HI4320, Klebsiella pneumoniae strain 1721, and Staphylococcus saprophyticus strain ATCC 15305 uropathogenic strains. Comparative genome analysis revealed that genes related to the survival of E. coli, P. mirabilis, K. pneumoniae, and S. saprophyticus, such as genes of metal-requiring proteins, defense-associated genes, and genes associated with general physiology, were found to be highly conserved in the genomes including strain CUI-B1. However, the genes responsible for virulence and drug resistance, mainly those that are involved in bacterial secretion, fimbriae, adherence, and colonization, were found in various genomic regions and varied from one species to another or within the same species. Based on the genome sequence, virulence, and antimicrobial-resistant gene dataset, the subtractive proteomics approach revealed 22 proteins mapped to the pathogen’s unique pathways and among them, entB, clbH, chuV, and ybtS were supposed to be potential drug targets and the single drug could be utilized for all above-mentioned strains. These results may provide the foundation for the optimal target for future discovery of drugs for E. coli-, P. mirabilis-, K. pneumoniae-, and S. saprophyticus-based infections and could be investigated further to employ in personalized drug development.(AU)

Rapid AST: Possibility of inferring resistance mechanisms with complex phenotypes / Pruebas rápidas de sensibilidad a los antimicrobianos: ¿es posible inferir mecanismos de resistencia con fenotipos complicados?

The new automated systems designed for rapid performance of AST have significantly reduced the response time for susceptibility testing of microorganisms causing bacteremia and sepsis. The Accelerate Pheno® system (AAC) is one such system. Our objective for this study was to determine whether the AAC system is capable of providing an accurate susceptibility profile to infer resistance mechanisms in different carbapenemase-producing isolates when compared to the MicroScan WalkAway System (MWS). Disk diffusion method was also performed on all isolates as a reference method. Additionally, we compared the results obtained with the routine AST production system. We selected 19 isolates from the cryobank of the Microbiology department, all of which were carbapenemase-producing gram-negative bacilli. AAC was able to identify and infer the resistance of a total of 10 isolates, with an EA and CA of 84.2% for meropenem and 88.2% and 64.7% for ertapenem EA and CA, respectively. If we consider the disk diffusion technique, the CA was 57.9% and 76.5% for meropenem and ertapenem. However, in the presence of carbapenemases, AAC was not able to provide adequate MICs or infer the resistance mechanisms of the isolates accurately. Further studies with a larger number of isolates, including the new antibiotics ceftolozane/tazobactam and ceftazidime/avibactam, are needed for a more comprehensive comparison. (AU)

Los nuevos sistemas automatizados diseñados para la realización rápida de antibiogramas han reducido significativamente el tiempo de respuesta para las pruebas de susceptibilidad de los microorganismos causantes de bacteriemia y sepsis. El sistema Accelerate Pheno® (AAC) es uno de ellos. Nuestro objetivo para este estudio era determinar si el sistema AAC es capaz de proporcionar un perfil de sensibilidad preciso para inferir mecanismos de resistencia en diferentes aislados productores de carbapenemasas en comparación con el sistema MicroScan WalkAway (MWS). El método de disco difusión fue incluido también en todos los aislados como método de referencia. Además, comparamos los resultados obtenidos con el sistema rutinario de producción de antibiogramas rápidos. Seleccionamos 19 aislados del criobanco del departamento de Microbiología, todos ellos bacilos gramnegativos productores de carbapenemasas. AAC fue capaz de identificar e inferir la resistencia de un total de 10 aislados, con una EA y CA del 84,2% para el meropenem y del 88,2% y 64,7% para la EA y CA del ertapenem, respectivamente. Si consideramos la técnica de disco difusión, la CA fue de un 57.9% y de un 76.5% para meropenem y ertapenem. Sin embargo, en presencia de carbapenemasas, AAC no fue capaz de proporcionar CMIs adecuadas ni de inferir con precisión los mecanismos de resistencia de los aislados. Se necesitan más estudios con un mayor número de aislados incluyendo también los nuevos antibióticos ceftolozano/tazobactam y ceftazidima/avibactam para una comparación más exhaustiva. (AU)

