Are GES carbapenemases underdiagnosed? An allelic discrimination assay for their accurate detection and differentiation.

GES (Guiana Extended Spectrum) carbapenemases belong to "minor class A carbapenemases" and its prevalence could be underestimated due to the lack of specific tests. The aim of this study was to develop an easy PCR method to differentiate between GES ß-lactamases with or without carbapenemase activity, based on an allelic discrimination system of SNPs that encode E104K and G170S mutations, without need of sequencing. Two pair of primers and Affinity Plus probes, labeled with different fluorophores; FAM/IBFQ and YAK/IBFQ, were designed for each one of the SNPs. This allelic discrimination assay allows to detect in real time the presence of all type of GES- ß-lactamases, being able to differentiate between carbapenemases and extended-spectrum ß-lactamase (ESBL), through a quick PCR test that avoid costly sequencing approaches and could help to decrease the current underdiagnosis of minor carbapenemases that scape of phenotypic screenings.

Antimicrobial activity of ceftolozane-tazobactam against Enterobacterales and Pseudomonas aeruginosa recovered during the Study for Monitoring Antimicrobial Resistance Trends (SMART) program in Spain (2016-2018).

OBJECTIVE: To analyse the susceptibility to ceftolozane-tazobactam and comparators in Enterobacterales and Pseudomonas aeruginosa isolates recovered from intraabdominal (IAI), urinary (UTI), respiratory (RTI) and bloodstream infection (BSI) in the SMART (Study for Monitoring Antimicrobial Resistance Trends) study. METHODS: The susceptibility of 5,351 isolates collected in 11 Spanish hospitals (2016-2018) were analysed (EUCAST-2020 criteria) by broth microdilution and were phenotypically studied for the presence of extended-spectrum beta-lactamases (ESBL). Ceftolozane-tazobactam and/or carbapenem resistant isolates were genetically characterized for ESBL and carbapenemases. RESULTS: Escherichia coli was the most frequent pathogen (49.3% IAI, 54.9% UTI, 16.7% RTI and 50% BSI), followed by Klebsiella pneumoniae (11.9%, 19.1%, 13.1% and 15.4%, respectively). P. aeruginosa was isolated in 9.3%, 5.6%, 32% and 9%, respectively. The frequency of isolates with ESBLs (2016-2017) was: 30.5% K. pneumoniae, 8.6% E. coli, 2.3% Klebsiella oxytoca and 0.7% Proteus mirabilis. Ceftolozane-tazobactam was very active against non-ESBL-(99.3% susceptible) and ESBL-(95.2%) producing E. coli being less active against K. pneumoniae (98% and 43.1%, respectively) isolates. CTX-M-15 was the most prevalent ESBL in E. coli (27.5%) and K. pneumoniae (51.9%) frequently associated with OXA-48-like carbapenemase. Overall, 93% of P. aeruginosa isolates were susceptible to ceftolozane-tazobactam, preserving this activity (>75%) in isolates resistant to other beta-lactams except in those resistant to meropenen or ceftazidime-avibactam. GES-5, PER-1, VIM-1/2 were the most prevalent enzymes in isolates resistant to ceftolozane-tazobactam. CONCLUSIONS: Ceftolozane-tazobactam showed high activity rates against isolates recovered in the SMART study although it was affected in K. pneumoniae and P. aeruginosa isolates with ESBL and/or carbapenemases.

Prospective multicentre study of rectal carriage of multidrug-resistant Enterobacteriaceae among health-care workers in Spain.

