Carbapenemase-Producing Klebsiella pneumoniae in COVID-19 Intensive Care Patients: Identification of IncL-VIM-1 Plasmid in Previously Non-Predominant Sequence Types.

During the COVID-19 pandemic, intensive care units (ICUs) operated at or above capacity, and the number of ICU patients coinfected by nosocomial microorganisms increased. Here, we characterize the population structure and resistance mechanisms of carbapenemase-producing Klebsiella pneumoniae (CP-Kpn) from COVID-19 ICU patients and compare them to pre-pandemic populations of CP-Kpn. We analyzed 84 CP-Kpn isolates obtained during the pandemic and 74 CP-Kpn isolates obtained during the pre-pandemic period (2019) by whole genome sequencing, core genome multilocus sequence typing, plasmid reconstruction, and antibiotic susceptibility tests. More CP-Kpn COVID-19 isolates produced OXA-48 (60/84, 71.4%) and VIM-1 (18/84, 21.4%) than KPC (8/84, 9.5%). Fewer pre-pandemic CP-Kpn isolates produced VIM-1 (7/74, 9.5%). Cefiderocol (97.3-100%) and plazomicin (97.5-100%) had the highest antibiotic activity against pandemic and pre-pandemic isolates. Sequence type 307 (ST307) was the most widely distributed ST in both groups. VIM-1-producing isolates belonging to ST307, ST17, ST321 and ST485, (STs infrequently associated to VIM-1) were detected during the COVID-19 period. Class 1 integron Int1-blaVIM-1-aac(6')-1b-dfrB1-aadAI-catB2-qacEΔ1/sul1, found on an IncL plasmid of approximately 70,000 bp, carried blaVIM-1 in ST307, ST17, ST485, and ST321 isolates. Thus, CP-Kpn populations from pandemic and pre-pandemic periods have similarities. However, VIM-1 isolates associated with atypical STs increased during the pandemic, which warrants additional monitoring and surveillance.

Increased risk of group B streptococcal sepsis and meningitis in HIV-exposed uninfected infants in a high-income country.

The purpose of this study is to compare group B Streptococcus (GBS) infection incidence in HIV-exposed uninfected (HEU) and HIV-unexposed (HU) infants in a Spanish cohort. We conducted a retrospective study in 5 hospitals in Madrid (Spain). Infants ≤ 90 days of life with a GBS infection were included from January 2008 to December 2017. Incidence of GBS infection in HEU and HU children was compared. HEU infants presented a sevenfold greater risk of GBS infection and a 29-fold greater risk of GBS meningitis compared to HU, with statistical significance. Early-onset infection was tenfold more frequent in HEU children, with statistical significance, and late-onset infection was almost fivefold more frequent in the HUE infants' group, without statistical significance. CONCLUSION: HEU infants presented an increased risk of GBS sepsis and meningitis. One in each 500 HEU infants of our cohort had a central nervous system infection and 1 in each 200, a GBS infection. Although etiological causes are not well understood, this should be taken into account by physicians when attending this population. WHAT IS KNOWN: • HIV-exposed uninfected infants are at higher risk of severe infections. • An increased susceptibility of these infants to group B Streptococcus infections has been described in low- and high-income countries, including a higher risk of meningitis in a South African cohort. WHAT IS NEW: • Group B Streptococcal meningitis is more frequent in HIV-exposed uninfected infants also in high-income countries. • Physicians should be aware of this increased risk when attending these infants.

Mobile colistin resistance (MCR), extended-spectrum beta-lactamase (ESBL) and multidrug resistance monitoring in Escherichia coli (commensal and pathogenic) in pig farming: need of harmonized guidelines and clinical breakpoints.