Narcolepsia multirresistente / Multidrug resistant narcolepsy

Introducción: La narcolepsia de tipo 1 es una enfermedad degenerativa focal del hipotálamo que afecta selectivamente a las neuronas productoras de orexina (hipocretina). Presenta múltiples manifestaciones clínicas, tanto en vigilia como en sueño. Con frecuencia, los síntomas son tan disruptivos que ocasionan enorme sufrimiento y deterioro de la calidad de vida de los pacientes. Aunque en ocasiones es suficiente con un abordaje no farmacológico, la gran mayoría de los enfermos necesita medicación para un adecuado control clínico. Caso clínico: Varón que a los 43 años comenzó a presentar de forma aguda excesiva somnolencia diurna y episodios de cataplejía. Tras un exhaustivo estudio se le diagnosticó narcolepsia de tipo 1. A lo largo de la evolución de la enfermedad se le prescribieron antidepresivos, neuroestimulantes y oxibato sódico, en monoterapia o en combinación. La respuesta al tratamiento farmacológico fue insuficiente y se acompañó de numerosos efectos secundarios. Tras la introducción de pitolisant se objetivó una franca mejoría de los síntomas, y se consiguió reducir la dosis de los otros fármacos y de sus efectos adversos. Conclusión: Son numerosas las medidas disponibles en la actualidad para abordar los síntomas cardinales de la enfermedad, aunque siguen existiendo casos resistentes al tratamiento antinarcoléptico. Los fármacos con mecanismos de acción sobre receptores del sistema histaminérgico pueden resultar de gran utilidad en estos casos.(AU)

Introduction: Narcolepsy type 1 is a focal degenerative disease of the hypothalamus that selectively affects orexin (hypocretin)-producing neurons. It presents multiple clinical manifestations, both in wakefulness and in sleep. The symptoms are often so disruptive that they cause enormous suffering and impair patients’ quality of life. Although a non-pharmacological approach is sometimes sufficient, the vast majority of patients need medication for adequate clinical management. Case report: A male who, at 43 years of age, began to present acutely with excessive daytime sleepiness and episodes of cataplexy. After a thorough examination, he was diagnosed with narcolepsy type 1. Throughout the course of the disease, he was prescribed antidepressants, neurostimulants and sodium oxybate, in monotherapy or in combination. The response to pharmacological treatment was insufficient and accompanied by numerous side effects. Following the introduction of pitolisant, there was a marked improvement in his symptoms and a reduction in the dose of the other drugs and their adverse effects was achieved. Conclusion: A number of measures are now available to address the cardinal symptoms of the disease, although there are still cases that are resistant to anti-narcoleptic treatment. Drugs with mechanisms of action that act upon receptors in the histaminergic system can be very useful in these cases.(AU)

Inhibition of biofilm formation, quorum sensing and virulence factor production in Pseudomonas aeruginosa PAO1 by selected LasR inhibitors / Inhibición de la formación de biopelículas, detección de quórum y producción de factores de virulencia en Pseudomonas aeruginosa PAO1 mediante inhibidores de LasR seleccionados

The quorum sensing network of Pseudomonas aeruginosa mediates the regulation of genes controlling biofilm formation and virulence factors. The rise of drug resistance to Pseudomonas aeruginosa infections has made quorum sensing–regulated biofilm formation in clinical settings a major issue. In the present study, LasR inhibitors identified in our previous study were evaluated for their antibiofilm and antiquorum sensing activities against P. aeruginosa PAO1. The compounds selected were (3-[2-(3,4-dimethoxyphenyl)-2-(1H-indol-3-yl)ethyl]-1-(2-fluorophenyl)urea) (C1), (3-(4-fluorophenyl)-2-[(3-methylquinoxalin-2-yl)methylsulfanyl]quinazolin-4-one) (C2) and (2-({4-[4-(2-methoxyphenyl)piperazin-1-yl]pyrimidin-2-yl}sulfanyl)-N-(2,4,6-trimethylphenyl)acetamide) (C3). The minimum inhibitory concentrations of C1 and C2 were 1000 μM, whereas that of C3 was 500 μM. At sub-MICs, the compounds showed potent antibiofilm activity without affecting the growth of P. aeruginosa PAO1. Electron microscopy confirmed the disruption of biofilm by the selected compounds. The antiquorum sensing activity of the compounds was revealed by the inhibition of violacein in Chromobacterium violaceum and the inhibition of swimming and swarming motilities in P. aeruginosa PAO1. Furthermore, the compounds also attenuated the production of quorum sensing–mediated virulence factors. The qRT-PCR revealed the downregulation of quorum sensing regulatory genes, namely lasI, lasR, rhlI, rhlR, lasB, pqsA and pqsR. The selected compounds also exhibited lower cytotoxicity against peripheral blood lymphocytes. Thus, this study could pave a way to explore these compounds for the development of therapeutic agent against Pseudomonas aeruginosa biofilm–related infections.(AU)

