Genomic Insights into Staphylococcus aureus Isolates Exhibiting Diminished Daptomycin Susceptibility.

Daptomycin is one of the last therapeutic resources for multidrug-resistant gram-positive bacteria. Despite its structural similarities with glycopeptides, its mechanisms of action and resistance are different and in some respects are not completely understood. Mutations in several genes have been associated with daptomycin resistance, especially in mprF, walkR, rpoB and rpoC, but their role and importance remain to be elucidated. We have studied mutations in 11 genes, which have been previously associated with daptomycin non-susceptibility, in nine daptomycin-non-susceptible Staphylococcus aureus clinical isolates (daptomycin MIC: >1 mg/L). Susceptibility to daptomycin, vancomycin, linezolid, oxacillin, telavancin and dalbavancin was studied. walkR, agrA, cls1, cls2, fakA, pnpA, clpP, prs, rpoB, rpoC and mprF were amplified by PCR and sequenced. The sequences were compared with the S. aureus ATCC 25923 complete genome (GenBank gi: 685631213) by using BLAST® software. We did not find any changes in walkR, pnpA, prs and clpP. All isolates excepting isolate MSa5 showed a high number of significant mutations (between 13 and 25 amino acid changes) in mprF. Most isolates also showed mutations in the rpo genes, the cls genes and fakA. Daptomycin non-susceptibility in S. aureus clinical isolates seems to be reached through different mutation combinations when compared to S. aureus ATCC 25293. Especially mprF and cls1 showed very high polymorphism in most isolates. Meanwhile, one isolate, MSa5, showed only single mutation in mprF (P314T).

GEHEP 010 study: Prevalence and distribution of hepatitis B virus genotypes in Spain (2000-2016).

OBJECTIVE: To study the prevalence and distribution of HBV genotypes in Spain for the period 2000-2016. METHODS: Retrospective study recruiting 2559 patients from 17 hospitals. Distribution of HBV genotypes, as well as sex, age, geographical origin, mode of transmission, HDV-, HIV- and/or HCV-coinfection, and treatment were recorded. RESULTS: 1924 chronically HBV native Spanish patients have been recruited. Median age was 54 years (IQR: 41-62), 69.6% male, 6.3% HIV-coinfected, 3.1% were HCV-coinfected, 1.7% HDV-co/superinfected. Genotype distribution was: 55.9% D, 33.5% A, 5.6% F, 0.8% G, and 1.9% other genotypes (E, B, H and C). HBV genotype A was closely associated with male sex, sexual transmission, and HIV-coinfection. In contrast, HBV genotype D was associated with female sex and vertical transmission. Different patterns of genotype distribution and diversity were found between different geographical regions. In addition, HBV epidemiological patterns are evolving in Spain, mainly because of immigration. Finally, similar overall rates of treatment success across all HBV genotypes were found. CONCLUSIONS: We present here the most recent data on molecular epidemiology of HBV in Spain (GEHEP010 Study). This study confirms that the HBV genotype distribution in Spain varies based on age, sex, origin, HIV-coinfection, geographical regions and epidemiological groups.

Should parasitic disease be investigated in immigrant children with relative eosinophilia from tropical and sub-tropical regions?

BACKGROUND: Immigrants to Spain are mainly from low- and middle-income countries, and around 20% are children. Absolute eosinophilia is defined as >0.45×109 eosinophilic leucocytes/L of peripheral blood. Absolute eosinophilia in travelers and immigrants from tropical and sub-tropical areas is frequently associated with parasitic diseases. However, the significance of relative eosinophilia in immigrant children, defined as >5% eosinophilic leucocytes in those with <0.45×109 eosinophils/L, is unresolved. OBJECTIVES: To describe the importance of relative eosinophilia in a cohort of immigrant children (<18 years) from sub-Saharan Africa, North Africa and Latin America. METHODS: 176 immigrant children without absolute eosinophilia were prospectively evaluated. RESULTS: 25 of them (14.2%) had relative eosinophilia. 10 patients with relative eosinophilia had no diagnosis. 15 with relative eosinophilia (60%) were diagnosed with a parasitic disease, 7 (46.7%) of whom had only one parasite, while co-infection accounted for 8 of the 15 cases (53.3%). Of the parasitic infections, the most frequent causes of relative eosinophilia were filariasis spp. (7/15, 46.7%), strongyloides spp. (5/15, 33.3%), schistosoma spp. (4/15, 26.6%) and Ascaris lumbricoides (2/15, 13.3%). CONCLUSION: The findings suggest that relative eosinophilia is frequently associated with helminthic infection in immigrant children from tropical and sub-tropical areas, so a thorough parasitological study is highly advisable in this group of patients.

Relevance of eosinophilia and hyper-IgE in immigrant children.

Immigrants from undeveloped countries are a growing problem in Europe. Spain has become a frequent destination for immigrants (20% of whom are children) because of its geographic location and its historic and cultural links with Africa and Latin America. Eosinophilia is frequent in adult immigrants, travelers and expatriates coming from tropical areas. However, there are few studies that focus on the incidence and causes of tropical eosinophilia and hyper-IgE in immigrant children.We evaluated, prospectively, the prevalence and causes of eosinophilia and hyper-immunoglobulin E (IgE) in 362 immigrant children coming from Sub-Saharan Africa, Northern Africa and Latin America to Salamanca, Spain, between January 2007 and December 2011.Absolute eosinophilia and hyper-IgE were present in 22.9% and 56.8% of the analyzed children, respectively. The most frequent causes of absolute eosinophilia were filariasis (52.6%), strongyloidiasis (46.8%) and schistosomiasis (28.9%). Filariasis (41.9%), strongyloidiasis (29.6%) and schistosomiasis (22.2%) were the most frequent causes of increased levels of IgE. The area under the ROC curve showed similar values between eosinophil count and IgE levels in the diagnosis of helminthiasis (69% [95% confidence interval (CI) 63%-74%] vs 67% [95% CI 60%-72%], P = 0.24). Eosinophilia and hyper-IgE have a high value as biomarkers of helminthiasis in children coming from tropical and subtropical areas.