Current data on antimicrobial resistance in pig production is essential for the follow-up strategic programs to eventually preserve the effectiveness of last-resort antibiotics for humans. Here, we characterized 106 Escherichia coli recovered in routine diagnosis (2020-2022) from fecal sample pigs, belonging to 74 Spanish industrial farms, affected by diarrhea. The analysis of virulence-gene targets associated with pathotypes of E. coli, determined 64 as pathogenic and 42 as commensal. Antimicrobial susceptibility testing (AST) performed by minimal inhibitory concentration (MIC) assay, was interpreted by applying breakpoints/cut-off values from the different standards EUCAST/TECOFF 2022, CLSI VET ED5:2020, and CASFM VET2020. Comparisons taking EUCAST as reference exhibited moderate to high correlation except for enrofloxacin, neomycin, and florfenicol. Of note, is the lack of clinical breakpoints for antibiotics of common use in veterinary medicine such as cefquinome, marbofloxacin, or florfenicol. AST results determined multidrug resistance (MDR) to ≥3 antimicrobial categories for 78.3% of the collection, without significant differences in commensal vs pathogenic isolates. Plasmid-mediated mobile colistin resistance gene (mcr) was present in 11.3% of 106 isolates, all of them pathogenic. This means a significant decrease compared to our previous data. Furthermore, 21.7% of the 106 E. coli were ESBL-producers, without differences between commensal and pathogenic isolates, and mcr/ESBL genes co-occurred in 3 isolates. Phylogenetic characterization showed a similar population structure (A, B1, C, D, and E), in both commensal and pathogenic E. coli, but with significant differences for B1, C, and E (38.1 vs 20.3%; 19 vs 1.6%; and 7.1 vs 25%, respectively). Additionally, we identified one B2 isolate of clone O4:H5-B2-ST12 (CH13-223), positive for the uropathogenic (UPEC) status, and in silico predicted as human pathogen. We suggest that a diagnosis workflow based on AST, detection of mcr and ESBL genes, and phylogenetic characterization, would be a useful monitoring tool under a "One-Health" perspective.

Determining Time to Positivity of Blood Cultures in a Neonatal Unit.

Blood cultures are the gold standard for detecting bacteremia. We have studied the time to positivity of blood cultures in our neonatal unit to reduce antibiotic doses in patients with a negative blood culture. Empirical antibiotic treatment of neonatal sepsis could be withdrawn 24 hours after obtaining blood cultures.

Clustering Analysis of the Multi-Microbial Consortium by Lactobacillus Species Against Vaginal Dysbiosis Among Ecuadorian Women.

The vaginal microbiota plays vital protection in women. This probiotic activity is caused not only by individual Lactobacillus species but also by its multi-microbial interaction. However, the probiotic activity promoted by multi-microbial consortia is still unknown. The aim of this study was the individual and collective analysis on the prevalence of five vaginal lactobacilli (Lactobacillus iners, Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, and Lactobacillus acidophilus) among healthy women and women with bacterial vaginosis (BV) or aerobic vaginitis (AV). PCR assays were realized on 436 vaginal samples from a previous study. Chi-square, univariable, and multivariable logistic regression analyses with the Benjamini-Hochberg adjustment evaluated associations between these lactobacilli and vaginal microbiota. Multi-microbial clustering model was also realized through Ward's Minimum Variance Clustering Method with Euclidean squared distance for hierarchical clustering to determine the probiotic relationship between lactobacilli and vaginal dysbiosis. Concerning the individual effect, L. acidophilus, L. jensenii, and L. crispatus showed the highest normalized importance values against vaginal dysbiosis (100%, 79.3%, and 74.8%, respectively). However, only L. acidophilus and L. jensenii exhibited statistical values (p = 0.035 and p = 0.050, respectively). L. acidophilus showed a significant prevalence on healthy microbiota against both dysbioses (BV, p = 0.041; and AV, p = 0.045). L. jensenii only demonstrated significant protection against AV (p = 0.012). Finally, our results evidenced a strong multi-microbial consortium by L. iners, L. jensenii, L. gasseri, and L. acidophilus against AV (p = 0.020) and BV (p = 0.009), lacking protection in the absence of L. gasseri and L. acidophilus.

