MicroMundo: experimental project fostering contribution to knowledge on antimicrobial resistance in secondary school.

Antimicrobial resistance (AMR) has become a serious global health threat. Education could play a decisive role, so the scope of innovative educational projects, such as MicroMundo, should be analysed. MicroMundo is a service-learning project for the exploration of microbial biodiversity in soils in search of new antibiotics. In order to evaluate the contribution of MicroMundo to the improvement of knowledge about AMR as well as the optimal age of the participants, an open questionnaire before and after the educational intervention (for secondary school students) and a satisfaction survey (for all participants) were carried out. Programme instruction began in university and continued through secondary school; 14 teachers (phase 1) and 137 students from three educational levels (phase 2) participated. MicroMundo was successfully implemented, showing a statistically significant improvement in knowledge about antibiotics, resistance, health and environmental consequences, and possible measures to reverse the problem, at all educational levels. Furthermore, the satisfaction surveys revealed a growing interest in research and science. Therefore, students of general branches who have not yet opted for a scientific line would be the most suitable to conduct the programme. In conclusion, MicroMundo is recommended as a training method to raise awareness about AMR and promote scientific vocations.

Searching for Antimicrobial-Producing Bacteria from Soils through an Educational Project and Their Evaluation as Potential Biocontrol Agents.

Antimicrobial resistance (AMR) is a serious threat to public health due to the lack of effective drugs to combat infectious diseases, which generates the need to search for new antimicrobial substances. In this study, the potential of soil as a source of antimicrobial-producing bacteria (APB) was investigated and the importance of the connection between education and science was emphasized, using service-learning methodologies. Sixty-one soil samples were collected, and 1220 bacterial isolates were recovered. Eighteen of these isolates showed antimicrobial activity against at least 1 of the 12 indicator bacteria tested (including multidrug-resistant and relevant pathogens). The 18 APB were identified by MALDI-TOF and 6 different genera (Bacillus, Brevibacillus, Lysinobacillus, Peribacillus, Streptomyces, and Advenella) and 10 species were identified. The 18 APB were tested for antifungal activity against four phytopathogenic fungi (Botritis cynerea, Lecanicillium fungicola, Trichoderma harzianum, and Cladobotryum mycophilum). Moreover, the antibiotic susceptibility of APB was tested using the disk-diffusion method as well as their ß-hemolytic activity (important safety criteria for potential future applications). A total of 10 of the 18 APB were able to inhibit at least 50% of indicator bacteria tested, including methicillin-resistant Staphylococcus aureus (MRSA), among others. A total of 4 of the 18 APB (3 Bacillus pumilus and 1 Bacillus altitudinis) showed inhibitory activity against two of the four fungal pathogens tested (B. cinerea and L. fungicola), as well as against 5-7 of the 12 bacterial pathogen indicators; these 4 isolates showed susceptibility to the antibiotics tested and lacked ß-hemolytic activity and were considered promising APB for use as potential biocontrol agents. In addition, one Brevibacillus laterosporus strain had activity against 83% of indicator bacteria tested including Escherichia coli, MRSA and other methicillin-resistant staphylococci, as well as vancomycin-resistant enterococci (but not against fungi). These results show that soil is a source of APB with relevant antibacterial and antifungal activities, and also emphasize the importance of education and science to raise public awareness of the AMR problem and the strategies to control it.

Citizen Contribution for Searching for Alternative Antimicrobial Activity Substances in Soil.