Virulence, antimicrobial resistance, and molecular characteristics of carbapenem-resistant Klebsiella pneumoniae in a hospital in Shijiazhuang City from China / Virulencia, resistencia a los antimicrobianos y características moleculares de Klebsiella pneumoniae resistente a carbapenémicos en un hospital de la ciudad de Shijiazhuang de China

Carbapenem-resistant Klebsiella pneumoniae (CRKP), as one of the most common drug-resistant bacteria threatening human health, is hyper-resistant to multiple antimicrobial drugs and carbapenems, which can be dealt with only limited clinical treatment options. This study described the epidemiological characteristics of CRKP in this tertiary care hospital from 2016 to 2020. Specimen sources included blood, sputum, alveolar lavage fluid, puncture fluid, secretions from a burn wound, and urine. Among the 87 carbapenem-resistant strains, ST11 was the predominant isolate, followed by ST15, ST273, ST340, and ST626. These STs were in broad agreement with the STs defined by pulsed-field gel electrophoresis clustering analysis in discriminating clusters of related strains. Most CRKP isolates contained the blaKPC-2 gene, some isolates carried the blaOXA-1, blaNDM-1, and blaNDM-5 genes, and the isolates carrying carbapenem resistance genes were more resistant to the antimicrobials of β-lactams, carbapenems, macrolides, and fluoroquinolone. The OmpK35 and OmpK37 genes were detected in all CRKP strains, and the Ompk36 gene was detected in some CRKP strains. All detected OmpK37 had 4 mutant sites, and OmpK36 had 11 mutant sites, while no mutant sites were found in OmpK35. More than half of the CRKP strains contained the OqxA and OqxB efflux pump genes. The virulence genes were most commonly combined with urea-wabG-fimH-entB-ybtS-uge-ycf. Only one CRKP isolate was detected with the K54 podoconjugate serotype. This study elucidated the clinical epidemiological features and molecular typing of CRKP, and grasped the distribution of drug-resistant genotypes, podocyte serotypes, and virulence genes of CRKP, providing some guidance for the subsequent treatment of CRKP infection.(AU)

Antimicrobial resistance and molecular epidemiology of carbapenem-resistant Escherichia coli from urinary tract infections in Shandong, China / Resistencia a los antimicrobianos y epidemiología molecular de Escherichia coli resistente a carbapenémicos procedente de infecciones del tracto urinario en Shandong, China

Objectives: Urinary tract infection (UTI) is one of the most common extraintestinal infections, and uropathogenic Escherichia coli (UPEC) is the main cause of UTIs. However, the ability to treat UTI has been compromised by the increase in antimicrobial resistance, especially carbapenem resistance. Here, we aimed to characterize the antimicrobial resistance and molecular epidemiology of carbapenem-resistant UPEC isolated in Shandong, China. Methods: In total, 17 carbapenem-resistant UPEC (CR-UPEC) isolates were collected from July 2017 to May 2020 in the Shandong Provincial Hospital. Whole-genome sequencing and bioinformatics analyses were performed to understand the molecular epidemiology of CR-UPEC. Phylogenetic groups, drug resistance genes, biofilm formation, and virulence-related gene profiles of the isolates were analyzed. Plasmid profiling and conjugation assay were performed to evaluate the ability to transfer carbapenem resistance-related genes to other E. coli isolates. Biofilm formation was also evaluated, as it is important for the persistence of infectious diseases. Results: We observed that 15 out of 17 CR-UPEC strains were blaNDM producers, among which 4 isolates could transfer blaNDM to recipient cells. The predominant sequence type was ST167 (6/17), followed by ST410 (3/17). The most prevalent phylogenetic group was phylogenetic group A (10/17), followed by phylogenetic group C (3/17). One isolate was resistant to polymyxin, which was caused by the carriage of a transferable plasmid harboring mcr-1. Statistical analysis did not reveal any significant difference in the carriage rate of fimbriae-coding genes between strong and weak biofilm producers. Conclusions: Our observations may assist in developing new therapeutic methods for drug-resistant organisms.(AU)

