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

Presence of Listeria monocytogenes in prepared foods. Analysis of influencing factors

Although food-borne outbreaks of listeriosis are uncommon, they remain a major public health problem. We assessed the prevalence of L. monocytogenes in prepared foods and the influence of different factors (including type of food or presence of accompanying bacteria). Results showed that accompanying bacteria cause interference in the sensitivity of the detection method, being half Fraser the enrichment medium of choice. In 2760 samples, the global prevalence of L. monocytogenes was 1.4%. In ready-to-eat foods, the highest prevalence (23.3%) was found in products containing fermented or cured ingredients and the prevalence was also higher in foods with high counts of aerobic bacteria and lactose positive Enterobacteriaceae, two indicators of process hygiene. We also found that foods stored <0ºC had a higher prevalence (4.1%) of L. monocytogenes. In prepared foods, factors favouring the presence of L. monocytogenes could be some components of ready-to-eat foods, temperatures <0ºC and presence of accompanying bacteria

Pasaporte a la profesión en el Grado de Farmacia

La incorporació del sistema universitari espanyol al Espai Europeu d’Educació Superior (EEES) ha implicat una sèrie d’adaptacions dels graus. Entre aquests s’ha de destacar un període de pràctiques externes a la pròpia universitat, lo que ha comportar una sèrie de canvis considerables en els plans d’estudis de manera que s’aconsegueixi un balanç més equilibrat entre la formació teòrica i pràctica rebuda pels estudiants. Aquest nou marc de treball ha estat confirmat legalment pel Govern espanyol (Real Decreto 1393/2007, de 20 d’octubre i Real Decreto 1707/2011 de 18 de novembre) i en el cas de la Universitat de Barcelona amb la publicació d’una normativa de pràctiques (Normativa de pràctiques acadèmiques externes dels estudiants de la Universitat de Barcelona). Aprofitant aquesta oportunitat per adaptar el Grau de Farmàcia a la realitat social actual, l’equip deganal de la Facultat de Farmàcia amb el recolzament del personal administratiu i el Servei d’atenció a l’estudiant, va assumir el repte d’incloure una nova assignatura en el pla docent del Grau de Farmàcia, que es va denominar “Pràctiques en empreses”. En paral·lel a la posta en marxa d’aquesta assignatura, es va iniciar una nova activitat per assegurar que els estudiants escollirien adequadament la companyia/departament/lloc de treball i donar-los les nocions bàsiques per poder afrontar amb les millors garanties d’èxit l’entrevista laboral. Sota el nom “Passaport a la professió”, s’han programat una sèrie de deu sessions per cada any acadèmic. Aquestes sessions inclouen temes d’una amplia varietat per proveir als estudiants amb les eines bàsiques per aprofitar al màxim el període de practiques i afrontar amb èxit el seu futur professional quan acabin els estudis de grau. A més, s’han celebrat tres workshops i dos taules rodones per apropar el mon de l’empresa a la universitat. S’ha de destacar que el projecte s’ha ampliat a nivell internacional, ja que una empresa farmacèutica amb seu en el Regne Unit cada any contracta a dos o tres estudiants de grau durant un any. Les dades estadístiques obtingudes en el procés s’han analitzat per tenir una millor comprensió de l’activitat i poder millorar el programa

GVPC Medium Manufactured without Oxygen Improves the Growth of Legionella spp. and Exhibits Enhanced Selectivity Properties

