Detection of blaCTX-M-15 in an integrative and conjugative element in four extensively drug-resistant Haemophilus parainfluenzae strains causing urethritis.

Haemophilus parainfluenzae is a commensal organism with rising numbers of multidrug-resistant (MDR) strains. This pathogen is of increasing clinical relevance in urogenital infection. The aim of this work was to identify and characterise the molecular mechanisms of resistance associated with four cephalosporin-resistant H. parainfluenzae strains collected from patients with urethritis. Antimicrobial resistance was determined by microdilution following European Committee on Antimicrobial Susceptibility Testing criteria. Strains were then analysed by whole-genome sequencing to determine clonal relationship and the molecular basis of antimicrobial resistance. Finally, a phylogenetic analysis was performed on all urogenital MDR strains of H. parainfluenzae previously isolated in our hospital. All strains were resistant to ß-lactams, macrolides, tetracycline, fluoroquinolones, chloramphenicol, cotrimoxazole, and aminoglycosides. The resistance profile was compatible with the presence of an extended-spectrum ß-lactamase (ESBL). Whole-genome sequencing detected blaCTX-M-15 that conferred high minimum inhibitory concentrations to cephalosporins in two novel integrative and conjugative elements (ICEHpaHUB6 and ICEHpaHUB7) that also harboured a blaTEM-1 ß-lactamase. This study shows a novel blaCTX-M-15 ESBL carried in an integrative conjugative element in four extensively drug-resistant H. parainfluenzae strains. This resistance determinant could be transmitted to other sexually transmitted pathogens and this is a cause for concern.

Analysis of patient outcomes after urological surgery during the second and third waves of SARS-CoV-2 pandemic in a high incidence area.

OBJECTIVE: To analyze surgical safety through postoperative COVID-19 incidence and mortality at the urology department of a tertiary hospital located in Madrid (Spain). METHODS: Observational, prospective study including all patients undergoing urological surgery from 1st March 2020 to 28th February 2021. According to the hospital organization and local epidemiological situation we delimitate three epidemic waves. A set of screening and protective measures was applied from 4th May onwards. Demographic, baseline, surgical and perioperative variables, as well as postoperative outcomes, were collected. Telephone follow-up was performed at least 3 weeks after hospital discharge. RESULTS: 940 urological surgeries were performed, 12 of them had to be rescheduled due to active or recent SARS-CoV-2 infection identified by the screening protocol. Thirty-one patients developed COVID-19 (3.3% incidence) and 7 died (22.6% mortality). The average time to onset of symptoms was 62.6 days after discharge, being 25 cases attributable to community transmission. The remaining 6 cases, due to in-hospital transmission, had worse outcomes. Five of them were identified during the first wave, especially when no preoperative PCR was obtained. In contrast, during the second and third waves, fewer and milder cases were diagnosed, with just 1 in-hospital transmission among 857 urological patients. CONCLUSIONS: After implementing complete protective measures, postoperative in-hospital COVID-19 cases almost disappeared, even during the second and third waves. Most of the cases were due to community transmission and thus driven by the general epidemiological situation. While hospitals follow recommendations to avoid COVID-19 infection, urological surgery remains safe and can be maintained.

Promoting oral health among 6-year old children: The impact of social environment and feeding behavior.

OBJECTIVE: To determine the prevalence of dental caries and identify key associated factors, including eating habits, body mass index, and the presence of dental plaque in immigrant and indigenous children. BASIC RESEARCH DESIGN: Observational study with immigrant and native child populations (aged 6 years) with logistic regression analysis. SETTING: Oral Public Health Service in the Asturias region of Spain. PARTICIPANTS: The presence of dental caries (World Health Organization) and oral hygiene (O'Leary index) were determined in 166 children. Information on eating habits was collected through a survey on food consumption (European Food Safety Authority). RESULT: Children from immigrant families had more decayed and filled teeth index (2.59 vs. 0.21; Crude OR: 7.99 [95% CI 3.620-17.636]). In univariate analysis, protective factors were daily intake of milk and fruits (p ⟨0.001). In multivariate analysis, the presence of caries was associated with immigrant status, consumption of sugary foods, and being overweight, (OR: 13.179 [4.668-37.208]). CONCLUSION: Public Institutions must take into account our results to help reduce caries in immigrant children.