Development of diagnostic tests for pathogen identification and detection of antimicrobial resistance on WHO global priority pathogens using modular real-time nucleic acid amplification test / Desarrollo de pruebas de diagnóstico para la identificación de patógenos y la detección de resistencia a los antimicrobianos en patógenos prioritarios a nivel mundial de la OMS utilizando pruebas modulares de amplificación de ácidos nucleicos en tiempo real

Background: Concerns regarding antimicrobial resistance (AMR) have resulted in the World Health Organization (WHO) designating so-called global priority pathogens (GPPs). However, little discussion has focused on the diagnosis of GPPs. To enable the simultaneous identification of pathogens and AMR, we developed a modular real-time nucleic acid amplification test (MRT-NAAT). Methods: Sequence-specific primers for each modular unit for MRT-NAAT pathogen identification and AMR sets were designed. The composition of the reaction mixture and the real-time PCR program were unified irrespective of primer type so to give MRT-NAAT modularity. Standard strains and clinical isolates were used to evaluate the performance of MRT-NAAT by real-time PCR and melting curve analysis. Probit analysis for the MRT-NAAT pathogen identification set was used to assess the limit of detection (LoD). Results: The MRT-NAAT pathogen identification set was made up of 15 modular units 109–199 bp in product size and with a Tms of 75.5–87.5 °C. The LoD was < 15.548 fg/μL, and nine modular units successfully detected the target pathogens. The MRT-NAAT AMR set included 24 modular units 65–785 bp in product size with a Tms of 75.5–87.5 °C; it showed high performance for detecting GPP target genes and variants. Conclusions; MRT-NAAT enables pathogen identification and AMR gene detection and is time-effective. By unifying the reaction settings of each modular unit, the modularity where combinations of primers can be used according to need could be achieved. This would greatly help in reflecting the researcher’s need and the AMR status of a certain region while successfully detecting pathogens and AMR genes.(AU)

Prevalence and antimicrobial resistance of food safety related Vibrio species in inland saline water shrimp culture farms / Prevalencia y resistencia a los antimicrobianos de especies de Vibrio relacionadas con la inocuidad de los alimentos en granjas de cultivo de camarones de aguas saladas continentales

This study evaluated the potential pathogenicity and antimicrobial resistance (AMR) of Vibrio species isolated from inland saline shrimp culture farms. Out of 200 Vibrio isolates obtained from 166 shrimp/water samples, 105 isolates were identified as V. parahaemolyticus and 31 isolates were identified as V. alginolyticus and V. cholerae, respectively. During PCR screening of virulence-associated genes, the presence of the tlh gene was confirmed in 70 and 19 isolates of V. parahaemolyticus and V. alginolyticus, respectively. Besides, 10 isolates of V. parahaemolyticus were also found positive for trh gene. During antibiotic susceptibility testing (AST), very high resistance to cefotaxime (93.0%), amoxiclav (90.3%), ampicillin (88.2%), and ceftazidime (73.7%) was observed in all Vibrio species. Multiple antibiotic resistance (MAR) index values of Vibrio isolates ranged from 0.00 to 0.75, with 90.1% of isolates showing resistance to ≥ 3 antibiotics. The AST and MAR patterns did not significantly vary sample-wise or Vibrio species-wise. During the minimum inhibitory concentration (MIC) testing of various antibiotics against Vibrio isolates, the highest MIC values were recorded for amoxiclav followed by kanamycin. These results indicated that multi-drug resistant Vibrio species could act as the reservoirs of antibiotic resistance genes in the shrimp culture environment. The limited host range of 12 previously isolated V. parahaemolyticus phages against V. parahaemolyticus isolates from this study indicated that multiple strains of V. parahaemolyticus were prevalent in inland saline shrimp culture farms. The findings of the current study emphasize that routine monitoring of emerging aquaculture areas is critical for AMR pathogen risk assessment.(AU)

