Epidemiology, resistance genomics and susceptibility of Acinetobacter species: results from the 2020 Spanish nationwide surveillance study.

BackgroundAs increasing antibiotic resistance in Acinetobacter baumannii poses a global healthcare challenge, understanding its evolution is crucial for effective control strategies.AimWe aimed to evaluate the epidemiology, antimicrobial susceptibility and main resistance mechanisms of Acinetobacter spp. in Spain in 2020, and to explore temporal trends of A. baumannii.MethodsWe collected 199 single-patient Acinetobacter spp. clinical isolates in 2020 from 18 Spanish tertiary hospitals. Minimum inhibitory concentrations (MICs) for nine antimicrobials were determined. Short-read sequencing was performed for all isolates, and targeted long-read sequencing for A. baumannii. Resistance mechanisms, phylogenetics and clonality were assessed. Findings on resistance rates and infection types were compared with data from 2000 and 2010.ResultsCefiderocol and colistin exhibited the highest activity against A. baumannii, although colistin susceptibility has significantly declined over 2 decades. A. non-baumannii strains were highly susceptible to most tested antibiotics. Of the A. baumannii isolates, 47.5% (56/118) were multidrug-resistant (MDR). Phylogeny and clonal relationship analysis of A. baumannii revealed five prevalent international clones, notably IC2 (ST2, n = 52; ST745, n = 4) and IC1 (ST1, n = 14), and some episodes of clonal dissemination. Genes bla OXA-23, bla OXA-58 and bla OXA-24/40 were identified in 49 (41.5%), eight (6.8%) and one (0.8%) A. baumannii isolates, respectively. ISAba1 was found upstream of the gene (a bla OXA-51-like) in 10 isolates.ConclusionsThe emergence of OXA-23-producing ST1 and ST2, the predominant MDR lineages, shows a pivotal shift in carbapenem-resistant A. baumannii (CRAB) epidemiology in Spain. Coupled with increased colistin resistance, these changes underscore notable alterations in regional antimicrobial resistance dynamics.

Activity of cefiderocol and innovative ß-lactam/ß-lactamase inhibitor combinations against isogenic strains of Escherichia coli expressing single and double ß-lactamases under high and low permeability conditions.

OBJECTIVES: To analyse the impact of the most clinically relevant ß-lactamases and their interplay with low outer membrane permeability on the activity of cefiderocol, ceftazidime/avibactam, aztreonam/avibactam, cefepime/enmetazobactam, cefepime/taniborbactam, cefepime/zidebactam, imipenem/relebactam, meropenem/vaborbactam, meropenem/xeruborbactam and meropenem/nacubactam against recombinant Escherichia coli strains. METHODS: We constructed 82 E. coli laboratory transformants expressing the main ß-lactamases circulating in Enterobacterales (70 expressing single ß-lactamase and 12 producing double carbapenemase) under high (E. coli TG1) and low (E. coli HB4) permeability conditions. Antimicrobial susceptibility testing was determined by reference broth microdilution. RESULTS: Aztreonam/avibactam, cefepime/zidebactam, cefiderocol, meropenem/xeruborbactam and meropenem/nacubactam were active against all E. coli TG1 transformants. Imipenem/relebactam, meropenem/vaborbactam, cefepime/taniborbactam and cefepime/enmetazobactam were also highly active, but unstable against most of MBL-producing transformants. Combination of ß-lactamases with porin deficiency (E. coli HB4) did not significantly affect the activity of aztreonam/avibactam, cefepime/zidebactam, cefiderocol or meropenem/nacubactam, but limited the effectiveness of the rest of carbapenem- and cefepime-based combinations. Double-carbapenemase production resulted in the loss of activity of most of the compounds tested, an effect particularly evident for those E. coli HB4 transformants in which MBLs were present. CONCLUSIONS: Our findings highlight the promising activity that cefiderocol and new ß-lactam/ß-lactamase inhibitors have against recombinant E. coli strains expressing widespread ß-lactamases, including when these are combined with low permeability or other enzymes. Aztreonam/avibactam, cefiderocol, cefepime/zidebactam and meropenem/nacubactam will help to mitigate to some extent the urgency of new compounds able to resist MBL action, although NDM enzymes represent a growing challenge against which drug development efforts are still needed.

Assessment of the Degree of Clinical Suspicion of 21-Hydroxylase Deficiency Prior to the Newborn Screening Result.