The effectiveness of an online training program in a nursing unit: extraction of blood cultures. / Efectividad de un programa de formación online en una unidad de enfermería: extracción de hemocultivos.

OBJECTIVE: To assess the effectiveness of an online training platform for procedures among nurses in an internal medicine unit to reduce the number of contaminated blood cultures. METHOD: This was a quasi-experimental pre-post intervention parallel group study. The sample consisted of internal medicine nurses in a tertiary hospital who participated in an online training program about blood culture extraction technique. Knowledge about the technique was measured pre- and post-intervention. Additionally, the study compared the number of blood cultures taken 6 months before and 3 months after the intervention. RESULTS: Forty-eight nurses participated. Pre-intervention knowledge was homogeneous among both groups, improving significantly after the online training program (p=0.0001). The blood cultures taken prior to the training showed contamination levels above international standards; post-intervention, contamination levels fell by up to 3% in the intervention group. CONCLUSION: The educational intervention using the digital platform increased knowledge about the procedure and its application in clinical practice.

Micropropagation of Solanum quitoense var. quitoense by apical bud, petiole and hypocotyl culture.

The development of in vitro propagation methods can improve the current commercial use and conservation of plants like naranjilla (Solanum quitoense), a distinctive Andean crop and key emerging agricultural product. In the present study, we report in vitro culture protocols for naranjilla apical buds, hypocotyls and petioles. In apical bud culture, MS medium supplemented with 0.10 mg l-1 1-naphtaleneacetic acid (NAA) produced longer plantlets with greater number of leaves. Hypocotyl culture yielded higher number of shoots when using older explants in MS medium supplemented with different combinations of NAA, 6-benzylaminopurine (BAP) and gibberellic acid (GA3). Petiole culture produced a significantly higher number of shoots per explant, with more abundant and bigger leaves, when using MS medium supplemented with 0.02 mg l-1 NAA, 4.50 mg l-1 BAP and 1.00 mg l-1 GA3. A factorial analysis reveals that the interaction between GA3 and NAA/BAP plays an important role in shoot regeneration. These results provide new tools for the in vitro regeneration of naranjilla plants, improving on previously reported protocols for this species by using alternative explant types and regeneration protocols.

Risk factors associated with Clostridium difficile infection in adult oncology patients.

PURPOSE: Clostridium difficile infection (CDI) prevention is particularly important for cancer patients, because diarrhea often results in dose reductions or delays of chemotherapy or radiotherapy. We conducted this study to better ascertain the incidence, susceptibility, and risk factors for CDI in cancer patients receiving chemotherapy at our hospital. METHODS: We performed a retrospective study among adult cancer patients admitted at "12 de Octubre" University Hospital between January 2009 through April 2013 who were diagnosed with diarrhea. Inpatient data were available on hospital medical records. We screened by immunochromatography system detecting glutamate dehydrogenase antigen, and C. difficile toxins A and B. Later, a polymerase chain reaction for detecting toxin B gene was performed. RESULTS: A total of 225 patients were included in the study, and 39 of them (17.3 %) were diagnosed with CDI. Type of tumor significantly differed between CDI patients, thus relative risk in each type of cancer was calculated after adjusting for age, antibiotic exposure, corticosteroid, and proton-pump inhibitor use. Patients with gastrointestinal tumors were less prone to CDI. Conversely, breast cancer patients have a greater predisposition to CDI. Antibiotic treatment was found to be associated with an increasing risk for CDI in breast cancer patients. Curiously, exposure to proton-pump inhibitors appeared protective in our cohort, except for lung cancer patients. However, we have not been able to find an association between a particular type of chemotherapy and CDI. CONCLUSIONS: We underscore the urgent need for early recognition and diagnosis of CDI in cancer patients. Our findings indicate a probable association between antibiotic use and CDI incidence, at least in certain cancer, such as breast cancer.