Evaluation of health care-associated infections following radical cystectomy. / Estudio de las infecciones relacionadas con la asistencia sanitaria tras la realización de cistectomía radical.

INTRODUCTION AND OBJECTIVE: Radical cystectomy is a complex surgery with a high rate of complications including infections, which lead to increased morbidity and mortality, longer hospital stay and higher costs. The aim of this work is to evaluate health care-associated infections (HAIs) in these patients, as well as associated microorganisms, antibiotic resistance profiles and risk factors. MATERIAL AND METHODS: Prospective study from 2012 to 2017. Epidemiologic variables, comorbidities and surgical variables are collected. The microorganisms involved and antibiotic susceptibility patterns are analyzed. RESULTS: 122 patients. Mean age 67 (SD:18,42). Mean hospital stay 23.5 days (18.42). HAIs rate of 45%, with predominant urinary tract infections (43%) and surgical wound infections (31%). Positive cultures in 78.6% of cases. Increased isolation of Enterococcus (18%) and Escherichia coli (13%). Forty-three percent of microorganisms were resistant to amoxicillin/ampicillin, 23% to beta-lactamases and 36% to quinolones. Empirical treatment was adequate in 87.5%. Hospital stay is increased (17 days, p< 0.05) due to HAIs. Lower rate of infectious complications in the laparoscopic vs. open approach (p< 0.001) and in orthotopic vs. ileal conduit diversion (p = 0.04) CONCLUSIONS: We found a high rate of HAIs in our radical cystectomy series, with predominant urinary tract and surgical wound infections. E.coli and Enterococcus spp. are the most frequently isolated microorganisms, with high rates of resistance to some commonly used antibiotics.

Urological surgery during SARS-CoV-2 pandemic. Descriptive analysis of the experience in a Urology Department across the pandemic phases. / Cirugía urológica durante la pandemia por SARS-CoV-2. Análisis descriptivo de la experiencia en un Servicio de Urología durante distintas fases epidemiológicas.

INTRODUCTION: The SARS-CoV-2 pandemic has changed the urological practice around the world. Our objective is to describe the outcomes presented by patients undergoing surgery in the urology department of a tertiary hospital, across the pandemic phases. METHODS: Observational, cohort study including all patients undergoing surgery from March 1 to May 14. According to the hospital organization, we identified three periods: there were no changes during the first two weeks (1st. period), the following seven weeks, when only urgent interventions were carried out after performance of nasopharyngeal swab test (2nd. period), and finally, elective surgery was resumed on May 4, after the implementation of a multidisciplinary screening protocol (3rd. period). Demographic, baseline, surgical and perioperative variables, as well as postoperative outcomes, were obtained in a retrospective (periods 1 and 2) and prospective (period 3) manner. Telephone follow-up was initiated at least 3 weeks after hospital discharge. RESULTS: 103 urological surgeries were performed, and 11 patients were diagnosed with COVID-19, 8 of them within the 1st. PERIOD: The diagnosis was already known in 1 patient, while the other 10 developed the disease in an average of 25 days after the intervention and 16,6 days after discharge. Of seven transplant patients, four got the infection. Three deaths were recorded due to the disease: a 69-year-old woman transplanted and two men over 80 with comorbidities and high anesthetic risk who underwent drainage of retroperitoneal abscess and retrograde intrarenal surgery, respectively. CONCLUSIONS: SARS-CoV-2 infection mainly affected renal transplant recipients or elderly patients with high anesthetic risk, during the first 2 weeks of the pandemic. After implementing preoperative PCR tests and a comprehensive screening protocol, cases were substantially reduced, and safe surgical procedures were achieved.

