Polyethersulfone (PES) Filters Improve the Recovery of Legionella spp. and Enhance Selectivity against Interfering Microorganisms in Water Samples.

In the analysis of water samples, the type of filtration membrane material can influence the recovery of Legionella species, although this issue has been poorly investigated. Filtration membranes (0.45 µm) from different materials and manufacturers (numbered as 1, 2, 3, 4, and 5) were compared: mixed cellulose esters (MCEs), nitrocellulose (NC), and polyethersulfone (PES). After membrane filtration of samples, filters were placed directly onto GVPC agar and incubated at 36 ± 2 °C. The highest mean counts of colony-forming units and colony sizes for Legionella pneumophila and Legionella anisa were obtained with PES filters (p < 0.001). All membranes placed on GVPC agar totally inhibited Escherichia coli and Enterococcus faecalis ATCC 19443 and ATCC 29212, whereas only the PES filter from manufacturer 3 (3-PES) totally inhibited Pseudomonas aeruginosa. PES membrane performance also differed according to the manufacturer, with 3-PES providing the best productivity and selectivity. In real water samples, 3-PES also produced a higher Legionella recovery and better inhibition of interfering microorganisms. These results support the use of PES membranes in methods where the filter is placed directly on the culture media and not only in procedures where membrane filtration is followed by a washing step (according to ISO 11731:2017).

Use of Blood Powder (Ground and Irradiated) for the Manufacture of Chocolate Agar.

Chocolate agar (CA) is an enriched medium for the isolation and identification of fastidious bacteria. Defibrinated blood is used to manufacture CA, but this expensive product is not always affordable for companies in developing countries. Blood powder (BP) is potentially a cheaper alternative, although its pre-treatment using autoclaving can impair the quality of the media. Therefore, optimization of BP as a substitute for defibrinated blood for CA manufacture deserves further research. CA was manufactured with irradiated BP (dehydrated bovine blood powder) and its physical and microbiological characteristics were compared with those of conventional CA and CA prepared with autoclaved BP. Each medium was seeded with 20-200 CFU of target bacteria using the spiral pouring method. Finally, another medium was prepared using BP pre-treated by grinding and gamma irradiation and its performance assessed. Compared to conventional CA, the medium containing ground and irradiated BP provided a similar CFU count for both fastidious (Neisseria, Haemophilus, Campylobacter, and Streptococcus) and non-fastidious (Moraxella, Staphylococcus, Enterococcus, Klebsiella, and Pseudomonas) species, unlike the medium prepared with BP subjected only to irradiation, which provided a lower growth of fastidious species. Morphology and characteristics of all bacterial colonies were very similar in conventional CA and the new medium, the number of Pseudomonas CFU being higher in the latter. The medium prepared with ground plus irradiated vs. irradiated BP more closely resembled conventional CA, having a browner background. The new CA medium prepared with ground and gamma irradiation-sterilized BP has comparable productivity properties to conventional CA. Therefore, it could be a more practical and economical methodology to facilitate large-scale CA manufacture.

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

Glycine-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.

Radical scavenging of white tea and its flavonoid constituents by electron paramagnetic resonance (EPR) spectroscopy.

White tea (WT) presents high levels of catechins, which are known to reduce oxidative stress. WT is the least processed tea, unfermented and prepared only from very young tea leaves. The subject of this paper is the use of the spin trap method and electron paramagnetic resonance (EPR) spectroscopy as the analytical tool to measure, for the first time, the radical scavenging activity of WT and its major catechin components, epicatechin (EC), epicatechin-3-gallate (ECG), epigallocatechin (EGC), and epigallocatechin-3-gallate (EGCG), against the methoxy radical, using ferulic acid as antioxidant pattern. The antioxidant activity has been measured by the decrease of the intensity of the spectral bands of the adduct DMPO-OCH3 in the EPR with the amount of antioxidant in the reactive mixture. Tea leaves and buds were extracted with waterless methanol. It has been proved that tea compounds with more antiradical activity against methoxy radical are those with the gallate group, EGCG and ECG.

Lactobacillus plantarum CECT 7527, 7528 and 7529: probiotic candidates to reduce cholesterol levels.