Algorithm proposal based on the C. Diff Quik Chek Complete ICT device for detecting Clostridium difficile infection

Objective: To assess a new immunochromatography (ICT) test that detects glutamate dehydrogenase (GDH) antigen and Clostridium difficile toxin A/B simultaneously, and to propose an algorithm for the diagnosis of C. difficile infection (CDI) based on this test. Methods We analysed 970 stool samples. Discrepant results between GDH and toxin A/B were resolved using toxigenic culture as the reference. Results This test enabled us to obtain a conclusive result in <30min in 93.8% of the samples. Among the discrepant results (GDH (+)/Toxin A/B (-)), 41.7% (25/60) were found to be toxigenic C. difficile by toxigenic culture. Conclusion This test has a high sensitivity and specificity for the diagnosis of CDI (AU)

Objetivo: Evaluar una nueva prueba inmunocromatográfica que detecta el antígeno glutamato deshidrogenasa (GDH) y la toxina A/B de Clostridium difficile simultáneamente y proponer un algoritmo para el diagnóstico de infección por C. difficile (ICD) basado en esta prueba. Métodos: Se analizaron 970 muestras. Las discrepancias entre GDH y toxina A/B se resolvieron utilizando el cultivo toxigénico como método de referencia. Resultados: Esta prueba permitió obtener el resultado del 93,8% de las muestras en < 30 minutos. El 41,7%(25/60) de las muestras discrepantes (GDH (+)/Toxina A/B (-)) fueron C. difficile toxigénicos, mediante cultivo toxigénico. Conclusión: Esta prueba es sensible y específica para el diagnóstico de ICD (AU)

Algorithm proposal based on the C. Diff Quik Chek Complete ICT device for detecting Clostridium difficile infection.

OBJECTIVE: To assess a new immunochromatography (ICT) test that detects glutamate dehydrogenase (GDH) antigen and Clostridium difficile toxin A/B simultaneously, and to propose an algorithm for the diagnosis of C. difficile infection (CDI) based on this test. METHODS: We analysed 970 stool samples. Discrepant results between GDH and toxin A/B were resolved using toxigenic culture as the reference. RESULTS: This test enabled us to obtain a conclusive result in <30min in 93.8% of the samples. Among the discrepant results (GDH (+)/Toxin A/B (-)), 41.7% (25/60) were found to be toxigenic C. difficile by toxigenic culture. CONCLUSION: This test has a high sensitivity and specificity for the diagnosis of CDI.

¿Qué tratamiento empírico es el más adecuado en pacientes con uretritis? / Which is the best empirical treatment in patients with urethritis?