Design of an assistance protocol for the restart of scheduled urologic surgery in a COVID-19 epidemic period. / Diseño de un protocolo asistencial para el reinicio de la cirugía urológica programada en periodo de epidemia COVID-19.

OBJECTIVE: Design a care protocol to restart scheduled surgical activity in a Urology service of a third level hospital in the Community of Madrid, in a safe way for our patients and professionals in the context of the SARS-CoV-2 coronavirus epidemic. MATERIAL AND METHODS: A multidisciplinary group reviewed the different recommendations of the literature, national and international health organizations and scientific societies, as well as their application to our environment. Once scheduled surgery has restarted, the patients undergoing surgery for complications related to COVID-19 are being followed up. RESULTS: Since the resumption of surgical activity, 19 patients have been scheduled, of which 2 have been suspended for presenting COVID-19, one diagnosed by positive PCR for SARS-CoV-2, and another by laboratory and imaging findings compatible with this infection. With a median follow-up of 10 days (4-14 days), no complications related to COVID-19 were detected. CONCLUSIONS: Preliminary results indicate that the protocol designed to ensure the correct application of preventive measures against the transmission of coronavirus infection is being safe and effective.

Assessment of trimethoprim-sulfamethoxazole susceptibility testing methods for fastidious Haemophilus spp.

OBJECTIVES: To compare the determinants of trimethoprim-sulfamethoxazole resistance with established susceptibility values for fastidious Haemophilus spp., to provide recommendations for optimal trimethoprim-sulfamethoxazole measurement. METHODS: We collected 50 strains each of Haemophilus influenzae and Haemophilus parainfluenzae at Bellvitge University Hospital. Trimethoprim-sulfamethoxazole susceptibility was tested by microdilution, E-test and disc diffusion using both Mueller-Hinton fastidious (MH-F) medium and Haemophilus test medium (HTM) following EUCAST and CLSI criteria, respectively. Mutations in folA, folP and additional determinants of resistance were identified in whole-genome-sequenced isolates. RESULTS: Strains presented generally higher rates of trimethoprim-sulfamethoxazole resistance when grown on HTM than on MH-F, independent of the methodology used (average MIC 2.6-fold higher in H. influenzae and 1.2-fold higher in H. parainfluenzae). The main resistance-related determinants were as follows: I95L and F154S/V in folA; 3- and 15-bp insertions and substitutions in folP; acquisition of sul genes; and FolA overproduction potentially linked to mutations in -35 and -10 promoter motifs. Of note, 2 of 19 H. influenzae strains (10.5%) and 9 of 33 H. parainfluenzae strains (27.3%) with mutations and assigned as resistant by microdilution were inaccurately considered susceptible by disc diffusion. This misinterpretation was resolved by raising the clinical resistance breakpoint of the EUCAST guidelines to ≤30 mm. CONCLUSIONS: Given the routine use of disc diffusion, a significant number of strains could potentially be miscategorized as susceptible to trimethoprim-sulfamethoxazole despite having resistance-related mutations. A simple modification to the current clinical resistance breakpoint given by the EUCAST guideline for MH-F ensures correct interpretation and correlation with the reference standard method of microdilution.

Emerging non-13-valent pneumococcal conjugate vaccine (PCV13) serotypes causing adult invasive pneumococcal disease in the late-PCV13 period in Spain.

