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Objectives: Bacterial viability and enrichment of resistance resulting from three different amikacin administration schedules with respect to haemodialysis (HD) were assessed against three OXA-48-producing Klebsiella pneumoniae isolated during an outbreak in a Spanish hospital., Methods: A previously described two-compartment dynamic system was used. Three possible amikacin administration schedules were simulated: (i) haemodialysis immediately after amikacin infusion (pre-HD); (ii) infusion immediately after haemodialysis (post-HD); and (iii) infusion at 50% interdialytic period. Amikacin standard dose (SD) and double dose (DD) were simulated for each schedule. Values of C max /MIC, C max /MPC (mutant prevention concentration), AUC 0-48h /MIC, AUC 0-48h /MPC and %T MSW (percentage of time that the concentration was inside the mutant selection window) were determined with experimental data and were correlated with the area under the bacterial killing curve of the total population and the resistant subpopulation., Results: Both with SD and DD, the pre-HD schedule resulted in increases at 48h in bacterial counts of the total population at the expense of enrichment of pre-existing resistant subpopulations from 12h onwards for all strains. The estimated %T MSW that prevented enrichment of resistant mutants was <61.5%. The AUC 0-48h /MPC (with values of ≈40 being associated with countering of increases in resistant subpopulations) was better than the %T MSW as a predictive parameter., Conclusion: This study showed that the longest times concentrations were above the MPC (i.e. highest AUC 0-48h /MPC, lowest %T MSW ), the lowest enrichment of resistant subpopulations. This implies use of the highest possible amikacin dose (limited by toxicity) and post-HD as the best administration schedule., (Copyright © 2019. Published by Elsevier Ltd.)

Acidiphilium spp. are conspicuous dwellers of acidic, metal-rich environments. Indeed, they are among the most metal-resistant organisms; yet little is known about the mechanisms behind the metal tolerance in this genus. Acidiphilium sp. PM is an environmental isolate from Rio Tinto, an acidic, metal-laden river located in southwestern Spain. The characterization of its metal resistance revealed a remarkable ability to tolerate high Ni concentrations. Here we report the screening of a genomic library of Acidiphilium sp. PM to identify genes involved in Ni resistance. This approach revealed seven different genes conferring Ni resistance to E. coli, two of which form an operon encoding the ATP-dependent protease HslVU (ClpQY). This protease was found to enhance resistance to both Ni and Co in E. coli, a function not previously reported. Other Ni-resistance determinants include genes involved in lipopolysaccharide biosynthesis and the synthesis of branched amino acids. The diversity of molecular functions of the genes recovered in the screening suggests that Ni resistance in Acidiphilium sp. PM probably relies on different molecular mechanisms.

Background: Whey protein isolate edible films with oregano or clove essential oils (EOs) incorporated as natural antimicrobials have been developed, with the aim of enhancing the microbial quality of poultry. The effectiveness of the films was determined against both the whole and selected microbiota developed during different periods of cold storage on the surface of skinless chicken breast. Tests were conducted by using both turbidimetric and agar disc diffusion methods., Results: The antimicrobial edible films developed showed high effectiveness against the main spoilers developed on the surface of skinless chicken breasts cold-stored for 8 days. The films based on oregano EO showed greater effectiveness than those based on clove EO. Still, clove EO could be part of an effective antimicrobial edible film. Enterobacteriaceae was the most susceptible to the effect of the films when lower concentrations of EO were incorporated. The largest inhibition surfaces obtained were provoked by films with the highest concentration of oregano EO incorporated against lactic acid bacteria., Conclusion: The antimicrobial edible films developed in this study inhibited the growth of the microbial populations that developed through storage of the chicken breast and caused its spoilage. The results of this research have direct application in the food industry to enhance the control of the development of spoilers such as Pseudomonas spp. or lactic acid bacteria., (© 2013 Society of Chemical Industry.)