The aim of the study was to analyze the clinical suspicion and where patients were when they received the positive result of the neonatal screening for CAH 21OHD. The present data derived from a retrospective analysis of a relatively large group of patients with classical CAH 21OHD patients nosed by newborn screening in Madrid, Spain. During the period from 1990 to 2015 of this study 46 children were diagnosed with classical 21OHD [36 with the salt-wasting (SW) form and 10 with simple virilizing (SV)]. In 38 patients, the disease had not been suspected before the neonatal screening result (30 SW and 8 SV). Thirty patients (79%) were at home without suspicion of any disease, as healthy children, 3 patients (8%) were at home pending completion of the study due to clinical suspicion of any disease (ambiguous genitalia, cryptorchidism) and 5 patients (13%) were admitted to the hospital for reasons unrelated to CAH (sepsis, jaundice, hypoglycemia). It is relevant to note that 69.4% of patients (25/36) with SW form were at home with potential risk of adrenal crisis. Six females had been incorrectly labeled as male at birth. The most frequent reason for clinical suspicion was genital ambiguity in women followed by family history of the disease. Neonatal screening provided better results than clinical suspicion. In the majority of patients with 21OHD the diagnosis by screening was anticipated to the clinical suspicion of the disease even in female patients with ambiguous genitalia.

Interplay between OXA-10 ß-Lactamase Production and Low Outer-Membrane Permeability in Carbapenem Resistance in Enterobacterales.

The OXA-10 class D ß-lactamase has been reported to contribute to carbapenem resistance in non-fermenting Gram-negative bacilli; however, its contribution to carbapenem resistance in Enterobacterales is unknown. In this work, minimum inhibitory concentrations (MICs), whole genome sequencing (WGS), cloning experiments, kinetic assays, molecular modelling studies, and biochemical assays for carbapenemase detection were performed to determine the impact of OXA-10 production on carbapenem resistance in two XDR clinical isolates of Escherichia coli with the carbapenem resistance phenotype (ertapenem resistance). WGS identified the two clinical isolates as belonging to ST57 in close genomic proximity to each other. Additionally, the presence of the blaOXA-10 gene was identified in both isolates, as well as relevant mutations in the genes coding for the OmpC and OmpF porins. Cloning of blaOXA-10 in an E. coli HB4 (OmpC and OmpF-deficient) demonstrated the important contribution of OXA-10 to increased carbapenem MICs when associated with porin deficiency. Kinetic analysis showed that OXA-10 has low carbapenem-hydrolysing activity, but molecular models revealed interactions of this ß-lactamase with the carbapenems. OXA-10 was not detected with biochemical tests used in clinical laboratories. In conclusion, the ß-lactamase OXA-10 limits the activity of carbapenems in Enterobacterales when combined with low permeability and should be monitored in the future.

[Impact of the COVID-19 pandemic on Neonatal Screening for Congenital Hypothyroidism.] / Impacto de la pandemia por COVID-19 en el cribado neonatal de hipotirodismo congénito.

OBJECTIVE: The purpose of this paper was to describe the diagnosis, treatment and follow-up of patients diagnosed with congenital hypothyroidism (CH) by the Neonatal Screening Program in the Autonomous Community of Madrid during the state of alarm due to the COVID-19 health crisis. METHODS: The data were extracted from the retrospective analysis of patients diagnosed with CH and treated at the Clinical Diagnosis and Follow-up Center of CH located in the Pediatric Endocrinology Unit of the General University Hospital Gregorio Marañon. RESULTS: During the period between March 14 and June 21, 2020, 7 neonates were diagnosed with congenital hypothyroidism. The Screening Center contacted the Clinical Diagnosis and Follow-up Center urgently, with the location and clinical assessment of the patient on the same day, performing the usual complementary examinations in all of them according to clinical pathway. The median age of diagnosis was 15.5 days (range 7.00-24.00). The subsequent clinical and analytical follow-up was carried out in all cases according to the recommended times. All patients presented normalization of the thyroid function after two weeks of treatment. CONCLUSIONS: All patients seen at the Congenital Hypothyroidism Clinical Diagnosis and Follow-up Center during the alarm state period were diagnosed, treated and reevaluated following the usual clinical pathways without incidents. The current epidemiological situation of the COVID-19 pandemic has revealed the correct functioning of the circuit of the Congenital Hypothyroidism Screening Program in less favorable circumstances.