OBJECTIVE: An early reduction of adult invasive pneumococcal disease (IPD) was observed after the 13-valent pneumococcal conjugate vaccine (PCV13) introduction for children in Spain. We analysed the epidemiology of adult IPD in the late-PCV13 period. METHODS: This was a prospective multicentre study of adult IPD involving six hospitals. Strains were serotyped, genotyped and studied for antimicrobial susceptibility. The late-PCV13 period was compared with the pre- and early-PCV13 periods. RESULTS: A total of 2197 episodes were collected-949 in 2008-2009, 609 in 2012-2013 and 639 in 2015-2016. The initial decrease of IPD observed (from 12.3/100 000 to 8.1/100 000; 2008-2009 versus 2012-2013) plateaued in 2015-2016 (8.3/100 000). IPD due to PCV13 serotypes decreased (from 7.7 to 3.5 to 2.3/100 000; p < 0.05), whereas IPD caused by non-PCV13 serotypes increased (from 4.5 to 4.6 to 6.0/100 000; p < 0.05). The most frequent serotypes in the late-PCV13 period were: 8 (15.1%), 3 (10.5%), 12F (7.9%) and 9N (5.4%). These serotypes were related to major genotypes: CC53 (59.8%) and CC404 (30.4%) for serotype 8, CC180 (64.1%) and CC260 (28.1%) for serotype 3, CC989 (91.7%) for serotype 12F and CC67 (84.8%) for serotype 9N. Penicillin-non-susceptibility (21.2%) was associated with serotypes 11A (CC156), 14 (CC156) and 19A (CC320), and macrolide-resistance was related to serotypes 24F and 19A. Rates of pneumococcal meningitis remained stable throughout the periods (ranges 0.9, 0.8 and 1.0/100 000). CONCLUSIONS: The initial decrease of adult IPD observed after PCV13 introduction for children has been balanced by the rise of non-PCV13 serotypes. The spread of antibiotic-resistant lineages related to non-PCV13 serotypes (11A and 24F) could be a threat for the treatment of serious pneumococcal diseases.

Insights into the population structure and pan-genome of Haemophilus influenzae.

The human-restricted bacterium Haemophilus influenzae is responsible for respiratory infections in both children and adults. While colonization begins in the upper airways, it can spread throughout the respiratory tract potentially leading to invasive infections. Although the spread of H. influenzae serotype b (Hib) has been prevented by vaccination, the emergence of infections by other serotypes as well as by non-typeable isolates (NTHi) have been observed, prompting the need for novel prevention strategies. Here, we aimed to study the population structure of H. influenzae and to get some insights into its pan-genome. We studied 305H. influenzae strains, enrolling 217 publicly available genomes, as well as 88 newly sequenced H. influenzae invasive strains isolated in Portugal, spanning a 24-year period. NTHi isolates presented a core-SNP-based genetic diversity about 10-fold higher than the one observed for Hib. The analysis of key factors involved in pathogenesis, such as lipooligosaccharides, hemagglutinating pili and High Molecular Weight-adhesins, suggests that NTHi shape its virulence repertoire, either by acquisition and loss of genes or by SNP-based diversification, likely towards host immune evasion and persistence. Discreet NTHi subpopulations structures are proposed based on core-genome supported with 17 candidate genetic markers identified in the accessory genome. Additionally, this study provides two bioinformatics tools for in silico rapid identification of H. influenzae serotypes and NTHi clades previously proposed, obviating laboratory-based demanding procedures. The present study constitutes an important genomic framework that could lay way for future studies on the genetic determinants underlying invasiveness and disease and population structure of H. influenzae.

Prognostic value of changes in lean and fat mass in alcoholics.

