Reliability of a Screening Method Using Antibiotic Disks to Detect Carbapenemases in Glucose-Nonfermenting Gram-Negative Microorganisms From Clinical Samples of a Regional Hospital in Southeastern Spain.

BACKGROUND: Infections by glucose-nonfermenting gram-negative bacilli (NFGNB) pose a major public health problem due to multiresistance to beta-lactam antibiotics, especially plasmid-borne carbapenemases. Their detection by microbiology laboratories is challenging, and there is a need for easy-to-use and reliable diagnostic techniques. Our objective was to evaluate an in-house screening method to presumptively detect carbapenemases in NFGNB in a simple and clinically useful manner. METHODS: The study included 175 NFGNB isolates from urinary, respiratory, and rectal samples. In a triple assay, isolates were incubated at 37°C for 24 h on three solid-culture media: MacConkey II Agar, 5% Sheep Blood Columbia Agar and Mueller Hinton II Agar; meropenem (MEM) and cefepime (FEP) disks were employed for screening. Studies were then performed on the inhibition halo diameter, scanning effects, and the appearance of mutant colonies, which were compared with those observed using the colorimetric Neo-Rapid CARB Kit and immunochromatography (NG5-Test Carba and K-Set for OXA-23). Receiver operating characteristic curves were constructed for these data. RESULTS: Carbapenemases were expressed by 79/175 (45.1%): 19 Pseudomonas aeruginosa and 60 Acinetobacter baumannii. Optimal inhibition halo diameter cutoffs to detect this resistance on 5% sheep blood agar were as follows: 6 mm (MEM) and 6.5 mm (FEP) for P. aeruginosa (in the absence of scanning effects and mutations) and 10.5 mm (MEM) and 16 mm (FEP) for A. baumannii (even in the presence of scanning effects). CONCLUSION: The combined utilization of MEM and FEP antibiotic disks in 5% sheep blood agar, measuring their inhibition haloes, offers an effective method to predict the presence of carbapenemases as resistance mechanism in P. aeruginosa and A. baumannii.

Concerning the presumptive identification of Candida kefyr on Uriselect™4 agar / Sobre la identificación presuntiva de Candida kefyr en agar Uriselect™4

Introduction. Non-albicans Candida species, such as Candida kefyr, are emerging pathogens. Chromogenic media are highly useful for the diagnosis of urinary tract infections (UTIs). The aim was to describe the behavior of this specie on a non-specific chromogenic medium. Material and methods. A retrospective study of cases of candiduria detected in the Microbiology laboratory of the Virgen de las Nieves Hospital in Granada (Spain) between 2016 and 2021 (N=2,130). Urine samples were quantitatively seeded on non-selective UriSelect™4 chromogenic agar. Results. Between 2016 and 2021, C. kefyr was the seventh most frequent Candida species responsible for candiduria in our setting (n=15). The macroscopic appearance of C. kefyr colonies, punctiform and bluish, allowed the direct identification of these microorganisms. Conclusions. This study provides the first description of the specific behavior of C. kefyr on UriSelect™4 agar, which differentiates it from other Candida species based on its enzymatic characteristics. (AU)

Introducción. Las especies de Candida no-albicans, como Candida kefyr, son patógenos emergentes. Los medios cromogénicos son muy útiles para el diagnóstico de infecciones del tracto urinario (ITU). El objetivo era describir el comportamiento de esta especie en un medio cromogénico no específico. Material y métodos. Estudio retrospectivo de casos de candiduria detectados en el laboratorio de Microbiología del Hospital Virgen de las Nieves de Granada (España) entre 2016 y 2021 (N=2.130). Las muestras de orina se sembraron cuantitativamente en agar cromogénico no selectivo Uri Select™4. Resultados. C. kefyr fue la séptima especie de Candida responsables de la candiduria en nuestro medio (n = 15). El aspecto macroscópico de las colonias de C. kefyr, puntiformes y azuladas, permitió su identificación presuntiva directamente. Conclusiones. Este estudio proporciona la primera descripción del comportamiento específico de C. kefyr en agar Uri Select™4, que lo diferencia de otras especies de Candida en función de sus características enzimáticas. (AU)

Preliminary reading of antibiogram by microdilution for clinical isolates in urine culture.