OBJECTIVES: To investigate the rectal carriage of multidrug-resistant Enterobacteriaceae (colistin-resistant, extended-spectrum ß-lactamase (ESBL) -producers and/or carbapenemase-producers) among health-care workers (HCWs) from six Spanish hospitals. METHODS: Rectal swabs from 258 HCWs, employed in intensive care units, haematology wards and clinical microbiology laboratories from six hospitals in northern Spain were studied. They were cultured in selective media for Gram-negative resistant bacteria. Detection of antimicrobial resistance genes and multilocus sequence typing were performed by PCR and further sequencing. A questionnaire including data related to risk factors of colonization/infection by resistant bacteria (age, gender, chronic diseases, immunosuppressive therapies, invasive procedures or antimicrobial treatments) was given to each participant. RESULTS: No carbapenemase-producing Enterobacteriaceae were recovered. However, 8/258 HCWs (3.1%) were positive for ESBL-producing isolates. This rate was not higher than the colonization rate previously reported in Spain for healthy people in the community. Five isolates showed high-level resistance to colistin (MICs ranging from 8 to 128 mg/L) but all of them were negative for the mcr genes tested. No statistically significant risk factors for gut colonization by ESBL-producing or colistin-resistant Enterobacteriaceae were identified among the HCWs participating in the study. CONCLUSIONS: Our data suggest that working in hospitals does not represent a risk for rectal carriage of multidrug-resistant Enterobacteriaceae.

Monitoring the antimicrobial susceptibility of Gram-negative organisms involved in intraabdominal and urinary tract infections recovered during the SMART study (Spain, 2016 and 2017).

OBJECTIVE: Continuous antimicrobial resistance surveillance is recommended by Public Health authorities. We up-dated data from the SMART (Study for Monitoring Antimicrobial Resistance Trends) surveillance study in Spain. METHODS: The antimicrobial susceptibility data and extended-spectrum beta-lactamase (ESBL) production in isolates recovered from intra-abdominal (IAI) (n=1,429) and urinary tract (UTI) (n=937) infections during the 2016- 2017 SMART study in 10 Spanish hospitals were analysed. RESULTS: Escherichia coli was the most frequently microorganism isolated (48.3% and 53.7%) followed by Klebsiella spp. (11.5% and 21.9%) in IAIs and UTIs, respectively. Figures for Pseudomonas aeruginosa were 9.0% and 6.1%, being more frequently recovered from patients with nosocomial infections. Overall, 9.9% (IAI) and 14.0% (UTI) of E. coli, Klebsiella spp. and Proteus mirabilis isolates were ESBL-producers, being Klebsiella pneumoniae (34.5%) from UTI of nosocomial origin the most frequent. ESBL-producers were higher in patients >60 years in both IAIs and UTIs. As in previous years, amikacin (96.3%-100% susceptibility), ertapenem (84.2%-100%) and imipenem (70.3%- 100%) were the most active antimicrobials tested among Enterobacterales species. The activity of amoxicillin-clavulanic, piperacillin-tazobactam, and ciprofloxacin susceptibility was lower, particularly among ESBL-producers. Ertapenem susceptibility (88.9%-100%) was retained in ESBL-E. coli isolates that were resistant to these antimicrobials but decreased (28.6%-100%) in similar isolates of K. pneumoniae. CONCLUSIONS: Continuous antimicrobial resistance surveillance from the SMART study reveals overall maintenance of ESBL-producers in Spain, although with higher presence in isolates from UTIs than from IAIs. Moreover, ertapenem activity was high in E. coli irrespective of ESBL production but decreased in K. pneumoniae, particularly among ESBL-producers.

[Spontaneous bacterial peritonitis caused by Listeria monocytogenes: eight case reports (1992-2017) and literature review]. / Peritonitis bacteriana espontánea por Listeria monocytogenes: presentación de ocho casos (1992- 2017) y revisión de la literatura.

OBJECTIVE: Spontaneous bacterial peritonitis (SBP) is a frequent and severe entity in patients with cirrhosis or ascites due to other causes. However, Listeria monocytogenes is a microorganism that has been scarcely identified as a causative agent of SBP. METHODS: In this study, a descriptive analysis of cases of L. monocytogenes SBP was carried out in our center for 26 years (1992-2017). RESULTS: A total of eight patients were diagnosed, with an average age of 58 years, with no differences in sex distribution and all of them were community acquired cases. Half of the patients had underlying liver disease, two of them active malignancies; one was undergoing continuous ambulatory peritoneal dialysis and the last one with hypertensive heart disease. Six (75%) of the patients received a third-generation cephalosporin as empirical treatment. The clinical course was favorable after receiving directed antibiotic treatment in five (62.5%) of the patients. However, three of them, under the age of 59, died. Serotyping of L. monocytogenes isolates revealed that half of them were serovar 4, two 1 / 2a and the remaining one 1 / 2c. All strains were susceptible to ampicillin, meropenem, erythromycin and cotrimoxazole. CONCLUSIONS: We conclude by emphasizing the importance of taking this etiology into account in patients with underlying liver disease and with clinical or laboratory data suggesting SBP, mainly due to the need for specific antibiotic treatment different from conventionally empirically used.