Background: The use of aromatic plants and their extracts with antimicrobial properties may be compromised in the case of cheese, as some type of fungal starter is needed during its production. Penicillium verrucosum is considered a common cheese spoiler. The aim of this study was to evaluate the innovative use of certain aromatic plants as natural cheese covers in order to prevent mycotoxicogenic fungal growth (P. verrucosum). A collection of 12 essential oils (EOs) was obtained from various aromatic plants by solvent-free microwave extraction technology, and volatile characterisation of the EOs was carried out by gas chromatography/mass spectrometry., Results: The most effective EOs against P. verrucosum were obtained from Anethum graveolens, Hyssopus officinalis and Chamaemelum nobile, yielding 50% inhibition of fungal growth at concentration values lower than 0.02 µL mL⁻¹. All EOs showed high volatile heterogeneity, with α-phellandrene, pinocamphone, isopinocamphone, α-pinene, camphene, 1,8-cineole, carvacrol and trans-anethole being found to be statistically significant in the antifungal model., Conclusion: The use of these aromatic plants as natural covers on cheese can satisfactorily inhibit the growth of some mycotoxicogenic fungal spoilers. Among the volatile compounds present, α- and β-phellandrene were confirmed as the most relevant in the inhibition., (© 2012 Society of Chemical Industry.)

Confocal laser scanning microscopy (CLSM) helps to observe the biofilms formed in the endotracheal tube (ETT) of ventilated subjects and to determine its structure and bacterial viability using specific dyes. We compared the effect of three different treatments (placebo, linezolid, and vancomycin) on the bacterial biofilm viability captured by CLSM. Eight pigs with pneumonia induced by methicillin-resistant Staphylococcus aureus (MRSA) were ventilated up to 96 h and treated with linezolid, vancomycin, or placebo (controls). ETT images were microscopically examined after staining with the live/dead(®) BacLight(™) Kit (Invitrogen, Barcelona, Spain) with a confocal laser scanning microscope. We analyzed 127 images obtained by CLSM. The median ratio of live/dead bacteria was 0.51, 0.74, and 1 for the linezolid, vancomycin, and control groups, respectively (P = 0.002 for the three groups); this ratio was significantly lower for the linezolid group, compared with the control group (P = 0.001). Images showed bacterial biofilm attached and non-attached to the ETT surface but growing within secretions accumulated inside ETT. Systemic treatment with linezolid is associated with a higher proportion of dead bacteria in the ETT biofilm of animals with MRSA pneumonia. Biofilm clusters not necessarily attach to the ETT surface., (© 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

Background: The fresh-cut industry commonly uses sodium hypochlorite (NaClO) for disinfection. However, there are certain problems related to its use, and acidified sodium chlorite (ASC) could be an alternative sanitiser to replace it. There is limited research evaluating the effect of ASC on the overall quality of fresh-cut produce, especially sensory quality. In this study the decontamination efficacy and quality attribute effects of ASC on fresh-cut tatsoi after application and during storage were investigated., Results: Tatsoi baby leaves were minimally processed at 8 °C and stored under passive modified atmosphere packaging for up to 11 days at 5 and 10 °C. Low to moderate doses of ASC (100-500 mg L⁻¹) showed an initial antimicrobial efficacy on natural microflora and Escherichia coli as effective as that of NaClO. Regarding contact time, ASC was effective in reducting the E. coli population during the first 30 s of washing, and an increase in contact time did not improve the antimicrobial effect. Sensory quality attributes were well kept for up to 11 days at 5 °C but for only 5 days at the abusive temperature of 10 °C., Conclusion: ASC provides an alternative sanitising technique to NaClO for maintaining the quality and safety of fresh-cut tatsoi baby leaves for up to 11 days at 5 °C., (Copyright © 2011 Society of Chemical Industry.)

The aim of this work was to study the effect of olive powder combined with high hydrostatic pressure (HHP) on the inactivation of Bacillus cereus spores, to use it as an additional control hurdle in beverages pasteurised by this technology. With this purpose, reference medium prepared at different concentrations of olive powder was inoculated with B. cereus spores and subjected to different pressure treatments. The outgrowth capacity of the treated spores was then determined at 20°C and 32°C. The addition of olive powder was found to slightly reduce the effectiveness of HHP, although in post-treatment storage there was an increased bacteriostatic effect in the samples with 2.5% of olive powder at both temperatures in the samples pressurised at 400 and 500 MPa, and only at 20°C in the samples pressurised at 200 MPa. The addition of olive powder therefore had an additive effect with storage temperature and HHP processing and could act as an additional control hurdle during the shelf-life of products pasteurised by HHP technologies or in the case of cold-chain breakage.