EMA-amplicon-based sequencing informs risk assessment analysis of water treatment systems.

Illumina amplicon-based sequencing was coupled with ethidium monoazide bromide (EMA) pre-treatment to monitor the total viable bacterial community and subsequently identify and prioritise the target organisms for the health risk assessment of the untreated rainwater and rainwater treated using large-volume batch solar reactor prototypes installed in an informal settlement and rural farming community. Taxonomic assignments indicated that Legionella and Pseudomonas were the most frequently detected genera containing opportunistic bacterial pathogens in the untreated and treated rainwater at both sites. Additionally, Mycobacterium, Clostridium sensu stricto and Escherichia/Shigella displayed high (≥80%) detection frequencies in the untreated and/or treated rainwater samples at one or both sites. Numerous exposure scenarios (e.g. drinking, cleaning) were subsequently investigated and the health risk of using untreated and solar reactor treated rainwater in developing countries was quantified based on the presence of L. pneumophila, P. aeruginosa and E. coli. The solar reactor prototypes were able to reduce the health risk associated with E. coli and P. aeruginosa to below the 1 × 10-4 annual benchmark limit for all the non-potable uses of rainwater within the target communities (exception of showering for E. coli). However, the risk associated with intentional drinking of untreated or treated rainwater exceeded the benchmark limit (E. coli and P. aeruginosa). Additionally, while the solar reactor treatment reduced the risk associated with garden hosing and showering based on the presence of L. pneumophila, the risk estimates for both activities still exceeded the annual benchmark limit. The large-volume batch solar reactor prototypes were thus able to reduce the risk posed by the target bacteria for non-potable activities rainwater is commonly used for in water scarce regions of sub-Saharan Africa. This study highlights the need to assess water treatment systems in field trials using QMRA.

Antimicrobial activity of Citrus spp. and Anethum graveolens components against Candida metapsilosis in ranch sauce.

In this paper, a study was carried out to test the inhibitory effect of a natural food compound (NFC), based on flavonoids (naringenin, hesperetin, tangeritin, luteolin, apigenin and kaempferol) from citrus and dill, in ranch sauce. A strain of C. metapsilosis, isolated from a spoiled sample of ranch sauce, was used as target pathogen microorganism. The inhibitory effect of NFC was compared with a common mixture of chemical preservatives used in this type of sauces: potassium sorbate and sodium benzoate (S/B). An in vitro test was performed by the microtiter plate assay at 10, 25 and 37 °C for 24 h in modified Tryptic Soy Broth. An additive antimicrobial effect had been observed in the combination of acetic acid and NFC. The results of the microtiter assay were validated in a challenge test in ranch sauce at 5, 25 and 37 °C for 10 weeks. NFC showed partial fungicidal effect against C. metapsilosis, reducing two logarithmic units at 5 °C for 10 weeks. At 5 °C, the traditional doses of S/B used in ranch sauce decreased viable cells to non-detectable counts from the second week of the experiment. At 25 and 37 °C, the use of S/B mixture or the use of NFC showed the same fungicidal effect. The incorporation of NFC, alone or in combination with acetic acid, opens the possibility of formulating clean label sauces with good protection against the development of the acid resistant yeast C. metapsilosis.

Validation of large-volume batch solar reactors for the treatment of rainwater in field trials in sub-Saharan Africa.

The efficiency of two large-volume batch solar reactors [Prototype I (140 L) and II (88 L)] in treating rainwater on-site in a local informal settlement and farming community was assessed. Untreated [Tank 1 and Tank 2-(First-flush)] and treated (Prototype I and II) tank water samples were routinely collected from each site and all the measured physico-chemical parameters (e.g. pH and turbidity, amongst others), anions (e.g. sulphate and chloride, amongst others) and cations (e.g. iron and lead, amongst others) were within national and international drinking water guidelines limits. Culture-based analysis indicated that Escherichia coli, total and faecal coliforms, enterococci and heterotrophic bacteria counts exceeded drinking water guideline limits in 61%, 100%, 45%, 24% and 100% of the untreated tank water samples collected from both sites. However, an 8 hour solar exposure treatment for both solar reactors was sufficient to reduce these indicator organisms to within national and international drinking water standards, with the exception of the heterotrophic bacteria which exceeded the drinking water standard limit in 43% of the samples treated with the Prototype I reactor (1 log reduction). Molecular viability analysis subsequently indicated that mean overall reductions of 75% and 74% were obtained for the analysed indicator organisms (E. coli and enterococci spp.) and opportunistic pathogens (Klebsiella spp., Legionella spp., Pseudomonas spp., Salmonella spp. and Cryptosporidium spp. oocysts) in the Prototype I and II solar reactors, respectively. The large-volume batch solar reactor prototypes could thus effectively provide four (88 L Prototype II) to seven (144 L Prototype I) people on a daily basis with the basic water requirement for human activities (20 L). Additionally, a generic Water Safety Plan was developed to aid practitioners in identifying risks and implement remedial actions in this type of installation in order to ensure the safety of the treated water.