ABSTRACTGlycine-vancomycin-polymyxin-cycloheximide agar (GVPC) is a recommended medium for the detection of Legionella spp. in water samples. However, its quality could be improved in terms of recovery of Legionella spp. and selectivity properties. Modifications were introduced in GVPC manufacture: autoclaving conditions (115°C, 15 min) and atmosphere during component-stirring (removal of oxygen and N2 injection). The use of softer autoclaving conditions (115°C, 15 min) improved the growth of Legionella anisa by the spiral method and Legionella pneumophila after membrane filtration. The medium manufactured with O2 removal and autoclaving for 15 min at 115°C allowed a faster growth of L. pneumophila (colonies visible at day 2) and a notable increase of L. anisa growth (colonies appearing at day 3, and statistically significant numbers of CFU at day 5). After 3 to 5 days of incubation, the improved media showed higher selectivity properties, particularly for Enterococcus faecalis ATCC 29212 and Pseudomonas aeruginosa ATCC 9027. A further improvement was achieved by the addition of N2 during ingredient stirring, leading to a statistically significant faster growth of L. pneumophila at days 2 and 3 and L. anisa at day 3. Selectivity properties were also enhanced, resulting in the complete inhibition of both E. faecalis strains and Escherichia coli and complete-partial inhibition of P. aeruginosa. Oxygen removal during GVPC manufacture using a vacuum pump system promotes the growth of L. pneumophila and L. anisa, and markedly inhibits the growth of E. coli, P. aeruginosa, and E. faecalis. IMPORTANCE Currently, GVPC is a recommended medium for the detection of Legionella spp. in water samples. However, recovery of Legionella spp. and selectivity properties can be improved. GVPC medium manufactured without oxygen improved the growth of Legionella pneumophila and Legionella anisa. Oxygen removal during GVPC manufacture also improved selectivity properties. A further improvement was achieved by the addition of N2 during ingredient stirring, leading to a faster growth of L. pneumophila at days 2 and 3 and L. anisa at day 3 and enhancement of selectivity properties. The introduction of the modified GVPC medium in routine practice can allow a better detection of Legionella spp. in water samples

Dopamine genes (DRD2/ANKK1-TaqA1 and DRD4-7R) and executive function: their interaction with obesity

Obesity is a multifactorial disease caused by the interaction between genotype and environment, and it is considered to be a type of addictive alteration. The A1 allele of the DRD2/ANKK1-TaqIA gene has been associated with addictive disorders, with obesity and with the performance in executive functions. The 7 repeat allele of the DRD4 gene has likewise been associated with the performance in executive functions, as well as with addictive behaviors and impulsivity. Participants were included in the obesity group (N = 42) if their body mass index (BMI) was equal to or above 30, and in the lean group (N = 42) if their BMI was below 25. The DRD2/ANKK1-TaqIA and DRD4 VNTR polymorphisms were obtained. All subjects underwent neuropsychological assessment. Eating behavior traits were evaluated. The 'DRD2/ANKK1-TaqIA A1-allele status' had a significant effect on almost all the executive variables, but no significant 'DRD4 7R-allele status' effects were observed for any of the executive variables analyzed. There was a significant 'group' x 'DRD2/ANKK1-TaqIA A1-allele status' interaction effect on LN and 'group' x 'DRD4 7R-allele status' interaction effect on TMT B-A score. Being obese and a carrier of the A1 allele of DRD2/ANKK1-TaqIA or the 7R allele of DRD4 VNTR polymorphisms could confer a weakness as regards the performance of executive functions

Salmosan, a β-Galactomannan-Rich Product, Protects Epithelial Barrier Function in Caco-2 Cells Infected by Salmonella enterica Serovar Enteritidis

Background: One promising strategy for reducing human salmonellosis induced by Salmonella Enteritidis is to supplement animal diets with natural feed additives such as mannan oligosaccharides (MOS). Objective: The aim of this study was to investigate the potential role of Salmosan® (S-βGM), an extremely β-galactomannan-rich MOS product, in preventing epithelial barrier function disruption induced by Salmonella Enteritidis colonization in an in vitro model of intestinal Caco-2 cells in culture. Methods: Differentiated Caco-2 cells were incubated for 3 h with Salmonella Enteritidis at a multiplicity of infection 10 (MOI 10) in the absence or presence of 500 µg/mL S-βGM. Paracellular permeability (PP) was assessed by transepithelial electrical resistance (TER), D-mannitol and fluorescein isothiocyanate-dextran (FD-4) flux. Tight junction (TJ) proteins and cytoskeletal actin were also localized by confocal microscopy. Reactive oxygen species (ROS) and lipid peroxidation products were evaluated. Scanning and transmission electron microscopy were used to visualize Salmonella Enteritidis adhesion to, and invasion of, the Caco-2 cell cultures. Results: Compared to controls, TER was significantly reduced 30 %, and D-mannitol and FD-4 flux were significantly increased 374 % and 54 % in Salmonella Enteritidis-infected cultures. The presence of S-βGM in infected cultures induced total recoveries of TER and FD-4 flux to values that did not differ from control (P = 0.07 and P = 0.55, respectively), and a partial recovery of D-mannitol flux. These effects were confirmed by immunolocalization of actin, zonula occludens protein-1 (ZO-1) and occludin. Similar results were obtained for Salmonella Dublin. The protection of S-βGM on PP in infected cultures may be associated with a total recovery of ROS production to values that did not differ from control (P = 0.11). Moreover, S-βGM has the capacity to agglutinate bacteria, leading to a significant reduction in intracellular Salmonella Enteritidis of 32 % (P < 0.05). Conclusions: The results demonstrate that S-βGM contributes to protecting epithelial barrier function in a Caco-2 cell model disrupted by Salmonella Enteritidis