Antimicrobial resistance (AMR) is problematic worldwide, and due to the loss of efficiency of many antibiotics, the pressure to discover alternative antimicrobial molecules has increased. Soil harbors a great biodiversity and biomass of microorganisms, and many antibiotics are produced by soil microbiota. Therefore, soil is a promising reservoir to find new antimicrobial agents. In this respect, novel pedagogical strategies regarding the AMR global crisis have recently been developed in different countries worldwide. Highlighted is the service-learning project "MicroMundo" integrated in a global Citizen Science project called "Tiny Earth". Hence, the present work aimed at determining the antimicrobial activity of soil bacteria, the biodiversity of the selected isolates as putative antimicrobial producers, and their antibiotic resistance profile. Moreover, through the MicroMundo project, we tried to illustrate the relevant link between science and education and the benefits of implementing service-learning methodologies to raise awareness of the AMR problem and to contribute to the search for new alternatives. A total of 16 teachers, 25 university students and 300 secondary school students participated in the search for antimicrobial activity on a collection of 2600 isolates obtained from a total of 130 soil samples analysed. In total, 132 isolates (5% of total tested) were selected as potential antimicrobial producers when two indicator bacteria were used (Escherichia coli and Staphylococcus epidermidis); the most frequent genus among these isolates was Bacillus, followed by Pseudomonas, Paenibacillus and Serratia. The antimicrobial activity (AA) of the 132 potential antimicrobial producers was studied in a second step against 15 indicator bacteria (of six genera and thirteen species, including relevant pathogens). Of the 132 potentially producing bacteria, 32 were selected for further characterization. In this respect, 18 isolates showed low AA, 12 isolates were considered as medium producers, and 2 highly antimicrobial-producing isolates were found (Brevibacillus laterosporus X7262 and Staphylococcus hominis X7276) showing AA against 80% of the 15 indicators tested. Moreover, 48% of the antimicrobial-producing bacteria were susceptible to all antibiotics tested. Due to citizen science, antimicrobial-producing bacteria of great interest have been isolated, managing to raise awareness about the problem of AMR.

Antimicrobial Resistance and Antimicrobial Activity of Staphylococcus lugdunensis Obtained from Two Spanish Hospitals.

Staphylococcus lugdunensis is a coagulase-negative-staphylococci (CoNS) that lately has gained special attention in public health as a human pathogen and also as a bacteriocin-producer bacteria. In this study, we characterized 56 S. lugdunensis isolates recovered from human samples in two Spanish hospitals. Antimicrobial susceptibility testing was performed and antimicrobial resistance and virulence genotypes were determined. Antimicrobial activity (AA) production was evaluated by the spot-on-lawn method against 37 indicator bacteria, including multidrug-resistant (MDR) isolates, and the presence of the lugD gene coding for lugdunin bacteriocin was analyzed by PCR. The antibiotic resistance detected was as follows (% resistance/genes detected): penicillin (44.6%/blaZ), oxacillin (1.8%/mecA on SCCmec-V), erythromycin-clindamycin inducible (7.1%/erm(C), msrA), tetracycline (5.3%/tetK), gentamicin and/or tobramycin (3.6%/ant(4')-Ia, acc(6')-aph(2″)), and fosfomycin (21.4%). A MDR phenotype was detected in 5% of isolates. Twenty-one of the S. lugdunensis isolates showed susceptibility to all 20 antibiotics tested (37.5%). The screening for AA revealed 23 antimicrobial producer (AP) isolates with relevant inhibition against coagulase-positive-staphylococci (CoPS), including both methicillin-susceptible and -resistant S. aureus. The lugD gene was detected in 84% of the 56 S. lugdunensis isolates. All of the AP S. lugdunensis isolates (n = 23) carried the lugD gene and it was also detected in 24 of the non-AP isolates, suggesting different gene expression levels. One of the AP isolates stood out due to its high antimicrobial activity against more than 70% of the indicator bacteria tested, so it will be further characterized at genomic and proteomic level.

Identification of Two Novel EPOR Gene Variants in Primary Familial Polycythemia: Case Report and Literature Review.

Primary familial and congenital polycythemia is a rare disease characterized by an increase in red cell mass that may be due to pathogenic variants in the EPO receptor (EPOR) gene. To date, 33 genetic variants have been reported to be associated. We analyzed the presence of EPOR variants in two patients with polycythemia in whom JAK2 pathogenic variants had been previously discarded. Molecular analysis of the EPOR gene was performed by Sanger sequencing of the coding regions and exon/intron boundaries of exon 8. We performed in vitro culture of erythroid progenitor cells. Segregation studies were done whenever possible. The two patients studied showed hypersensitivity to EPO in in vitro cultures. Analysis of the EPOR gene unveiled two novel pathogenic variants. Genetic testing of asymptomatic relatives could guarantee surveillance and proper management.

Tn1546 structures and multilocus sequence typing of vanA-containing enterococci of animal, human and food origin.