PURPOSE: We evaluated a modification of automated antibiograms in urine cultures designed to facilitate the early interpretation of minimum inhibitory concentrations (MICs) and accelerate the targeted treatment of urinary tract infections (UTIs), METHODS: A prospective study was conducted of 309 isolates (219 Enterobacteriaceae, 75 Enterococcus spp., and 15 non-fermenting Gram-negative bacilli (NFGNB), and a retrospective study of 9 carbapenemase-producing clinical isolates from urine cultures. Colonies grown on conventional isolation plates were inoculated in MicroScan Walkaway system panels and incubated for 7 h, using a MicroScan AutoScan-4 plate reader for preliminary MIC determination by turbidimetry. Resulting antibiograms were compared with definitive antibiograms obtained after incubation for 17 h. RESULTS: Preliminary and definitive readings were concordant for 86.7% of Gram-positive cocci isolates (65/75), 61.6% of Enterobacteriaceae (135/219), and 53.3% of NFGNB. The agreement rate was greater than 90% for most antimicrobials against Gram-positive cocci (94.7% or more) and Enterobacteriaceae, (97.2% or more for 10 of 17 antibiotics) except with nitrofurantoin (89%). The agreement rate was 86.7% or more for most antibiotics against NFGNB apart from piperacillin/tazobactam, aztreonam, amikacin, and ciprofloxacin. Gram-negative bacilli showed the highest differences in MIC values between preliminary and definitive readings. CONCLUSIONS: A preliminary antibiogram reading may be useful in urine cultures to reduce the delay before targeted antibiotherapy, especially against Enterobacteriaceae and Gram-positive cocci, but not in cases of carbapenemase-producing NFGNB. Further local studies are warranted to evaluate the usefulness of this approach in relation to resistance rates.

Antibiotic resistances of Pseudomonas aeruginosa and Acinetobacter baumannii in urine cultures: experience in a hospital of Southeast Spain.

The objectives of this study were to perform a systematic review of publications between 2010 and 2021 on the antibiotic resistance of Pseudomonas aeruginosa and Acinetobacter baumannii from urinary tract infections and to analyze changes over time in hospital urine cultures from 2016 through 2021. The literature was searched, and a retrospective cross-sectional descriptive study was performed in the hospital. Out of 21 838 positive urine cultures, 3.86% were due to P. aeruginosa and 0.44% were due to A. baumannii. For P. aeruginosa, lower resistance rates were observed to virtually all tested antibiotics than were obtained in the systematic review, and the present series of hospital samples showed an in vitro resistance rate <10% to ceftazidime, cefepime, meropenem, piperacillin-tazobactam, amikacin, tobramycin, and colistin. For A. baumannii, the resistance rates to almost all antibiotics were higher in the present series than in the systematic review, being lowest to colistin (10%). Both microorganisms show reduced in vitro susceptibility to some antibiotics during the years of the COVID-19 pandemic in comparison to previous years. In our setting, both piperacillin-tazobactam and meropenem can be recommended for the empirical treatment of UTIs by P. aeruginosa, whereas only colistin can be recommended for UTIs by A. baumannii.

Shigelosis atendidas en el servicio de urgencias de un hospital regional del sudeste de España: Desde su presencia a la multirresistencia / Shigellosis attended in the emergency department of a regional hospital in southeastern Spain: from their presence to multiresistance