Evaluación del citómetro UF-1000i como método de cribado en el diagnóstico de infecciones del tracto urinario / Evaluation of the Sysmex UF-1000i flow cytometer for screening of urinary tract infection

INTRODUCCIÓN: El cultivo de orina supone una enorme carga de trabajo en el Laboratorio de Microbiología y sigue siendo técnica de referencia para el diagnóstico de las infecciones urinarias. Considerando la elevada prevalencia de resultados negativos, la implementación de un método de cribado fiable y rápido podría suponer un ahorro en costes de carga de trabajo y adelantar los resultados negativos. MÉTODO: Evaluamos la utilidad del citómetro de flujo UF-1000i® (bioMérieux, España) para cribado de muestras negativas que se pueden excluir del cultivo. Dividimos las muestras en 2 grupos: grupo 1, hombres y mujeres en edad fértil, que se consideran positivas con un crecimiento ≥ 104 UFC/ml, y grupo 2, consideradas positivas con crecimiento ≥ 105 UFC/ml. RESULTADOS: Enfrentando los datos del cultivo y del cribado en curva ROC, los puntos de mejor sensibilidad y especificidad fueron de 53,1 bacterias/μl para el grupo 1, y de 128,35 bacterias/μl para el grupo 2. En el grupo 1 la sensibilidad fue del 92,2%, la especificidad del 60%, la reducción de cultivos de orina del 46%, con el 2,1% de falsos negativos (42 muestras). En el grupo 2, la sensibilidad fue del 86%, la especificidad del 87,7%, la reducción de cultivos del 57,5%, con el 5,1% de falsos negativos (74 muestras). CONCLUSIÓN: La incorporación del citómetro UF-1000i al cribado de las muestras de orina depende mucho de las características de los pacientes y de la definición de cultivo de orina positivo. En nuestro caso, con el estudio exclusivo de la bacteriuria, los datos de reducción de carga de trabajo y de falsos negativos cuestionan seriamente esta incorporación

INTRODUCTION: The urine culture is a huge workload in the Microbiology Laboratory and remains the gold standard for the diagnosis of urinary tract infections. Considering the high prevalence of negative results, the implementation of a reliable screening method could lead to cost saving in the workload, and speedup reporting of negative results. METHODS: We evaluated the usefulness of the flow cytometer UF-1000i in the screening for negative samples than could be excluded from culture. We divided the samples into two groups, Group 1, males and women of childbearing age who were considered positive with a growth ≥ 104 CFU/ml, and Group 2,considered positive with ≥ 105 CFU/ml growth. RESULTS: On comparing the culture and screening data in the ROC curve, the best sensitivity and specificity points were 53.1 bact/l for Group 1, and 128.3 bact/l for Group 2. In Group 1, the sensitivity was 92.2%and a specificity of 60%, a reduction in urine cultures of 46%, with 2.1% false negative (42 samples). In Group 2, the sensitivity was 86%, with a specificity of 87.7%, a culture reduction of 57.5%, and 5.1% false negatives (74 samples). CONCLUSION: The incorporating of the UF-1000i cytometer to the screening of urine samples depends on the characteristics of the patients and the definition of positive urine culture. In our case, with only studying bacteriuria, the data on the reduction of workload and the false negatives seriously question this incorporation

[Evaluation of the Sysmex UF-1000i flow cytometer for screening of urinary tract infection]. / Evaluación del citómetro UF-1000i como método de cribado en el diagnóstico de infecciones del tracto urinario.

INTRODUCTION: The urine culture is a huge workload in the Microbiology Laboratory and remains the gold standard for the diagnosis of urinary tract infections. Considering the high prevalence of negative results, the implementation of a reliable screening method could lead to cost saving in the workload, and speed up reporting of negative results. METHODS: We evaluated the usefulness of the flow cytometer UF-1000i in the screening for negative samples than could be excluded from culture. We divided the samples into two groups, Group 1, males and women of childbearing age who were considered positive with a growth ≥ 104 CFU/ml, and Group 2, considered positive with ≥ 105 CFU/ml growth. RESULTS: On comparing the culture and screening data in the ROC curve, the best sensitivity and specificity points were 53.1 bact/µl for Group 1, and 128.3 bact/µl for Group 2. In Group 1, the sensitivity was 92.2% and a specificity of 60%, a reduction in urine cultures of 46%, with 2.1% false negative (42 samples). In Group 2, the sensitivity was 86%, with a specificity of 87.7%, a culture reduction of 57.5%, and 5.1% false negatives (74 samples). CONCLUSION: The incorporating of the UF-1000i cytometer to the screening of urine samples depends on the characteristics of the patients and the definition of positive urine culture. In our case, with only studying bacteriuria, the data on the reduction of workload and the false negatives seriously question this incorporation.

Hepatitis B reactivation in a hepatitis B surface antigen-negative patient after allogeneic bone marrow transplant: successful treatment with lamivudine and seroconversion.

Hepatitis B reactivation in hepatitis B surface antigen (HBsAg)-negative and anti-HBsAg antibodies-positive patients is an infrequent complication of chemotherapy, usually with fatal evolution. Here we report an HBsAg-negative patient with a myelodysplastic syndrome, who developed hepatitis B reactivation after chemotherapy and evolved favorably after lamivudine treatment, allowing seroconversion.