Molecular characterization of colistin resistance genes in a high-risk ST101/KPC-2 clone of Klebsiella pneumoniae in a University Hospital of Split, Croatia / Caracterización molecular de genes de resistencia a la colistina en un clon ST101/KPC-2 de alto riesgo de Klebsiella pneumoniae en un Hospital Universitario de Split, Croacia

Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-KP) has become a major concern worldwide due to multidrug resistance and the ability to spread locally and globally. Infections caused by KPC-KP are great challenge in the healthcare systems because these are associated with longer hospitalization and high mortality. The emergence of colistin resistance has significantly reduced already limited treatment options. This study describes the molecular background of colistin-resistant KPC-KP isolates in the largest hospital in southern Croatia. Thirty-four non-duplicate colistin-resistant KPC-KP isolates were collected during routine work from April 2019 to January 2020 and from February to May 2021. Antimicrobial susceptibility was determined using disk diffusion, broth microdilution, and the gradient strip method. Carbapenemase was detected with an immunochromatographic test. Identification of blaKPC and mcr genes or mutations in pmrA, pmrB, mgrB, phoP, and phoQ genes were performed by polymerase chain reaction (PCR) and positive products were sequenced. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used for epidemiological analysis. All isolates were multidrug-resistant, with colistin minimum inhibitory concentrations (MICs) from 4 to >16 mg/L, and all harbored blaKPC-2 and had a single point mutation in the mgrB gene resulting in a premature stop codon, with the exception of one isolate with four point mutations corresponding to stop codons. All isolates were negative for mcr genes. PFGE analysis identified a single genetic cluster, and MLST revealed that all isolates belonged to sequence type 101 (ST101). These results show emergence of the high-risk ST101/KPC-2 clone of K. pneumoniae in Croatia as well as appearance of colistin resistance due to mutations in the mgrB gene. Molecular analysis of epidemiology and possible resistance mechanisms are important to develop further strategies to combat such threats.(AU)

Regulation of ABC transporters by sex steroids may explain differences in drug resistance between sexes

Drug efficacy is dependent on the pharmacokinetics and pharmacodynamics of therapeutic agents. Tight junctions, detoxification enzymes, and drug transporters, due to their localization on epithelial barriers, modulate the absorption, distribution, and the elimination of a drug. The epithelial barriers which control the pharmacokinetic processes are sex steroid hormone targets, and in this way, sex hormones may also control the drug transport across these barriers. Thus, sex steroids contribute to sex differences in drug resistance and have a relevant impact on the sex-related efficacy of many therapeutic drugs. As a consequence, for the further development and optimization of therapeutic strategies, the sex of the individuals must be taken into consideration. Here, we gather and discuss the evidence about the regulation of ATP-binding cassette transporters by sex steroids, and we also describe the signaling pathways by which sex steroids modulate ATP-binding cassette transporters expression, with a focus in the most important ATP-binding cassette transporters involved in multidrug resistance. (AU)

Mycoplasma pneumoniae y resistencias a macrólidos: ¿Conocemos la situación en Europa? / Macrolide-resistant Mycoplasma pneumoniae: Do we know the situation in Europe?

Mycoplasma pneumoniae es una bacteria que carece de pared celular. Produce infecciones en todo el mundo, en brotes epidémicos cada 4-7 años, o en forma endémica. Sus manifestaciones clínicas se producen mayoritariamente en el tracto respiratorio y es una causa común de neumonía atípica. El tratamiento se realiza con macrólidos, tetraciclinas o fluoroquinolonas. Desde el año 2000 se ha detectado un aumento de resistencias a macrólidos a nivel mundial, siendo más frecuentes en Asia. En Europa la frecuencia de resistencias oscila entre el 1% y 25% en diferentes países. La combinación de técnicas moleculares y serológicas aporta una alta sensibilidad en la confirmación diagnóstica, siendo de gran utilidad para la detección y control de brotes de M. pneumoniae. La detección de resistencia a macrólidos requiere una técnica de secuenciación. (AU)

Mycoplasma pneumoniae is a bacterium that lacks a cell wall. It produces infections all It produces infections world-wide, in epidemic outbreaks every 4-7 years, or endemically. Its clinical manifestations occur mostly in the respiratory tract and it is a common cause of atypical pneumonia. The treatment is with macrolides, tetracyclines or fluoroquinolones. Since 2000, an increase in resistance to macrolides has been detected worldwide, being more frequent in Asia. In Europe the frequency of resistance ranges between 1% and 25%, depending on the country. Molecular techniques and serology techniques provides very high sensitivity in diagnostic confirmation, being very useful for detecting and controlling M. pneumoniae outbreaks. The detection of resistance to macrolides requires a sequencing technique.