OBJECTIVES: To characterize the Tn1546 structure and to perform the genetic typing of 51 PFGE-unrelated vanA-containing enterococci of different origin (clinical, food and faecal samples of healthy humans and healthy poultry). METHODS: Tn1546 structure was characterized by a PCR primer walking strategy and sequencing. Multilocus sequence typing (MLST) was performed for Enterococcus faecium and Enterococcus faecalis strains, and esp and hyl genes were detected by PCR. RESULTS: Nine different Tn1546 structures were identified in the studied strains. Type I was the most prevalent structure (75%) (identical to GenBank M97297). Two new Tn1546 structures were identified (in three clinical and animal strains), containing two new insertion sequences (ISs; ISEfa11 disrupting vanS and ISEfa10 disrupting orf1). An additional new Tn1546 structure was found in one animal strain, containing ISEf1 interrupting vanY and IS1542 in the orf2-vanR region. A high diversity of sequence types (STs) was detected among clinical (6 ST/18 strains) and non-clinical E. faecium strains (18 ST/24 strains). STs associated with clonal complexes CC17 and CC9 were mainly detected among clinical and non-clinical E. faecium strains, respectively. Seven new STs were identified in non-clinical strains. The esp and hyl genes were only found among clinical E. faecium strains. CONCLUSIONS: A moderate variability in Tn1546 structure has been detected among unrelated vanA-containing enterococci of different origins, showing three new structures including two new ISs. A high diversity of STs was detected among E. faecium strains, especially among non-clinical strains, and new STs have been identified.

Long-term impact of an educational antimicrobial stewardship programme in primary care on infections caused by extended-spectrum ß-lactamase-producing Escherichia coli in the community: an interrupted time-series analysis.

BACKGROUND: There is little evidence on the ecological effect and sustainability of antimicrobial stewardship programmes (ASPs) in primary-care settings. We aimed to determine whether a multimodal, educational ASP would be sustainable in the long-term and reduce the incidence of infections caused by extended-spectrum ß-lactamase-producing Escherichia coli in the community by optimising antibiotic use. METHODS: We did this quasi-experimental intervention study in 214 primary health centres of four primary health-care districts in Andalusia, Spain. Local multidisciplinary teams, comprised of general practitioners, paediatricians, primary-care pharmacists, and epidemiologists, were created in each district and implemented a multimodal, education-based ASP. The core activity of the programme consisted of regular one-to-one educational interviews between a reference interviewing physician and prescribing physicians from each centre on the appropriateness of their most recent (same or preceding day) antibiotic prescriptions based on a structured questionnaire. Appropriate prescribing was defined as compliance of all checklist items with the reference guidelines. An average of five educational interviews were scheduled per prescriber per study year. We did an interrupted time-series analysis to assess the effect of the intervention on quarterly antibiotic use (prescription and collection by the patient) and quality of prescriptions (as defined daily doses per 1000 inhabitants per day) and incidence per 1000 inhabitants of E coli producing extended-spectrum ß-lactamase (ESBL) isolated from urine samples. FINDINGS: The study was done between January, 2012, and December, 2017, in a pre-intervention period of 2012-13 and an intervention period of 2014-17. Throughout the study period, there were 1387 physicians (1116 general practicioners and 271 paediatricians) in the included health centres serving a mean population of 1 937 512 people (299 331 children and 1 638 181 adults). 24 150 educational interviews were done over the 4 years. Inappropriate antibiotic prescribing was identified in 1794 (36·5%) of 4917 educational interviews in 2014 compared with 1793 (26·9%) of 6665 in 2017 (p<0·0001). The intervention was associated with a sustained reduction in the use of ciprofloxacin (relative effect -15·9%, 95% CI -23·9 to -8·0) and cephalosporins (-22·6%, -35·9 to -9·2), and a sustained increase in the use of amoxicillin (22·2%, 6·4 to 38·0) and fosfomycin trometamol (6·1%, 2·6 to 9·6). The incidence density of ESBL-producing E coli decreased by -0·028 cases per 1000 inhabitants (95% CI -0·034 to -0·021) after the start of the programme, reversing the pre-intervention increase and leading to a relative reduction of -65·6% (-68·2 to -63·0) 4 years later. INTERPRETATION: Our data suggest that implementation of a multimodal ASP in primary care that is based on individual educational interviews improves the use of antibiotics and results in a sustained significant reduction of infections by ESBL-producing E coli in the community. This information should encourage the implementation of ASPs in primary care. FUNDING: Instituto de Salud Carlos III, Spanish Government (PI14/01523).

[Suicide Mortality in Andalusia, Spain: geographical Distribution and Relationship with Antidepressants, Altitude and Socioeconomic Inequalities]. / Mortalidad por suicidio en Andalucía: distribución geográfica y relación con el uso de antidepresivos, la altitud y desigualdades socioeconómicas.