Improvement of Mueller-Kauffman Tetrathionate-Novobiocin (MKTTn) enrichment medium for the detection of Salmonella enterica by the addition of wx situ-generated tetrathionate

The detection of Salmonella in food is based on the use of a selective enrichment broth such as Muller-Kauffman Tetrathionate-Novobiocin (MKTTn), in which tetrathionate plays a key role by providing Salmonella with a growth advantage. As sodium tetrathionate is unstable, it is generated in situ by the addition of iodine (Lugol's solution) before seeding. This step is cumbersome as the solution is easily spilled, compromising the performance of the medium and hindering the work of technicians. The aim of this study was to optimize MKTTn broth by generating tetrathionate ex situ through an external reaction between iodine and thiosulphate followed by lyophilization. Quality control procedures were performed to compare the modified and original media, testing pure productivity (enrichment with 50-120 CFU of Salmonella Thyphimurim ATCC 14028 and Salmonella Enteritidis ATCC 13076 and plating on Xylose Lysine Deoxycholate agar, XLD), mixed productivity (50-120 CFU of Salmonella strains and Pseudomonas aeruginosa and Escherichia coli at ≥104 CFU and XLD plating) and selectivity (≥104 CFU of P. aeruginosa and Enterococcus faecalis and plating on Tryptone Casein Soy agar, TSA). The modified MKTTn medium (S/L) performed comparably with the original medium in terms of growth of both Salmonella strains (>300 colonies in XLD), alone or with P. aeruginosa and E. coli. Quantitative assays showed no statistically significant differences in the number of colonies grown on XLD after 10-5 dilution (p=0.7015 with S. Thyphimurim ATCC 14028 and p=0.2387 with S. Enteritidis ATCC 13076; ANOVA test). MKTTn medium (S/L) was also selective against E. coli (≤100 colonies) and E. faecalis (<10 colonies). These results suggest that adding tetrathionate as a lyophilisate (S/L) is a feasible alternative to the use of Lugol's solution for the preparation of MKTTn enrichment broth and does not affect the properties of the mediu

Arbovirus surveillance: first dengue virus detection in local Aedes albopictus mosquitoes in Europe, Catalonia, Spain, 2015

Dengue has emerged as the most important viral mosquito-borne disease globally. The current risk of dengue outbreaks in Europe appeared with the introduction of the vector Aedes albopictus mosquito in Mediterranean countries. Considering the increasing frequency of dengue epidemics worldwide and the movement of viraemic hosts, it is expected that new autochthonous cases will occur in the future in Europe. Arbovirus surveillance started in Catalonia in 2015 to monitor imported cases and detect possible local arboviral transmission. During 2015, 131 patients with a recent travel history to endemic countries were tested for dengue virus (DENV) and 65 dengue cases were detected. Twenty-eight patients with a febrile illness were viraemic, as demonstrated by a positive real-time RT-PCR test for DENV in serum samples. Entomological investigations around the viraemic cases led to the detection of DENV in a pool of local Ae. albopictus captured in the residency of one case. The sequence of the DENV envelope gene detected in the mosquito pool was identical to that detected in the patient. Our results show how entomological surveillance conducted around viraemic travellers can be effective for early detection of DENV in mosquitoes and thus might help to prevent possible autochthonous transmission

Effect of Salmosan, a β-galactomannan-rich product on intestinal barrier function

Podeu consultar el III Workshop anual INSA-UB complet a: http://hdl.handle.net/2445/118993Sessió 2. Comunicació oral núm.

Telomerase reverse transcriptase promoter mutations in bladder cancer: High frequency across stages, detection in urine, and lack of association with outcome

Background Hotspot mutations in the promoter of the gene coding for telomerase reverse transcriptase (TERT) have been described and proposed to activate gene expression. Objectives To investigate TERT mutation frequency, spectrum, association with expression and clinical outcome, and potential for detection of recurrences in urine in patients with urothelial bladder cancer (UBC). D