Cambios en la resistencia antibiótica en episodios de bacteriospermia sintomática: Evolución en un área de salud del sudeste español / Antibiotic resistance changes in episodes of symptomatic bacteriospermia: development in a health area of southeast Spain

Objetivo: La prostatitis crónica bacteriana (PCB) es una entidad de difícil diagnóstico clínico y tratamiento, siendo el estudio microbiológico del semen la principal prueba diagnóstica. Este estudio tuvo como objetivo determinar la etiología y la resistencia antibiótica en pacientes con bacteriospermia sintomática (BPS) en nuestro medio. Material y métodos: Se ha realizado un estudio descriptivo, transversal y retrospectivo, en un Hospital Regional del sudeste español. Los participantes fueron pacientes asistidos en las consultas del Hospital con clínica compatible con PCB entre 2016 y 2021. Se recogieron y analizaron los resultados del estudio microbiológico de la muestra de semen. Se evaluó la etiología y la tasa de resistencia antibiótica de los episodios de BPS. Resultados: El principal microorganismo detectado es Enterococcus faecalis (34,89%), seguido por Ureaplasma spp. (13,74%) y Escherichia coli (10,98%). La tasa de resistencia antibiótica de E. faecalis a las quinolonas (11%) es inferior a estudios previos, mientras que, para E. coli ha sido superior (35%). Destaca la baja tasa de resistencia que muestran E. faecalis y E. coli a fosfomicina y nitrofurantoína. Conclusiones: En las BPS las bacterias grampositivas y las atípicas se establecen como los principales agentes causales de esta entidad. Esto obliga a replantear la estrategia terapéutica utilizada, lo cual evitará el aumento en las resistencias antibióticas, las recidivas y la cronicidad de esta patología. (AU)

Background: Chronic bacterial prostatitis (CBP) is an entity of difficult clinical diagnosis and treatment, being the microbiological study of semen the main diagnostic test. This study aimed to determine the etiology and antibiotic resistance in patients with symptomatic bacteriospermia (SBP) in our environment. Material and methods: A cross-sectional and retrospective descriptive study has been carried out from a Regional Hospital of the Spanish Southeast. The participants were patients assisted in the consultations of the Hospital with clinic compatible with CBP, between 2016 and 2021. The interventions were collection and analysis of the results derived from the microbiological study of the semen sample. The main determinations were the etiology and rate of antibiotic resistance of BPS episodes are analyzed. Results: The main isolated microorganism is Enterococcus faecalis (34.89%), followed by Ureaplasma spp. (13.74%) and Escherichia coli (10.98%). The rate of antibiotic resistance of E. faecalis to quinolones (11%) is lower than previous studies, while for E. coli it has been higher (35%). The low rate of resistance shown by E. faecalis and E. coli to fosfomycin and nitrofurantoin stands out. Conclusions: In the SBP, gram-positive and atypical bacteria are established as the main causative agents of this entity. This forces us to rethink the therapeutic strategy used, which will avoid the increase in antibiotic resistance, recurrences, and chronicity of this pathology.

Epidemiology, molecular characterization, and drug resistance of IncHI5 plasmids from Enterobacteriaceae / Epidemiología, caracterización molecular y farmacorresistencia de plásmidos IncHI5 de Enterobacteriaceae

The increasingly frequent occurence of IncHI5 plasmids has attracted worldwide attention. The aim of this study was to perform an in-depth bioinformatics analysis to determine the genetic characteristics and global distribution of all IncHI5 plasmids. The geographic distribution and epidemiology of all IncHI5 plasmids from GenBank were analyzed based on relevant literature reports and background information from the National Center for Biotechnology Information (NCBI). Detailed annotation of antibiotic resistance genes was performed. A total of 65 IncHI5 plasmid genomes were collected in GenBank. All IncHI5 plasmids were carried by Enterobacteriaceae, of which Klebsiella pneumoniae accounted for the largest proportion (50%, 33/65). The host bacterium of IncHI5 plasmids was mainly isolated from Homo Sapiens (81%, 53/65). All strains carrying IncHI5 plasmids were mainly distributed in China (83%, 54/65). Evolutionary analysis can divide IncHI5 plasmids into two groups, namely Groups I/II, of which Group II was more widely distributed worldwide. This study showed that Enterobacteriaceae, especially Klebsiella, was the main host for IncHI5 plasmid. Almost all IncHI5 plasmids carried multiple types of antibiotic resistance genes, related to Tn1696 or Tn6535. The IncHI5 plasmids should be of continuing interest as good repositories for antibiotic resistance genes.(AU)