Introduction. In the bacterial etiology of severe acute infectious diarrhea, except that caused by Clostridioides difficile, most of them have an invasive character and antibiotic treatment will be necessary in specific situations. Shigella is a classic pathogen, in which it is crucial to know the sensitivity to different classic and alternative antimicrobials. The objective of this work was to analyze the presence of shigellosis and the rate of antibiotic resistance.Methods. A descriptive-retrospective study of the reports of shigellosis of stool cultures issued between January 2016 and April 2022 was conducted.Results. A total of 34 episodes (16 -47.1%- by Shigella sonnei) were observed, as of 2018. There were only 2 pediatric cases. The overall resistance rate to azithromycin, trimethoprim-sulfamethoxazole and ciprofloxacin was 52.9%, 64.7% and 44.1%, respectively. 26.5% were resistant to the 3 groups of antibiotics. There was a higher rate of resistance for S. sonnei. The emergence of resistance to cephalosporins in recent years stands out. Episodes of multidrug-resistant shigellosis were detected between 2020 (1 by S. flexneri) and 2022 (4 by S. sonnei).Conclusions. The episodes of shigellosis are emerging in our environment with a higher rate of multi-resistance. In this context, current empirical treatments for acute enteroinvasive enteritis are at risk of failure, if necessary.

Introducción: En la etiología bacteriana de la diarrea infecciosa aguda grave, exceptuando la causada por Clostridioides difficile, la mayor parte presentan un carácter invasor y el tratamiento antibiótico será preciso en situaciones concretas. Shigella es un patógeno clásico, en el que es crucial conocer la sensibilidad a distintos antimicrobianos clásicos y alternativos. El objetivo de este trabajo fue analizar la presencia de shigelosis y la tasa de resistencia a los antibióticos. Métodos: Se realizó un estudio descriptivo-retrospectivo de los informes de shigelosis de los coprocultivos emitidos entre enero de 2016 y abril de 2022. Resultados: Se observó un total de 34 episodios (16-47,1%- por Shigella sonnei), a partir del 2018. Sólo hubo 2 casos pediátricos. La tasa de resistencia global a azitromicina, trimetoprim-sulfametoxazol y ciprofloxacino fue de 52,9%, 64,7% y 44,1%, respectivamente. El 26,5% fueron resistentes a los 3 grupos de antibióticos. Hubo mayor tasa de resistencia por S. sonnei. Destaca la aparición de resistencia a cefalosporinas en los últimos años. Los episodios de shigelosis multirresistente se detectaron entre 2020 (1 por S. flexneri) y 2022 (4 por S. sonnei). Conclusiones: Los episodios de shigelosis importada están emergiendo en nuestro medio con una mayor tasa de multirresistencia. En este contexto, los tratamientos empíricos actuales para las enteritis agudas enteroinvasivas corren el riesgo de fracasar, en caso de ser necesarios. (AU)

Antibiotic Resistances of Enterobacteriaceae with Chromosomal Ampc in Urine Cultures: Review and Experience of a Spanish Hospital.

The Enterobacteriaceae Citrobacter freundii, Enterobacter cloacae, Klebsiella aerogenes, Morganella morganii, Providencia stuartii, and Serratia marcescens (CESPM group) produce numerous urinary tract infections (UTIs) which are difficult to treat due to their high multiresistance rate. The objectives of this study were to carry out a systematic review of antibiotic resistances by UTIs and to determine changes over time in urine cultures from a reference hospital in southern Spain. The literature was searched for European data on the resistance rates of each microorganism, and a retrospective cross-sectional descriptive study was performed in samples with suspicion of UTI from patients in Virgen de las Nieves University Hospital (Granada, Spain) between 2016 and the first half of 2021. Among 21,838 positive urine cultures, 1.85% were caused by E. cloacae, 0.77% by M. Morganii, 0.65% by K. aerogenes, 0.46% by C. freundii, 0.29% by P stuartii, and 0.25% by S. marcescens. The lowest resistance rates by microorganism were: E. cloacae to amikacin (3.47%) and imipenem (5.28%); M. morganii to piperacillin-tazobactam (1.79%), cefepime (4.76%), and tobramycin (7.74%); K. aerogenes to tobramycin (3.55%), gentamicin (4.25%), trimethoprim-sulfamethoxazole (4.96%), imipenem (5.75%), and cefepime (6.43%); C. freundii to imipenem (no resistance), nitrofurantoin (1.96%), fosfomycin (2.80%), and ertapenem (6.12%); P. stuartii to cefepime (3.28%) and ceftazidime (3.28%); and S. marcescens to gentamicin (1.8%), ciprofloxacin (3.64%), cefepime (3.70%), piperacillin-tazobactam (3.70%), and trimethoprim-sulfamethoxazole (5.45%). In our setting, CESMP Enterobacteriaceae showed the lowest resistance to piperacillin-tazobactam, cefepime, imipenem, gentamicin, and colistin, which can therefore be recommended for the empirical treatment of UTIs. The COVID-19 pandemic may have had a clinical impact in relation to the increased resistance of E. cloacae and M. morgani to some antibiotics.