BACKGROUND: Hypercholesterolaemia plays a key role in the development and progression of coronary artery disease. There are numerous drugs available to treat this condition but they are often expensive and can have unwanted side effects. Therefore, a screening of lactic acid bacteria to select candidate probiotic strains to reduce cholesterol levels was carried out. Three Lactobacillus plantarum strains (CECT 7527, 7528 and 7529) were selected as potential probiotics to reduce cholesterol levels after conducting several in vitro tests for demonstrating the functionalities of the strains according to international guidelines. RESULTS: The three strains showed a high ability to survive under gastrointestinal tract conditions and to adhere to intestinal cells. Regarding lipid metabolism, the strains showed a great production of bile salt hydrolase, especially when combined. Moreover, the strains assimilated cholesterol directly from the medium. Part of the cholesterol present in the medium was removed via binding onto the bacterial cellular surface. Finally, the three strains, especially CECT 7529, produce large quantities of propionic and butyric acids. CONCLUSION: Combined, these characteristics suggest that these strains could be excellent candidates for reducing high blood cholesterol levels.

Antioxidant properties of aqueous and ethanolic extracts of tara (Caesalpinia spinosa) pods in vitro and in model food emulsions.

BACKGROUND: The successful replacement of some synthetic food antioxidants by safe natural antioxidants has fostered intensive search for new vegetable sources of antioxidants. In our study the phenol and flavonoid content of extracts of tara pods was determined. The antioxidant activity was also studied by three different analytical assays: the measurement of scavenging capacity against a radical ABTS⁺ , the oxygen radical absorbance capacity (ORAC) and the ferric reducing antioxidant power (FRAP). RESULTS: All analyzed samples showed a good antioxidant capacity, but the use of a solution of ethanol 75% in a 1 h ultrasonic process allowed achieving the greatest quantity of phenolics (0.464 mg gallic acid equivalent (GAE) g⁻¹ dry weight (DW) ) and the highest antioxidant activity measured by the ABTS⁺ and ORAC methods (10.17 and 4.29 mmol L⁻¹ Trolox equivalents (TE) g⁻¹ DW, respectively). The best method for efficient extraction of flavonoids (3.08 mg catechin equivalent (CE) g⁻¹ DW) was a 24 h maceration in cold water. Two extracts obtained with ethanol 75% and water were added to a model food system (oil-in-water emulsion) and the oxidative stability was studied during storage at 38 °C. Oxidation was monitored by determination of the peroxide value. The addition of 48 µg mL⁻¹ ethanol extract to the emulsion delayed oxidation to the same extent as 17.8 µg mL⁻¹ of Trolox, while water extract was only effective in the early stages of the oxidation process. CONCLUSION: The results of this study indicate that ethanolic tara extracts may be suitable for use in food, cosmetic and nutraceutical applications.

Extraction of Antioxidants from Borage (Borago officinalis L.) Leaves-Optimization by Response Surface Method and Application in Oil-in-Water Emulsions.

Borage (Borago officinalis L.) is a typical Spanish plant. During processing, 60% are leaves. The aim of this work is to model and optimize the extraction of polyphenol from borage leaves using the response surface method (RSM) and to use this extract for application in emulsions. The responses were: total polyphenol content (TPC), antioxidant capacity by ORAC, and rosmarinic acid by HPLC. The ranges of the variables temperature, ethanol content and time were 50-90 °C, 0%-30%-60% ethanol (v/v), and 10-15 min. For ethanolic extraction, optimal conditions were at 75.9 °C, 52% ethanol and 14.8 min, yielding activity of 27.05 mg GAE/g DW TPC; 115.96 mg TE/g DW in ORAC and 11.02 mg/L rosmarinic acid. For water extraction, optimal activity was achieved with extraction at 98.3 °C and 22 min, with responses of 22.3 mg GAE/g DW TPC; 81.6 mg TE/g DW in ORAC and 3.9 mg/L rosmarinic acid. The significant variables were ethanol concentration and temperature. For emulsions, the peroxide value was inhibited by 60% for 3% extract concentration; and 80% with 3% extract concentration and 0.2% of BSA. The p-anisidine value between the control and the emulsion with 3% extract was reduced to 73.6% and with BSA 86.3%, and others concentrations had similar behavior.

Modelling Extraction of White Tea Polyphenols: The Influence of Temperature and Ethanol Concentration.

The optimization of the extraction of natural antioxidants from white tea has fostered intensive research. This study has investigated the effects of ethanol-water mixtures, temperature and time on the extraction of polyphenols and antioxidant components from white tea. The response surface methodology was applied to identify the best extraction conditions. The best conditions to maximize the extraction of total polyphenols were: ethanol, 50%, for 47.5 min. Although the yield of polyphenols was optimal at 65 °C, the maximum antioxidant capacity was achieved with an extraction temperature of 90 °C. This study has identified the optimal conditions for the extraction of tea liquor with the best antioxidant properties. Epigallocatechin gallate, epicatechin gallate, epigallocatechin and epicatechin were extracted from white tea at concentrations up to 29.6 ± 10.6, 5.40 ± 2.09, 5.04 ± 0.20 and 2.48 ± 1.10 mg/100 g.