Optimization of a viability PCR method for the detection of Listeria monocytogenes in food samples

Postprint (author's final draft

GeNIUL, experts on viability PCR

The standard PCR technique allows the rapid detection of microorganisms, difficult to detect by traditional culture methods. However, the control of samples through PCR has the drawback of false positives due to amplification of DNA/RNA of non-viable cells. This can be overcome by adding intercalating agents (EMA or PMA) to the sample, which selectively penetrates nonviable cells and blocks their DNA/RNA after a photo-activation process, preventing its subsequent amplification by PCR. This process is known as viability PCR (v-PCR). v-PCR has been used as an accurate, reliable and simple method to detect and quantify viable microorganism involved in environmental samples, food production process and clinical samples. GenIUL provides the reagents (EMA and PMA) and the PhAST Blue photo-activation system needed to carry out v-PCR, ensuring optimum and uniform photochemical reaction of intercalating agents prior to PCRPeer Reviewe

Visible light enhances the antimicrobial effect of some essential oils

The photodisinfection is a topical, broad spectrum antimicrobial technology, targeting bacteria, virus, fungi, and protozoa effective for single cells as for biofilms. Natural molecules have been studied less than synthetic agents in the process but they are currently receiving great interest. Therefore, the aim of this study is to evaluate for the first time if non-coherent blue and red light enhances the antimicrobial activity of some essential oils when standard strains for antibiotic or fungicide tests are enlightened in vitro. Staphylococcus epidermidis, Pseudomonas aeruginosa and Candida albicans collection strains were irradiated with monochromatic visible light from light emitting diodes in the presence of 5% and 0.5% eucalyptus (Eucalyptus globulus), clove (Eugenia caryophyllata), and thyme (Thymus vulgaris) essential oils. Microbial levels were measured by plate count on culture media. In this preliminary report, the results differ according to the kind and concentration of antimicrobial oils, the wavelength of light, and the prokaryotic or eukaryotic microorganism. The results support the idea that mainly blue light enhances the innate antimicrobial activity of the essential oils, especially phenols, and could offer a very efficient and natural way to combat microorganisms in several industries and medical applications.Postprint (author's final draft

Presence of Waddlia chondrophila in hot water systems from non-domestic buildings in France

Postprint (author's final draft

Utilidad del Acetato de Medroxiprogesterona en fase folicular para la supresión hipofisaria en donantes de óvulos

El objetivo de esta tesis es evaluar la eficacia del acetato de medroxiprogesterona en fase folicular como supresor hipofisario en la estimulación ovárica. El interés sobre el acetato de medroxiprogesterona radica en las ventajas derivadas de su uso como supresor hipofisario en los protocolos de estimulación ovárica. Estos protocolos son más cómodos y de menor coste que los protocolos convencionales con análogos de la GnRH, y presentan resultados similares en términos de luteinización precoz, número de ovocitos, número de embriones, tasa de implantación, tasa de gestación y aparición de malformaciones congénitas según estudios previamente publicados. Para ello se ha realizado un estudio de no inferioridad con el objetivo de comparar el acetato de medroxiprogesterona frente al ganirelix (antagonista de la GnRH) en donantes de óvulos con el objetivo de prevenir el pico prematuro de LH, y de esta forma obtener mayor respuesta y mayor sincronía en la estimulación ovárica. Se trata de un ensayo clínico de fase III, de no inferioridad, prospectivo, aleatorizado, controlado abierto, de grupos paralelos, llevado a cabo en IVI VALENCIA entre octubre de 2017 y junio de 2019. Se incluyeron 318 donantes con función ovárica normal y fueron asignadas al azar en dos grupos: 161 grupo AMP y 157 grupo ganirelix. La estimulación ovárica se llevó a cabo con 150-225 UI/ día de FSHr. No se encontraron diferencias estadísticamente significativas entre ambos grupos en cuanto a número de ovocitos recuperados, dosis total de gonadotropinas, duración de la estimulación, ni en el perfil endocrino en suero ni en líquido líquido folicular. No se observó ningún caso de ovulación temprana en ninguno de los grupos. No se encontraron diferencias estadísticamente significativas entre ambos grupos con respecto a la tasa de implantación, tasa de embarazo clínico, tasa de embarazo en curso, tasa de nacidos vivos, o tasa acumulativa de nacidos vivos. Como conclusión, nuestro estudio confirma que el acetato de medroxiprogesterona se puede utilizar con éxito como adyuvante de FSHr durante la estimulación ovárica en los ciclos de donantes de ovocitos desencadenados con un agonista de GnRH, ya que permite obtener similar número de ovocitos MII y similares resultados reproductivos con respecto a ganirelix. Nuestros datos no sugieren asociación con un impacto negativo sobre los ovocitos y reivindican un enfoque más amigable para el paciente.The objective of this thesis is to evaluate the efficacy of medroxyprogesterone acetate in the follicular phase as a pituitary suppressor in ovarian stimulation.The interest in medroxyprogesterone acetate lies in the advantages derived from its use as a pituitary suppressor in ovarian stimulation protocols. These protocols are more comfortable and cheaper than conventional protocols with GnRH analogues, and present similar results in terms of early luteinization, number of oocytes, number of embryos, implantation rate, gestation rate and appearance of congenital malformations according to previously published studies. To this end, a non-inferiority study has been carried out with the aim of comparing medroxyprogesterone acetate against ganirelix (GnRH antagonist) in egg donors with the aim of preventing premature peak of LH, and thus obtain greater response and greater synchrony in ovarian stimulation. This is a phase III clinical trial, non-inferiority, prospective, randomized, open controlled, parallel group, carried out at IVI VALENCIA between October 2017 and June 2019. We included 318 donors with normal ovarian function and were randomly assigned into two groups: 161 AMP group and 157 ganirelix group. Ovarian stimulation was carried out with 150-225 IU/day of FSHr. No statistically significant differences were found between the two groups in terms of number of oocytes recovered, total dose of gonadotropins, duration of stimulation, either in the serum endocrine profile or in follicular fluid. No cases of early ovulation were observed in either group. No statistically significant differences were found between the two groups with respect to implantation rate, clinical pregnancy rate, ongoing pregnancy rate, live birth rate, or cumulative live birth rate. In conclusion, our study confirms that medroxyprogesterone acetate can be successfully used as an adjuvant to FSHr during ovarian stimulation in oocyte donor cycles triggered with a GnRH agonist, as it allows to obtain similar number of MII oocytes and similar reproductive results with respect to ganirelix. Our data do not suggest association with a negative impact on oocytes and claim a more patient-friendly approach