Systematic Review of Plasmid AmpC Type Resistances in Escherichia coli and Klebsiella pneumoniae and Preliminary Proposal of a Simplified Screening Method for ampC.

Beta-lactamase (BL) production is a major public health problem. Although not the most frequent AmpC type, AmpC-BL is increasingly isolated, especially plasmid AmpC-BL (pAmpC-BL). The objective of this study was to review information published to date on pAmpC-BL in Escherichia coli and Klebsiella pneumoniae, and on the epidemiology and detection methods used by clinical microbiology laboratories, by performing a systematic review using the MEDLINE PubMed database. The predictive capacity of a screening method to detect AmpC-BL using disks with cloxacillin (CLX) was also evaluated by studying 102 Enterobacteriaceae clinical isolates grown in CHROMID ESBL medium with the addition of cefepime (FEP), cefoxitin (FOX), ertapenem (ETP), CLX, and oxacillin with CLX. The review, which included 149 publications, suggests that certain risk factors (prolonged hospitalization and previous use of cephalosporins) are associated with infections by pAmpC-BL-producing microorganisms. The worldwide prevalence has increased over the past 10 years, with a positivity rate ranging between 0.1 and 40%, although AmpC was only detected when sought in a targeted manner. CMY-2 type has been the most prevalent pAmpC-BL-producing microorganism. The most frequently used phenotypic method has been the double-disk synergy test (using CLX disks or phenyl-boronic acid and cefotaxime [CTX] and ceftazidime) and the disk method combined with these inhibitors. In regard to screening methods, a 1-µg oxacillin disk with CLX showed 88.9% sensitivity, 100% specificity, 100% positive predictive value (PPV), 98.9% negative predictive value (NPV), and 98.9% validity index (VI). This predictive capacity is reduced with the addition of extended-spectrum beta-lactamases, showing 62.5% sensitivity, 100% specificity, 100% PPV, 93.5% NPV, and 94.1% VI. In conclusion, there has been a worldwide increase in the number of isolates with pAmpC-BL, especially in Asia, with CMY-2 being the most frequently detected pAmpC-BL-producing type of microorganism. Reduction in its spread requires routine screening with a combination of phenotypic methods (with AmpC inhibitors) and genotypic methods (multiplex PCR). In conclusion, the proposed screening technique is an easy-to-apply and inexpensive test for the detection of AmpC-producing isolates in the routine screening of multidrug-resistant microorganisms.

Misidentification of Phocaeicola (Bacteroides) dorei in two patients with bacteremia.

Phocaeicola (Bacteroides) dorei is a Gram-negative anaerobic bacillus that is rarely isolated from human specimens. Its accurate identification can be hampered by its close taxonomic relationship with Bacteroides vulgatus. We report on two patients with bacteremia due to P. (B.) dorei, which was initially identified as B. vulgatus by MALDI-TOF MS.

Predictive Factors for Pulmonary Homograft Dysfunction After Ross Surgery: A 20-Year Follow-up.