Outpatient dermatology major surgery: a 1-year experience in a Spanish tertiary hospital.

INTRODUCTION: The constant increase in the incidence of skin cancer together with the requirement for maximum exploitation of available medical resources has meant that dermatological major surgery on an outpatient basis has greatly increased in recent years. OBJECTIVE: This article reviews the practice of dermatological surgery in an outpatient setting over a 1-year period. Its purpose is to analyse the number of surgical procedures, the type of cutaneous processes treated and the kind of surgical intervention and anaesthesia used. We also assessed the percentage of cancellations, postsurgical hospital admission and postsurgical complications. Clinicopathological correlation and complete tumour removal were also evaluated. MATERIAL AND METHODS: Our clinical experience of major surgery of outpatients at the dermatology department of the Hospital Juan Canalejo in A Coruña (Spain), analysing 565 patients in a non-randomized pilot study running from January to December 2003, is presented. RESULTS: Six hundred and forty-four surgical procedures were performed on a total of 565 patients. The three main cutaneous processes treated were basal cell carcinoma (240), squamous cell carcinoma (117) and melanoma (77). The most frequent type of procedure was direct closure (346), followed by exeresis and flap (133) and partial- or full-thickness skin graft (29), nail apparatus surgery (56) and lip surgery (33). Twelve patients were admitted to hospital after surgery owing to the complexity of their operations or to complications arising during surgery. Five hundred and fifty-three patients were discharged after being kept under observation for a few hours. Seventeen patients (3%) suffered complications following surgery, which consisted of partial implant failure (six cases), infection of the surgical wound (six cases), intense pain (four cases) and haemorrhage (one case). There was a good clinicopathological correlation in 90.78% of non-melanoma skin cancers, of which 92.03% were completely removed. CONCLUSIONS: Medium and high complexity operations for dermatological processes, traditionally performed on hospitalized patients, can be conducted on an outpatient basis. This allows hospitalization costs and waiting lists to be reduced and affords the possibility of achieving better morbidity rates and medical care than in the standard hospital setting.

Acne is not a stigma of HIV infection: a review of 36 patients.

INTRODUCTION: Several diseases with follicular occlusion have been described in HIV-infected patients and can appear isolated or associated with each other in an independent disease named 'HIV-associated follicular syndrome'. Worsening of cutaneous lesions during the course of infection in patients with previous acne vulgaris, late onset or severe forms of acne have been reported in HIV-infected patients. PATIENTS AND METHODS: A prospective study of 335 HIV-infected patients was performed in our hospital. A meticulous dermatological exploration in all patients was made, and multiple data were stored into a Microsoft Access 97 program. Patients diagnosed as having acne were considered separately, and we studied their characteristics. A statistical analysis with SPSS 9.0 (Statistical Package for the Social Sciences) was performed. RESULTS: Thirty-six patients (10.8%) were diagnosed as having acne. Papulopustular lesions were the most frequent clinical presentation. Most lesions were localized on the face or on the back, and 80% of patients had mild to moderate acne; 40% of them required no treatment and the remainder was treated with topical measures, oral antibiotics and isotretinoin. 85% of cases reported the onset of acne during adolescence or before HIV infection and without any relationship with this disease. No relation between acne intensity and stage of HIV infection was observed. A multivariate analysis showed that the most important variable was age. DISCUSSION: In contrast to previous reports, no greater intensity or modifications in acne lesions were observed in our HIV-infected patients during the course of the disease. In the majority of cases, cutaneous lesions started before HIV infection, during adolescence, and the most important factor for suffering from acne was young age. In contrast to data reported in the literature, no relation of acne lesions to CD4 lymphocyte count or AIDS case criteria was found in our patients. CONCLUSIONS: Acne in HIV-infected patients has characteristics similar to those in non-HIV-infected patients. The age is the most important factor for the appearance of lesions that usually develop during adolescence. Acne lesions are not modified by HIV infection, and no relationship with the severity of HIV infection has been observed.

