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Objectives: Recently a first K. pneumoniae resistant to ertapenem and meropenem was isolated in Dubrava University Hospital, Zagreb, Ceroatia. The aim of the study was to explain the mechanisms of reduced susceptibility to carbapenems in ESBL producing K. pneumoniae that occured during therapy. Methods: Initially an ESBL producing K.pneumoniae was isolated from blood culture of the cardiac transplant patient. The patient was treated with meropenem twice in six weeks. Two weaks later K. pneumoniae with reduced suscepeptibility to carbapenems was isolated from anal swab. ESBL was detected by DDST. MIC was determined by broth microdilution according to CLSI. Modified Hodge Test(MHT) was used to screen for production of carbapenemases. MBL E-test was used to screen for production of metallo-beta-lactamases. The presence of blaSHV, blaTEM, blaCTX-M, blaACT-1, blaKPC, blaOXA-23, blaOXA-51, blaOXA-58, blaOXA-69 and blaVIM was determined by PCR. The genetic relatedness of the strains was investigated using PFGE. To study porin content of the strains, outer membrane proteins (OMPs) were extracted and separated by SDS-PAGE. Results: The initial isolate was susceptible to ertapenem (MIC 0.125 mg/L), meropenem (MIC 0.032 mg/l) and imipenem (MIC 0.25mg/l). The strain obtained after patient was treated with meropenem showed reduced carbapenem susceptibility: ertapenem MIC 32mg/l , meropenem MIC 16 mg/l, and imipenem MIC 8 mg/l. PFGE profile showed that strains are highly related. Both strains were shown to possess blaSHV and blaCTX-M genes by PCR. Sequencing of bla ESBL genes revealed the presence of CTX-M-15/28 and SHV-11 beta lactamases in both strains. MHT was negative in both strains. The PCR reactions with primers specific for ACT-1, KPC and oxacillinases were negative. Both isolates produced OMP-A like protein. Initial isolate expressed one single porin (Omp36) while second isolate did not produce any of the two major porins of K. pneumoniae (Omp35 or Omp36). Conclusion: K. pneumoniae with decreased carbapenem susceptibility was isolated from surveillance culture (anal swab) after prolonged meropenem therapy. Since no carbapenemases were produced by the strains it is likely to expect that carbapenem resistance was due to hyperproduction of CTX-M beta–lactamase combined with porin loss. This study highlights the need to establish an antimicrobial resistance survellance network for K.pneumoniae and to further monitor the trends and new

The increasing prevalence of nosocomial infections produced by multidrugresistant (MDR) or extensively drug-resistant (XDR) Pseudomonas aeruginosa is frequently linked to widespread international strains designated high-risk clones. In this work, we attempted to decipher the interplay between resistance profiles, high-risk clones, and virulence, testing a large (n 140) collection of well-characterized P. aeruginosa isolates from different sources (bloodstream infections, nosocomial outbreaks, cystic fibrosis, and the environment) in a Caenorhabditis elegans infection model. Consistent with previous data, we documented a clear inverse correlation between antimicrobial resistance and virulence in the C. elegans model. Indeed, the lowest virulence was linked to XDR profiles, which were typically linked to defined high-risk clones. However, virulence varied broadly depending on the involved highrisk clone; it was high for sequence type 111 (ST111) and ST235 but very low for ST175. The highest virulence of ST235 could be attributed to its exoU type III secretion system (TTSS) genotype, which was found to be linked with higher virulence in our C. elegans model. Other markers, such as motility or pigment production, were not essential for virulence in the C. elegans model but seemed to be related with the higher values of the statistical normalized data. In contrast to ST235, the ST175 high-risk clone, which is widespread in Spain and France, seems to be associated with a particularly low virulence in the C. elegans model. Moreover, the previously described G154R AmpR mutation, prevalent in ST175, was found to contribute to the reduced virulence, although it was not the only factor involved. Altogether, our results provide a major step forward for understanding the interplay between P. aeruginosa resistance profiles, high-risk clones, and virulence., This work was supported by the Planes Nacionales de IDi 2008-2011/2013-2016 and Instituto de Salud Carlos III, Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía y Competitividad, Spanish Network for Research in Infectious Diseases (REIPI RD12/0015/0006, RD12/0015/004, RD16/0016/ 0004, RD16/0016/008, and RD16/0016/0011) and grants PI15/00088 and PI12/00734. This work was cofinanced by grants from the European Development Regional Fund “A way to achieve Europe” and operative program Intelligent Growth (2014-2020). I.S.-D. is the recipient of a P-FIS fellowship from Instituto de Salud Carlos III, Ministerio de Economía y Competitividad.