BACKGROUND: The suicide mortality rate in Andalusia is higher than the Spanish average, and it shows a different evolution over time. This study analyzes recent changes of suicide mortality in Andalusia, its geographical distribution and its relation with antidepressant use, altitude and socioeconomic inequality. METHODS: An ecological study of the Basic Health Zones in Andalusia has been conducted. Age-standarized suicide rates in 2012 and during 2007-2011 were calculated. The correlation among variables and their association with the geographical differences was assessed with Spearman's coefficient and generalized linear models were also estimated. RESULTS: Andalusia registered 7,58 suicides per 100.000 inhabitants per year from 2007 to 2011, with a range of 0,96 to 27,71 in the different areas. Positive correlation has been observed between altitude and suicide rates in 2012 (r=0,29; p<0,001) and during the period 2007-2011 (r=0,47; p<0,001). Antidepressant use in 2012 was not correlated with suicide rates in that year (r=0,03; p=0,67), but a positive correlation was observed with the precedent five years suicide rates (r=0,18; p=0,008). The Deprivation Index had a positive correlation with suicide rates in 2012 (r=0,29; p<0,001) and in 2007-2011 (r=0,33; p<0,001). CONCLUSIONS: There is a clear geographical pattern in the distribution of suicide mortality in Andalusia and it remains stable over time. Central areas and in those with the highest altitude concentrate the highest suicide rates, which are associated with a larger use of antidepressants and also with higher levels of material deprivation.

[Suicide, antidepressant prescription and unemployment in Andalusia (Spain)]. / Suicidio, prescripción de antidepresivos y desempleo en Andalucía.

OBJECTIVES: To analyze the trend in suicide mortality in Andalusia from 1975 to 2012 and its relationship with unemployment and the use of antidepressants. METHODS: Poisson's segmented regression models were used to estimate changes over time. The association between suicide and the factors examined was measured using Spearman's correlation coefficient. RESULTS: Suicide mortality patterns in men and women are rising. The largest increase was found in people aged from 15 to 44 years, with an annual percentage rate change of 1.21 (95%CI: 0.7-1.7) for men and 0.93 (95%CI: 0.4-1.4) for women. CONCLUSIONS: Mortality by suicide has increased in Andalusia since 1975 in all age and gender groups except for women aged 65 years or above. During the last few decades, an upward trend has been observed in young people and a stable or falling trend in the remaining population. Temporary variations in suicide rates are not associated with unemployment rates or with changes in antidepressant prescription.

Mortalidad por suicidio en Andalucía: distribución geográfica y relación con el uso de antidepresivos, la altitud y desigualdades socioeconómicas / Suicide Mortality in Andalusia, Spain: Geographical Distribution and Relationship with Antidepressants, Altitude and Socioeconomic Inequalities

Fundamentos: Andalucía tiene una mortalidad por suicidio superior a la media española y una evolución temporal discordante con el conjunto del país. El objetivo fue analizar la evolución reciente de la mortalidad por suicidio en Andalucía, su distribución geográfica y la relación con el uso de antidepresivos, la altitud y las desigualdades socioeconómicas. Métodos: Estudio ecológico cuya unidad de análisis fueron las Zonas Básicas de Salud de Andalucía. Se calcularon las tasas de suicidio ajustadas por edad del periodo 2007-2011 y de 2012. Para analizar las variables asociadas a las diferencias geográficas se midió la correlación mediante el coeficiente de Spearman y se estimaron modelos lineales generalizados. Resultados: Andalucía tuvo una tasa de mortalidad por suicidio de 7,58/100.000 habitantes/año entre 2007 y 2011, con un intervalo de 0,96 a 27,71. Hubo correlación positiva entre altitud y tasas de suicidio en 2012 (r=0,29; p<0,001) y en el periodo 2007-2011 (r=0,47; p<0,001). En 2012 el uso de antidepresivos no se correlacionó con las tasas de suicidio (r=0,03; p=0,67) pero sí durante los cinco años anteriores (r=0,18; p=0,008). El Índice de Privación Material tuvo una correlación positiva con las tasas en 2012 (r=0,29; p<0,001) y de 2007-2011 (r=0,33; p<0,001). Conclusiones: Existe un claro patrón geográfico en la distribución de la mortalidad por suicidio en Andalucía que permanece estable en el período estudiado. La zona central y las de mayor altitud concentran las tasas de suicidio más altas y se asocian con utilización de antidepresivos y mayor privación material (AU)