Objetivo. Conocer el tratamiento empírico más adecuado de uretritis, en pacientes de la zona Centro de Madrid. Métodos. Se analizó el exudado uretral de 2.021 hombres entre Enero 2003-Diciembre 2007. Además de los cultivos tradicionales se determinó la presencia de Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, Trichomonas vaginalis y Herpes simplex. La sensibilidad de Neisseria gonorrhoeae y Haemophilus spp. se realizó mediante el método de difusión en disco y la sensibilidad de U. urealyticum mediante Micoplasma IST2. Resultados. El porcentaje de muestras positivas fue de 30,6%. Los microorganismos aislados más frecuentemente fueron: U. urealyticum 9,9%, N. gonorrhoeae 7,4%, C. trachomatis 5,1% y Haemophilus spp 3,8%. La resistencia de N. gonorrhoeae en el primer periodo fue: penicilina 11,8%, tetraciclina 5,9%, ciprofloxacino 8,8% y presencia de betalactamasas 11,8%. En el segundo periodo: penicilina 9,7%, amoxicilina/ácido clavulánico 1,4%, tetraciclina 8,3%, ciprofloxacino 23,6% y presencia de betalactamasas 10,5%. La resistencia a ciprofloxacino en no HSH (hombres que tienen relaciones sexuales con hombres) 20% y en HSH 56,2% (p<0,05). La resistencia de Haemophilus spp en el primer periodo fue: ampicilina 38,2%, amoxicilina/ácido clavulánico 8,8%, claritromicina 35,3%, cotrimoxazol 64,7%, cefuroxima 5,9%, ciprofloxacino 8,8%, tetraciclina 12,1% y presencia de betalactamasas 26,5%. En el segundo periodo: presencia de betalactamasas 41,9%, ampicilina 53,1%, amoxicilina/ácido clavulánico 9,4%, cefuroxima 9,4%, claritromicina 18,7%, tetraciclina 34,4%, ciprofloxacino 15,6% y cotrimoxazol 68,7%. La resistencia de U. urealyticum fue: 80,7% ciprofloxacino, 32,4% ofloxacino, 17,5% eritromicina, 9,6% azitromicina, 3,5% tetraciclina y 0,8% doxiciclina. Conclusiones. N. gonorrhoeae presentó mayor resistencia a tetraciclina y ciprofloxacino en el segundo periodo, siendo estadísticamente significativo para ciprofloxacino (p<0.05). La resistencia a quinolonas fue más elevada en HSH. Haemophilus spp presentó mayor resistencia a ampicilina, ciprofloxacino y tetraciclina en el segundo periodo; siendo significativo para tetraciclina (p<0,05). U. urealyticum presentó elevada resistencia a ciprofloxacino (80,7%) y ofloxacino (32,4%) y baja para doxiciclina (0,8%) y tetraciclina (3,5%)(AU)

Objective. To know the best empirical treatment of urethritis in patients at the City Center of Madrid. Methods. 2.021 urethral exudates were analyzed in men between January 2003-December 2007. In addition to the traditional cultures, it was determined the presence of Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, Trichomonas vaginalis and Herpes simplex. The susceptibility of N.gonorrhoeae and Haemophilus spp was performed by disk diffusion method and U. urealyticum by Mycoplasma IST. Results. The percentage of positive samples was: 30.6%. The most frequently isolated microorganisms were: U. urealyticum 9.9%, N. gonorrhoeae 7.4%, C. trachomatis 5.1% and Haemophilus spp 3.8%. The resistance of N. gonorrhoeae in the first period was: penicillin 11.8%, tetracycline 5.9%, ciprofloxacin 8.8% and presence of betalactamase 11.8%. In the second period: penicillin 9.7%, amoxicillin/clavulanic acid 1.4%, tetracycline 8.3%, ciprofloxacin 23.6% and presence of betalactamase 10.5%. Resistance to ciprofloxacin in non-MSM (men having sex with men) was 20% and in MSM 56.2% (p <0.05). Resistance of Haemophilus spp in the first period was: 38.2% ampicillin, amoxicillin/ clavulanic acid 8.8%, clarithromycin 35.3%, cotrimoxazole 64.7%, cefuroxime 5.9%, ciprofloxacin 8.8%, tetracycline 12.1% and presence of betalactamase 26.5%. In the second period: presence of betalactamase 41.9%, ampicillin 53.1%, amoxicillin/ clavulanic acid 9.4%, cefuroxime 9.4%, clarithromycin 18.7%, tetracycline 34.4%, ciprofloxacin 15.6%, and cotrimoxazole 68.7%. Resistance of U. urealyticum was: ciprofloxacin 80.7%, ofloxacin 32.4%, erythromycin 17.5%, azithromycin 9.6%, tetracycline 3.5% and doxycycline 0.8%. Conclusions. N. gonorrhoeae showed a level of resistance to tetracycline and ciprofloxacin higher in the second period, being significant for ciprofloxacin (p<0.05). Quinolone resistance was higher in MSM. Haemophilus spp showed a level of resistance to ampicillin, ciprofloxacin and tetracycline higher in the second period, being significant for tetracycline (p <0.05). U.urealyticum showed high level of resistance to ciprofloxacin (80.7%) and ofloxacin (32.4%) and low level of resistance to doxycycline (0.8%) and tetracycline (3.5%)(AU)