Resistencia bacteriana, una crisis actual / Bacterial resistance, a current crisis

La resistencia bacteriana es una constante batalla que representa un problema de Salud Pública. Tan es así que la Organización Mundial de la Salud (OMS) la considera de sus prioridades en salud, debido al impacto que genera tanto en la salud (dado que proyecciones recientes indican que para 2050 se producirán más muertes por esta causa que las ocasionadas actualmente por elcáncer), como a su impacto económico (que, de acuerdo a un estudio reciente en el Reino Unido(1), costará a la economía mundialun estimado de 100 billones de dólares anualmente). La veloz aparición de bacterias multirresistentes y panresistentes es unfenómeno de índole mundial, cuestionando la eficacia antibiótica. Implementar protocolos y recomendaciones es vital, de igualforma que es necesario conciencias al personal sanitario, tomando como base el conocimiento de generación de resistencia y suimpacto a través de los años, potenciado por la actual pandemia de la COVID-19.(AU)

Bacterial resistance is a constant battle representing a Public Health trouble. So much, that the World Health Organization consi-derate Public Health as a priority in health, due to the impact that generates as much as in health (giving that recent projectionsindicate that by 2050 it’ll be produced more deaths because of this than the ones occasioned because of cancer) as its economicimpact (which, according to a recent study in the United Kingdom(1), it’ll cost the world’s economy an estimated of 100 trilliondollars). The quick appearance of multidrug-resistant and pandrug-resistant bacteria is a world nature phenomenon, questioningthe antibiotics efficiency. Implement protocols and recommendations is essential, just as essential and necessary as give aware-ness to health personnel, taking as base the knowledge of resistance generation and its impact through the years, empoweredby the actual pandemic of COVID 19.(AU)

Antimicrobial resistance in Gram-negative bacilli in Spain: an experts’ view / Resistencia a antimicrobianos en bacilos gramnegativos en España: una visión de expertos

Background: Antibiotic resistance in Gram-negative bacilli poses a serious problem for public health. In hospitals, in addition to high mortality rates, the emergence and spread of resistance to practically all antibiotics restricts therapeutic options against serious and frequent infections. Objectives: The aim of this work is to present the views of a group of experts on the following aspects regarding resistance to antimicrobial agents in Gram-negative bacilli: 1) the current epidemiology in Spain, 2) how it is related to local clinical practice and 3) new therapies in this area, based on currently available evidence. Methodology: After reviewing the most noteworthy evidence, the most relevant data on these three aspects were presented at a national meeting to 99 experts in infectious diseases, clinical microbiology, internal medicine, intensive care medicine, anaesthesiology and hospital pharmacy. Results and conclusions: Subsequent local debates among these experts led to conclusions in this matter, including the opinion that the approval of new antibiotics makes it necessary to train the specialists involved in order to optimise how they use them and improve health outcomes; microbiology laboratories in hospitals must be available throughout a continuous timetable; all antibiotics must be available when needed and it is necessary to learn to use them correctly; and the Antimicrobial Stewardship Programs (ASP) play a key role in quickly allocating the new antibiotics within the guidelines and ensure appropriate use of them. (AU)