Background: The suicide mortality rate in Andalusia is higher than the Spanish average, and it shows a different evolution over time. This study analyzes recent changes of suicide mortality in Andalusia, its geographical distribution and its relation with antidepressant use, altitude and socioeconomic inequality. Methods: An ecological study of the Basic Health Zones in Andalusia has been conducted. Age-standarized suicide rates in 2012 and during 2007-2011 were calculated. The correlation among variables and their association with the geographical differences was assessed with Spearman’s coefficient and generalized linear models were also estimated. Results: Andalusia registered 7,58 suicides per 100.000 inhabitants per year from 2007 to 2011, with a range of 0,96 to 27,71 in the different areas. Positive correlation has been observed between altitude and suicide rates in 2012 (r=0,29; p<0,001) and during the period 2007-2011 (r=0,47; p<0,001). Antidepressant use in 2012 was not correlated with suicide rates in that year (r=0,03; p=0,67), but a positive correlation was observed with the precedent five years suicide rates (r=0,18; p=0,008). The Deprivation Index had a positive correlation with suicide rates in 2012 (r=0,29; p<0,001) and in 2007-2011 (r=0,33; p<0,001). Conclusions: There is a clear geographical pattern in the distribution of suicide mortality in Andalusia and it remains stable over time. Central areas and in those with the highest altitude concentrate the highest suicide rates, which are associated with a larger use of antidepressants and also with higher levels of material deprivation (AU)

Suicidio, prescripción de antidepresivos y desempleo en Andalucía / Suicide, antidepressant prescription and unemployment in Andalusia (Spain)

Objetivos: Analizar la evolución temporal de la mortalidad por suicidio en Andalucía entre 1975 y 2012, y su relación con el uso de antidepresivos y el desempleo. Métodos: Para estimar los cambios de tendencia se utilizaron modelos segmentados de regresión de Poisson. La correlación se midió mediante el coeficiente de Spearman. Resultados: La tendencia de la mortalidad por suicidio es ascendente, y son las personas de 15 a 44 años de edad las que presentan aumentos más acusados, con un porcentaje de cambio anual de 1,21 (intervalo de confianza del 95% [IC95%]: 0,7-1,7) para los hombres y de 0,93 (IC95%: 0,4-1,4) para las mujeres. Conclusiones: La mortalidad por suicidio ha aumentado desde 1975 en todos los grupos de edad y sexo, salvo en las mujeres ancianas. En las últimas décadas se aprecian tendencias ascendentes en los jóvenes, y estables o descendentes en el resto, que no están asociadas con el desempleo ni con el uso de antidepresivos (AU)

Objectives: To analyze the trend in suicide mortality in Andalusia from 1975 to 2012 and its relationship with unemployment and the use of antidepressants. Methods: Poisson's segmented regression models were used to estimate changes over time. The association between suicide and the factors examined was measured using Spearman's correlation coefficient. Results: Suicide mortality patterns in men and women are rising. The largest increase was found in people aged from 15 to 44 years, with an annual percentage rate change of 1.21 (95%CI: 0.7-1.7) for men and 0.93 (95%CI: 0.4-1.4) for women. Conclusions: Mortality by suicide has increased in Andalusia since 1975 in all age and gender groups except for women aged 65 years or above. During the last few decades, an upward trend has been observed in young people and a stable or falling trend in the remaining population. Temporary variations in suicide rates are not associated with unemployment rates or with changes in antidepressant prescription (AU)

Rapid identification of Enterococcus hirae and Enterococcus durans by PCR and detection of a homologue of the E. hirae mur-2 Gene in E. durans.

During an evaluation of PCR for identification of isolates of Enterococcus hirae, a homologue with 82% identity to E. hirae mur-2 was identified in Enterococcus durans and was named mur-2(ed). PCR using primers for two genes (copY and murG) of E. hirae strains showed amplification with E. hirae strains only. PCR (under high-stringency conditions) with primers for the mur-2(ed) gene gave the expected amplification product only with E. durans strains. A combination of murG and mur-2(ed) primers in a multiplex PCR assay differentiated E. hirae from E. durans in all cases. PCR using these primers appears to be a rapid alternative for identification of E. hirae and E. durans isolates.