This study assessed the molecular epidemiology, resistance mechanisms, and susceptibility profiles of a collection of 150 extensively drug-resistant (XDR) Pseudomonas aeruginosa clinical isolates obtained from a 2015 Spanish multicenter study, with a particular focus on resistome analysis in relation to ceftolozane-tazobactam susceptibility. Broth microdilution MICs revealed that nearly all (>95%) of the isolates were nonsusceptible to piperacillin-tazobactam, ceftazidime, cefepime, aztreonam, imipenem, meropenem, and ciprofloxacin. Most of them were also resistant to tobramycin (77%), whereas nonsusceptibility rates were lower for ceftolozane-tazobactam (31%), amikacin (7%), and colistin (2%). Pulsed-field gel electrophoresis–multilocus sequence typing (PFGE-MLST) analysis revealed that nearly all of the isolates belonged to previously described high-risk clones. Sequence type 175 (ST175) was detected in all 9 participating hospitals and accounted for 68% ( n = 101) of the XDR isolates, distantly followed by ST244 ( n = 16), ST253 ( n = 12), ST235 ( n = 8), and ST111 ( n = 2), which were detected only in 1 to 2 hospitals. Through phenotypic and molecular methods, the presence of horizontally acquired carbapenemases was detected in 21% of the isolates, mostly VIM (17%) and GES enzymes (4%). At least two representative isolates from each clone and hospital ( n = 44) were fully sequenced on an Illumina MiSeq. Classical mutational mechanisms, such as those leading to the overexpression of the β-lactamase AmpC or efflux pumps, OprD inactivation, and/or quinolone resistance-determining regions (QRDR) mutations, were confirmed in most isolates and correlated well with the resistance phenotypes in the absence of horizontally acquired determinants. Ceftolozane-tazobactam resistance was not detected in carbapenemase-negative isolates, in agreement with sequencing data showing the absence of ampC mutations. The unique set of mutations responsible for the XDR phenotype of ST175 clone documented 7 years earlier were found to be conserved, denoting the long-term persistence of this specific XDR lineage in Spanish hospitals. Finally, other potentially relevant mutations were evidenced, including those in penicillin-binding protein 3 (PBP3), which is involved in β-lactam (including ceftolozane-tazobactam) resistance, and FusA1, which is linked to aminoglycoside resistance.

Ante la escasa apropiación por parte de los estudiantes de primaria del énfasis agropecuario de la Institución Educativa Técnica Agropecuaria San Cayetano de Gallego surge la necesidad de desarrollar un material educativo multimedia para introducir presaberes referentes a la línea agrícola del énfasis de la institución en estudiantes de grado quinto. Para lograr lo anterior, se realizó un diagnostico referente al nivel de conocimiento que tienen los estudiantes del tema de horticultura de la línea de trabajo agrícola del énfasis de la institución, así mismo, se realizó el diseño del material educativo multimedia a partir de los criterios metodológicos ADDIE. Y se validó, con docentes de grado quinto y sexto, el material educativo multimedia desarrollado, en referencia a aspectos técnicos, fomento de presaberes de la línea agrícola, operatividad y estructura. La presente investigación pertenece a un enfoque cualitativo puesto que el propósito es examinar la forma en que los individuos perciben y experimentan los fenómenos que los rodean, profundizando en sus puntos de vista, interpretaciones y significados. Por otra parte, el diseño o abordaje general que utiliza es de tipo descriptivo, pues describe o puntualiza en la situación relacionada con la apropiación del énfasis por parte de los estudiantes de quinto grado y su proceso de transición entre la básica primaria y la básica secundaria en lo relacionado con el conocimiento de las temáticas abordadas en cada una de las asignaturas que componen el área técnica. El material multimedia elaborado contiene información de desarrollo teórico, actividades de aprendizajes y actividades de contextualización que propicien el aprendizaje y desarrollo de competencias y habilidades del énfasis agropecuario de la institución propias de la línea agrícola, específicamente de la temática relacionada con la horticultura y de esta manera los estudiantes reconozcan todas las generalidades que se dan para el establecimiento de una huerta familiar o casera y escolar. Given the limited appropriation by primary school students of the agricultural emphasis of the San Cayetano de Gallego Agricultural Technical Educational Institution, the need arises to develop multimedia educational material to introduce pre-knowledge regarding the agricultural line of emphasis of the institution in undergraduate students. fifth. To achieve the above, a diagnosis was made regarding the level of knowledge that students have on the subject of horticulture in the line of agricultural work of the institution's emphasis, likewise, the design of multimedia educational material was carried out based on the criteria ADDIE methodological And the developed multimedia educational material was validated, with fifth and sixth grade teachers, in reference to technical aspects, promotion of pre-knowledge of the agricultural line, operability and structure. The present investigation belongs to a qualitative approach since the purpose is to examine the way in which individuals perceive and experience the phenomena that surround them, delving into their points of view, interpretations and meanings. On the other hand, the design or general approach that it uses is of a descriptive type, since it describes or points out the situation related to the appropriation of emphasis by fifth grade students and their transition process between basic primary and basic secondary in relation to the knowledge of the topics addressed in each of the subjects that make up the technical area. The multimedia material produced contains information on theoretical development, learning activities and contextualization activities that promote learning and development of skills and abilities of the institution's agricultural emphasis, specific to the agricultural line, specifically on the subject related to horticulture and this way the students recognize all the generalities that are given for the establishment of a family or home and school garden. Keywords: multimedia educational material, agriculture, horticulture, school garden, home garden, harmonious transition, pre-knowledge. Licenciado en Informática Educativa http://www.ustadistancia.edu.co/?page_id=3956 Pregrado