Mice with Pulmonary Tuberculosis Treated with Mycobacterium vaccae Develop Strikingly Enhanced Recall Gamma Interferon Responses to M. vaccae Cell Wall Skeleton

Whole heat-killed Mycobacterium vaccae is used as an immunotherapeutic agent in tuberculosis (TB), but the compound(s) that triggers its immunostimulatory ability is not known. Here, we show that among different subcellular fractions, the cell wall skeleton induced a prominent expression of gamma interferon in splenocytes from both non-TB and TB M. vaccae-treated mice

Avaluació de l'autoestima dels estudiants d'ESO. Estudi de cas en un IES

L'article presenta els resultats de la recerca feta en un institut d'Educació Secundària sobre l'autoestima dels estudiants d'Educació Secundària Obligatòria. Després de definir el concepte d'autoestima i fer un recorregut per les investigacions dutes a terme sobre el tema, s'exposa com les autores han estudiat la relació entre l'autoestima i un seguit de variables personals i també amb el rendiment escolar. L'estudi conclou que l'autoestima influeix en el desenvolupament de l'adolescent tant acadèmic com de personalitat, fins al punt que el grau d'influència és diferent segons el tipus d'autoestima.El artículo presenta los resultados de una investigación llevada a cabo en un instituto de Educación Secundaria sobre la autoestima de los estudiantes de Educación Secundaria Obligatoria. Tras definir el concepto de autoestima y realizar un recorrido por las investigaciones realizadas en torno a la temática, se expone cómo las autoras han estudiado la relación entre autoestima y un séquito de variables personales, y, del mismo modo, con el rendimiento escolar. El estudio concluye que la autoestima influye en el desarrollo del adolescente tanto académica como personalmente, hasta el punto que el grado de influencia es diferente según el tipo de autoestima.This paper shows the outcomes of a research carried out in a secondary school. The research investigates issues related to self-esteem in High School students. After defining the concept of self-esteem and revising other researches on the subject matter, we show how we have studied the relationship between self-esteem, personal variables and school performance. The study comes to the conclusion that self-esteem has an influence on academic and personal development during adolescence, being the degree of influence different depending on the type of self-esteem

Amoeba-related health risk in drinking water systems: could monitoring of amoebae be a complementary approach to current quality control strategies?

Postprint (published version

Viability qPCR, a new tool for Legionella risk management

Background Viability quantitative Polymerase Chain Reaction (v-qPCR) is a recent analytical approach for only detecting live microorganisms by DNA amplification-based methods This approach is based on the use of a reagent that irreversibly fixes dead cells DNA. In this study, we evaluate the utility of v-qPCR versus culture method for Legionellosis risk management. Methods The present study was performed using 116 real samples. Water samples were simultaneously analysed by culture, v-qPCR and qPCR methods. Results were compared by means of a non-parametric test. Results In 11.6% of samples using both methods (culture method and v-qPCR) results were positive, in 50.0% of samples both methods gave rise to negative results. As expected, equivalence between methods was not observed in all cases, as in 32.1% of samples positive results were obtained by v-qPCR and all of them gave rise to negative results by culture. Only in 6.3% of samples, with very low Legionella levels, was culture positive and v-qPCR negative. In 3.5% of samples, overgrowth of other bacteria did not allow performing the culture. When comparing both methods, significant differences between culture and v-qPCR were in the samples belonging to the cooling towers-evaporative condensers group. The v-qPCR method detected greater presence and obtained higher concentrations of Legionella spp. (p < 0.001). Otherwise, no significant differences between methods were found in the rest of the groups. Conclusions The v-qPCR method can be used as a quick tool to evaluate Legionellosis risk, especially in cooling towers-evaporative condensers, where this technique can detect higher levels than culture. The combined interpretation of PCR results along with the ratio of live cells is proposed as a tool for understanding the sample context and estimating the Legionellosis risk potential according to 4 levels of hierarchy