Contexto: La resistencia a los antibióticos en bacilos gramnegativos representa un grave problema de salud pública. En el hospital, además de unas elevadas tasas de mortalidad, la aparición y propagación de resistencias a la práctica totalidad de los antibióticos limita las opciones terapéuticas frente a infecciones graves y frecuentes. Objetivos: Este trabajo tiene por objetivo dar a conocer la visión de un grupo de expertos en los siguientes aspectos respecto a la resistencia a agentes antimicrobianos en bacilos gramnegativos: 1) la epidemiología actual en España, 2) su relación con la práctica clínica local y 3) las novedades terapéuticas en este ámbito, fundamentada en la evidencia actualmente disponible. Metodología: Tras la revisión de la evidencia más destacada, los datos más relevantes de estos 3 aspectos fueron presentados en una reunión nacional ante 99 expertos en enfermedades infecciosas, microbiología clínica, medicina interna, medicina intensiva, anestesiología y farmacia hospitalaria. Resultados y conclusiones: De debates locales posteriores entre estos expertos se extrajeron conclusiones al respecto entre las que se destacan que la aprobación de nuevos antibióticos hace necesaria la formación de los especialistas implicados para optimizar su uso y mejorar los resultados en salud; los laboratorios de Microbiología de los hospitales deben estar disponibles en horario continuado; todos los antibióticos deben estar disponibles para cuando sean necesarios y se debe aprender a usarlos de forma correcta; y los Programas de Optimización del Uso de Antimicrobianos (PROA) desempeñan una labor clave en ubicar de forma ágil los nuevos antibióticos en las guías y asegurar un uso apropiado de los mismos. (AU)

Reconstrucción ancestral de una B-lactamasa y comparativa con sus homólogos actuales / Ancestral reconstruction of a B-lactamase and comparison with its extant proteins

Las B-lactamasas son proteínas de origen bacteriano que se caracterizan por hidrolizar los antibióticos B-lactámicos, confiriendo resistencia microbiana ante estos. Son una familia heterogénea de proteínas muy relevantes desde el punto de vista sanitario debido a la facilidad que presentan para adquirir resistencia a nuevos fármacos por su alta capacidad de evolución.La evolución in vitro de estas proteínas ha servido, no solo para desarrollar su caracterización y mejorar su conocimiento, sino como una nueva línea de investigación que permite identificar de manera predictiva residuos implicados en la adquisición de resistencia frente antibióticos. Al mismo tiempo, el método de reconstrucción ancestral de proteínas se ha revelado como una herramienta novedosa y útil para comprender la evolución de las β- lactamasas y entender algunas de sus características como es su promiscuidad. En este trabajo, se ha realizado un estudio de B-lactamasas ancestrales reconstruidas a partir de la filogenia de B-lactamasas existentes de clase A. De las cuatro proteínas ancestrales estudiadas, se ha obtenido una que es funcional y se ha comparado su actividad hidrolítica con la de cuatro de sus homólogos actuales frente a ocho fármacos B-lactámicos. Se ha comprobado que esta proteína ancestral tiene una actividad frente aantibióticos más generalista que cualquier de las proteínas actuales estudiadas. Además, la proteína ancestral activa mostró más resistencia frente a uno de los fármacos utilizados que el resto de B-lactamasas existentes. Finalmente se han discutido estos resultados y a partir de ellos se argumenta por qué las secuencias ancestrales reconstruidas pueden ser un punto de partida muy atractivo a la hora de realizar evolución dirigida de proteínas para la obtención de proteínas de interés biotecnológico. (AU)

The B-lactamases are proteins of bacterial origin that are characterized by hydrolyzing antibiotics B-lactams, conferring microbial resistance against them. They are a heterogeneous family of proteins very relevant from a health point of view due to the ease they present to acquire resistance to new drugs due to their high capacity for evolution.The in vitro evolution of these proteins has served not only to develop their characterization and improve their knowledge, but as a new line of research that allows to predictively identify residues involved in the acquisition of antibiotic resistance.At the same time, the method of ancestral protein reconstruction has been revealed as a novel and useful tool to understand the evolution of B-lactamases and understand some of their characteristics such as their promiscuity.In this work, a study of ancestral B-lactamases reconstructed from the phylogeny of existing class A B-lactamases has been carried out. Of the four ancestral proteins studied, one has been obtained that is functional and has compared its hydrolytic activity with that of four of its current counterparts against eight β-lactam drugs. This ancestral protein has been shown to have a more generalistic antibiotic activity than any of the current proteins studied. In addition, the active ancestral protein showed more resistance to one of the drugs used than the rest of B-lactamases existing. Finally these results have been discussed and from them it is argued why reconstructed ancestral sequences can be a very attractive starting point when it comes to direct evolution of proteins for obtaining proteins of biotechnological interest.