OBJETIVO: El objetivo de este trabajo fue mostrar el diagnóstico, tratamiento y seguimiento de los pacientes diagnosticados de hipotiroidismo congénito (HC) mediante el Programa de Cribado Neonatal en la Comunidad Autó-noma de Madrid durante el estado de alarma debido a la crisis sanitaria por la COVID-19. METODOS: Los datos fueron extraídos del análisis retrospectivo de pacientes diagnosticados de HC en el Centro de Diagnóstico y Seguimiento Clínico de HC, ubicado en la Unidad de Endocrinología Pediátrica del Hospital General Universitario Gregorio Marañón. RESULTADOS: Durante el período comprendido entre el 14 de marzo y el 21 de junio de 2020, siete neonatos fueron diagnosticados de HC. Desde el Centro de Cribado se contactó de forma urgente con el Centro Clínico de Diagnóstico y Seguimiento, con localización y valoración clínica del paciente el mismo día, realizándose las exploraciones complementarias habituales en todos ellos según la vía clínica. La edad mediana del diagnóstico fue de 15,5 días (rango 7,00-24,00). El seguimiento clínico y analítico posterior se realizó en todos los casos acorde a los tiempos recomendados. Todos los pacientes presentaron normalización de la función tiroidea a las dos semanas de tratamiento. CONCLUSIONES: Todos los pacientes atendidos en el Centro Clínico de Diagnóstico y Seguimiento de Hipotiroidismo Congénito durante el período de estado de alarma son diagnosticados, tratados y reevaluados siguiendo la vía clínica habitual, sin incidencias. La situación epidemiológica actual de la pandemia por la COVID-19 pone de manifiesto el correcto funcionamiento del circuito del Programa de Cribado de Hipotiroidismo Congénito en circunstancias menos favorables.

[Analysis of the degree of clinical suspect in patients with congenital adrenal hyperplasia by 21-hydroxylase deficiency before obtaining the result of the newborn screening program of the autonomous Community of Madrid.] / Análisis del grado de sospecha clínica en pacientes con hiperplasia suprarrenal congénita por déficit de 21 hidroxilasa antes de la obtención del resultado del cribado neonatal en la Comunidad de Madrid.

OBJECTIVE: The objective of this study was to analyze the clinical suspicion and where the patients were when they received the result of the neonatal screening for 21 hydroxylase deficiency (21OHD). METHODS: The present data were derived from a retrospective analysis of a group of patients with classical 21OHD discovered by newborn screening and treated at the Center for Clinical Follow-up of the Autonomous Community of Madrid. Stadistic analysis of the data was performed using version 15.5 of the SPSS® software. RESULTS: During the period from 1990 to 2015 of this study 46 children were diagnosed with classical 21OHD [36 with the salt-wasting (SW) and 10 with simple virilizing form (SV)]. The median age at diagnosis for the patients with the SW and SV form were 8.0 (6.0-9.0) and 18.0 (14.5-37.5) days respectively (P=0.001). In 35 (76.1%) patients the disease had not been suspected before the result of newborn screening, 28 patients affected by SW form, with a potential risk of death due to adrenal crisis (of which, in addition 6 women with incorrect assignment of sex at birth) and 7 patients affected with SV form. Two thirds of the patients with classic forms identified by neonatal screening were in their homes without suspicion of any disease or pending any additional study. CONCLUSIONS: Neonatal screening provided better performance than clinical suspicion. In the majority of patients with 21OHD detected by newborn screening, the diagnosis by screening was anticipated to the clinical suspicion of the disease even in female patients with ambiguous genitalia.

OBJETIVO: El objetivo de este estudio fue analizar el grado de sospecha clínica y donde estaban los pacientes cuando recibieron el resultado del cribado neonatal por déficit de 21 hidroxilasa (21OHD). METODOS: Los datos presentados fueron extraídos del análisis retrospectivo de pacientes diagnosticados de formas clásicas de 21OHD mediante Programa de Cribado Neonatal y atendidos en el Centro de Seguimiento Clínico de la Comunidad Autónoma de Madrid. El análisis estadístico de los datos se realizó empleando la versión 15.5 del software SPSS®. RESULTADOS: Durante el período comprendido entre 1990 a 2015, 46 niños fueron diagnosticados de formas clásicas por 21OHD [36 con pérdida salina (PS) y 10 con forma virilizante simple (VS)]. La edad mediana al diagnóstico de los pacientes con forma PS y forma VS fue 8,0 (6,0-9,0) y 18,0 (14,5-37,5) días respectivamente (P=0,001). En 35 (76,1%) pacientes la enfermedad no había sido sospechada antes del resultado del cribado neonatal, 28 pacientes estaban afectados de forma PS, con potencial riesgo de muerte debido a crisis adrenal (de ellos, 6 eran además mujeres en las que se había realizado una asignación incorrecta de sexo al nacimiento) y 7 pacientes afectados de forma VS. Dos tercios de los pacientes con formas clásicas identificados por cribado neonatal estaban en sus domicilios sin sospecha de ninguna enfermedad ni pendientes de completar estudios. CONCLUSIONES: El cribado neonatal proporciona mejor rendimiento que la sospecha clínica. En la mayoría de pacientes con 21OHD detectados por cribado neonatal, el diagnostico por cribado fue previo a la sospecha clínica de la enfermedad incluso en pacientes mujeres con ambigüedad genital.