BACKGROUND & AIMS: The prognostic value of nutritional status and/or lean and fat mass assessed by dual-energy X-ray absorptiometry (DEXA) has been widely analyzed, in both alcoholics and non-alcoholics. However, the prognostic value of changes in fat and lean mass over time in alcoholics has scarcely been studied, nor has the effect of alcohol abstinence on these changes. METHODS: From an initial cohort of 113 alcoholic patients, 70 prospectively underwent two DEXA assessments six months apart. One hundred and five patients (including 66 of those who underwent two DEXA assessments) were followed up for 34.9 ± 36.4 months (median = 18 months, interquartile range = 7.25-53.75 months). During this follow-up period, 33 died (including 20 of those who had undergone a second DEXA assessment). RESULTS: Forty-two of the 70 patients undergoing a second DEXA assessment had abstained from alcohol. Of these, 69.04% (29) gained left arm lean mass, compared with only 35.71% (10 of 28) of those who had continued drinking (χ² = 7.46; p = 0.006). Similar results were observed regarding right arm lean mass (χ² = 4.68; p = 0.03) and right leg lean mass (χ² = 7.88; p = 0.005). However, no associations were found between alcohol abstinence and changes in fat parameters. Analysis by means of Kaplan-Meier curves showed that loss of total lean mass, right leg lean mass, left leg lean mass and total fat mass were all significantly associated with reduced survival. However, within 30 months of the second evaluation, significant associations were observed between changes of all parameters related to lean mass, and mortality, but no association between changes in fat parameters and mortality. CONCLUSIONS: Loss of lean mass over a period of six months after a first assessment is associated with worse prognosis in alcoholics, irrespective of whether they stop drinking during this period or not. Continued drinking is associated with greater loss of lean mass, but not with changes in fat mass.

[SAPHO syndrome. Scintigraphic and radiologic findings]. / Síndrome de SAPHO. Hallazgos gammagráficos y radiológicos.

We present the case of a 39 year old woman with a several year long history of palmoplantar pustulosis and occasional pain in the anterior wall of the thorax, lumbar spine and both sacroiliac joints. As a consequence of low grade fever, severe sternoclavicular pain and discomfort and swelling in the first left rib arches, the patient was admitted to hospital. Based on the clinical, radiologic, scintigraphic and cutaneous findings, the SAPHO syndrome was diagnosed. Bone scintigraphy is an accurate and early diagnostic test for this syndrome which also provides the advantage of being able to obtain a whole body scan.

Osteopenia assessed by body composition analysis is related to malnutrition in alcoholic patients.

Osteopenia is frequent among alcoholics. Its pathogenesis seems to be multifactorial, including ethanol intake, hormonal changes, liver cirrhosis, and malnutrition. Our objective is to determine the relative role of malnutrition on bone loss. One hundred and eighty-one male alcoholic patients, drinkers of more than 80 g ethanol/day, were included, recording data on the intensity of alcoholism, liver cirrhosis, nutritional assessment based on feeding habits, body mass index (BMI), midarm anthropometrics, subjective nutritional assessment, lean and fat mass by dual energy X-ray absorptiometry (DEXA), serum proteins and insulin growth factor Type I (IGF-I), calcitropic hormones, parathyroid hormone (PTH), osteocalcin 25OHD3, and bone mass assessed by DEXA, which was also performed in 43 healthy controls. Alcoholics showed decreased serum osteocalcin, PTH, 25OHD3, IGF-I, and bone mass. Alcoholics were frequently malnourished with decreased BMI, lean, and fat mass. The loss of bone mass was not related to the alteration of calcitropic hormones, to the intensity of alcoholism, or to the existence of liver cirrhosis, but to malnutrition. For a similar BMI, bone loss was more intense in alcoholics than in controls, especially in those with irregular feeding habits. Although cross-sectional ones, our data suggest that alcoholic osteopenia may be interpreted as a form of nutritional osteoporosis, notwithstanding the influence of other factors.

[Megaloblastic anemia in the recovery of the malnourished child]. / Anemia megaloblástica en la recuperación del ninño desnutrido

With the purpose of analyzing the evolution in weight before and after treatment of megaloblastic anemia with folic acid, we studied the clinical changes and the evolution in weight in 8 malnourished children who developed this type of anemia during their recovery. Seven out of the 8 cases were less than 20 months of age; 5 of them came from mothers who had had five or more pregnancies; in 7 a history of diarrhea of prolonged course was presented and 2 cases had been given sodium diphenylhydantoin. Increase in weight before treatment of anemia with folic acid was lesser than the one for the days following treatment in seven of the patients. Simultaneously, an important change in behavior was appreciated, without being possible to atribute this change or the increase in weight to a quantitative change favorable to the ingestion of proteins and calories.