Metastatic tuberculous abscesses in an immunocompetent patient.

The decreased incidence of infectious diseases in developed countries may make their diagnosis difficult. Cutaneous tuberculosis is an example of this fact. A 44-year-old man presented with two painful abscesses on his lower extremities, which developed into chronic ulcers. A cutaneous biopsy revealed necrotizing granulomas in the dermis. Ziehl-Neelsen and periodic acid-Schiff stain were negative. Mantoux test was positive. Tc-99m scintigraphy showed increased uptake in the bone tissue of the left ankle and right tibiae, without direct relation to cutaneous lesions. Chest X-ray showed micronodular, apical, bilateral infiltrates, reduced volume of the right lung, and cavitation of the right superior lobe. Mycobacterium tuberculosis was grown from sputum and skin biopsy samples. Isoniazid, rifampin and pyrazinamide treatment for 2 months, followed by isoniazid and rifampin for 12 months, resulted in complete resolution. The clinical features of cutaneous tuberculosis in our patient were characteristic of tuberculous abscesses. Some uncommon findings, such as the low number of lesions, negative acid-fast resistant stains in cutaneous biopsy samples and his preserved general state of health, may be explained by a higher competence of the immune system than is usual in this clinical subset of disseminated tuberculosis. Cutaneous tuberculosis should be included in the differential diagnosis of cutaneous abscesses in immunocompetent patients.

Acne in Apert's syndrome: treatment with isotretinoin.

Apert's syndrome is an uncommon disease characterized by synostosis of extremities, vertebrae and skull. A clear association between Apert's syndrome and acne vulgaris with resistance to usual acne treatments has been described. A case of Apert's syndrome treated with oral isotretinoin with good results is reported and the pathogenic mechanisms of acne in this syndrome are discussed.

[Epidemiologic and clinical study of infection caused by hepatitis delta virus in the health service area of La Coruna]. / Estudio epidemiológico y clínico de la infección por el virus de la hepatitis delta en el área sanitaria de La Coruña.

A prospective study on 175 hepatitis HBsAg positive patients was carried out between March 1986 and June 1987 in order to study the epidemiological and clinical aspects of the HVD infection. The global prevalence of this infection was 19.42%, with a statistically relevant predominance amongst intravenous drug addicts (IVDA). Moreover, in most cases it was associated with liver disease (91.17%). Chronic liver disease had a greater incidence amongst HVD positive infected patients and presented more severe clinical and biochemical aspects than other chronic liver diseases without HVD infection. The HVD infection was confirmed in 39.4% of the HBs Ag positive acute hepatitis, being in the majority of the cases delta coinfections with a clinical and biochemical evolution similar to the hepatitis caused by the B virus only. The overinfections evolved to cronicity in all cases. No delta infection occurred amongst immunodepressed patients, and there was an incidence amongst hemophiliacs of 40%. In view of our results, we may conclude that in our community the parenteral route is the main mechanism for the transmission of HVD.

[Comparative study on the clinico-serologic course of acute hepatitis B and hepatitis B with delta co-infection]. / Estudio comparativo sobre la evolución clínico-serológica de las hepatitis agudas B y las hepatitis B con coinfección delta.

The clinical and serological course of 35 instances of hepatitis B (22 hepatitis B alone and 13 with delta co-infection) was comparatively and prospectively studied for a mean follow-up period of 7.19 +/- 2.69 months. There were no significant differences between the clinical presentation and course between both types of patients. There were not, either, outstanding differences in the mean bilirubin and transaminase levels between both types of hepatitis, although the patients with delta co-infection initially showed a more pronounced cholestatic pattern than did patients with hepatitis B alone (p less than 0.05). In hepatitis B with delta co-infection, the serological markers of activity and/or replication of hepatitis B virus (HBV) showed an earlier serum clearance than in patients with hepatitis B alone, basically in relation with the "e" antigen of HBV. Serum measurement of hepatitis delta virus (HDV) antigen by means of ELISA was shown to be a high sensitivity test (69.23%) for the diagnosis of acute HDV hepatitis, which was carried out in an early stage since the development of symptoms.