BACKGROUND: We studied the determinants of hemodynamics and analyzed the incidence, risk factors, and clinical impact of pulmonary homograft dysfunction following Ross surgery, after a 20-year follow-up at our referral center. METHODS: From 1997 to 2017, a total of 142 patients underwent surgery using the Ross procedure. The development of moderate-severe stenosis (peak transhomograft pressure gradient 36 mm Hg or greater) and surgical or percutaneous Ross homograft reinterventions were evaluated by echocardiography in the immediate postoperative period and at annual intervals. RESULTS: After 20 years of follow-up, 31% of patients had moderate-severe homograft stenosis, and 9.1% had had to undergo one or two reinterventions, of which, six were valve replacements and seven were percutaneous interventions. At 1, 5, and 20 years, 89.4%, 74.6%, and 69% of these patients, respectively, were free from moderate-severe stenosis; and 99.3%, 95.7%, and 90.9%, respectively, had freedom from homograft reintervention. The pediatric group had a higher risk factor for homograft stenosis (hazard ratio 3.70; 95% confidence interval, 1.56 to 7.20, P = .002), whereas donor age behaved as a protective factor (hazard ratio 0.98; 95% confidence interval, 0.95 to 0.99; P = .044). Pulmonary homograft stenosis tended to appear in the first year (10.6%) or at 5 years (25.4%). CONCLUSIONS: Pulmonary homografts implanted in the Ross procedure offer satisfactory long-term results, but the level of homograft dysfunction is not negligible. Young recipient and donor age were associated with a higher rate of homograft stenosis during follow-up. Moreover, homograft dysfunction usually occurred during the first few years of follow-up, and may have been related to immune responses.

New Simplified Diagnostic Decision Trees for the Detention of Metabolic Syndrome in the Elderly.

BACKGROUND: A new simplified method for the detention of metabolic syndrome (MetS) is proposed using two variables (anthropometric and minimally invasive). METHODS: A study of MetS prevalence was made on a sample of 361 older people. The anthropometric variables analyzed were: blood pressure, body mass index, waist circumference (WC), waist-height ratio, body fat percentage, and waist-hip ratio. A crude and adjusted binary logistic regression was performed, and receiver operating characteristic curves were obtained for determining the predictive capacity of those variables. For the new detection method, decision trees were employed using automatic detection by interaction through Chi-square. RESULTS: The prevalence of the MetS was of 43.7%. The final decision trees uses WC and basal glucose (BG), whose cutoff values were: for men, WC ≥ 102.5 cm and BG > 98 mg/dL (sensitivity = 67.1%, specificity = 90.3%, positive predictive value = 85%, validity index = 79.9%); and for women, WC ≥ 92.5 cm and BG ≥ 97 mg/dL (sensitivity = 65.9%, specificity = 92.7%, positive predictive value = 87.1%, validity index = 81.3%). In older women the best predictive value of MetS was a WC of 92.5 cm. CONCLUSIONS: It is possible to make a simplified diagnosis of MetS in older people using the WC and basal capillary glucose, with a high diagnostic accuracy and whose use could be recommended in the resource-poor health areas. A new cutting point in older women for the WC should be valued.

Waist to Height Ratio and Metabolic Syndrome as lung dysfunction predictors.

Metabolic Syndrome (MetS) has been related to pulmonary diseases but its relationship with lung age has not been sufficiently studied. In addition, anthropometric variables have been associated with pulmonary dysfunction, highlighting the waist-to-height ratio (WHtR). The aim was to evaluate the relationship between MetS and: lung age, anthropometric variables and the alteration of lung function. A cross-sectional study was carried out in 1901 workers, evaluating lung function through lung age (Morris & Temple equation) and spirometric values. The diagnosis of MetS was based on the harmonized criteria. We measured anthropometric variables (WHtR, waist circumference, body mass index, waist to hip ratio), blood pressure and biochemical variables (glucose, cholesterol total, HDL, triglycerides). Workers suffering from MetS showed an accelerated lung aging (59.4 ± 18.7 years vs 49 ± 18.4 years). The WHtR ≥ 0.55 was significantly related to an increase in lung age (ß = 6.393, p < 0.001). In addition, a significant linear trend was found between clinical categories of WHtR and lung dysfunction, restrictive and mixed pattern. MetS caused an accelerated lung aging and favored the presence of restrictive lung impairment. In addition, WHtR ≥ 0.55 has been shown as the best predictor for pulmonary health.