[Clinical Psychologist in Primary Care: The work carried out in Asturias]. / Psicología Clínica en Atención Primaria: la experiencia en Asturias.

AIM: To describe the characteristics of the population attended in the clinical psychology programme in Primary Care, as well as to perform a preliminary analysis of the results obtained. METHOD: Prospective, descriptive study of the results obtained after brief psychological intervention. The following evaluation tools were used: PHQ-9, GAD-7, WHOQOL-BREF, and an ad hoc designed satisfaction questionnaire. RESULTS: A total of 1084 patients were referred to the program, and were mostly women. The most frequent diagnoses were anxiety, adaptive disorders, Zcodes (factors that influence the state of health and contact with health services) and depressive disorders. The mean waiting list time was 7.25days, and the mean number of consultations was3. The analyses performed after the treatment showed an improvement in all the variables evaluated, as well as high satisfaction and usefulness of the treatment. CONCLUSIONS: The integration of the clinical psychologist in Primary Care leads to an improvement in the quality of care of patients with emotional disorders, facilitating an interdisciplinary bio-psycho-social approach, and offering internationally recommended evidence-based treatments and interventions.

Microbiological evaluation of combined advanced chemical-biological oxidation technologies for the treatment of cork boiling wastewater.

This paper contains a multidisciplinary approach that will contribute to design and properly evaluate a treatment line for complex biorecalcitrant wastewaters. To demonstrate this approach a specific industrial wastewater (cork boiling wastewater, CBW) was used. A treatment line based on a coagulation-flocculation step followed by an Advanced Oxidation Process (AOP) (solar photo-Fenton) and combined with an aerobic biological system was evaluated. Applied microbiological techniques: optical microscopy, plate count, DNA extraction and qPCR, indicated that some communities disappeared after the activated sludge adaptation period to the partially treated wastewater, while communities that did not disappear were damaged: 2-log reduction of total heterotrophic bacteria (THB) and a decrease in DNA concentration from 200 ng/µL to 65 ng/µL were observed. Therefore, chemical and microbiological results obtained along the set of experiments, suggested the inefficiency of the combined treatment option between solar photo-Fenton and advanced aerobic biological systems for CBW. This led to the necessity of applying solar photo-Fenton without combining with biotreatment and with the objective of improving the effluent quality enough for being reused in the own industry. Toxicity tests, based on different organisms (after coagulation-flocculation followed by solar photo-Fenton), showed increase on acute toxicity (from 46% to 71% by respirometric assays) and the development of chronic toxicity (from 21-29% to 83-90% also measured by respirometric assays), made evident the incompatibility of this type of wastewater with a biological treatment even after the application of an AOP.

A pilot study into the use of Continuous Venous Hyperfiltration to manage patients in a critical state with dysregulated inflammation. / Estudio piloto sobre el uso de la Hiperfiltración Venosa Continua para el manejo de pacientes en estado crítico con inflamación desregulada.

INTRODUCTION: Haemofiltration paradigms used to manage critically ill patients with a dysregulated inflammatory response (DIR) assess kidney function to monitor its onset, adaptation, and completion. A Continuous Venous Hyperfiltration (CONVEHY) protocol is presented, in which a non-specific adsorption membrane (AN69-ST-Heparin Grafted) is used with citrate as an anticoagulant and substitution fluid. CONVEHY uses tools readily available to achieve kidney related and non-related objectives, and it is guided by the monitoring of pathophysiological responses. OBJECTIVES: To compare the response to an AN69-ST-HG membrane when heparin (He, n=5: Standard protocol) or citrate (Ci, n=6: CONVEHY protocol) was used to evaluate whether a larger study into the benefits of this protocol would be feasible. MATERIALS AND METHODS: In a retrospective pilot study, the benefits of the CONVEHY protocol to manage patients with a DIR in a surgical critical care unit (CCUs) were assessed by evaluating the SOFA (Sequential Organ Failure Assessment) (He 11 ± 2.35; Ci 11 ± 3.63: p=0.54) and APACHE II (He 28.60 ± 9.40; Ci 24 ± 8.46: p=0.93) scores. RESULTS: Nights in hospital (He 35.2 ± 16.3 nights; Ci 9 ± 2.53: p=0.004), hospital admission after discharge from the CCUs (He 40.25 ± 21.82; Ci 13.2 ± 4.09: p=0.063), patients hospitalised >20 days (He 80%; Ci 0%: p=0.048), days requiring mechanical ventilation (He 16 ± 5.66; Ci 4 ± 1.72: p=0.004), and the predicted (55.39 ± 26.13%) versus real mortality in both groups (9.1%: p=0.004). CONCLUSIONS: The CONVEHY protocol improves the clinical responses of patients with DIR, highlighting the potential value of performing larger and confirmatory studies.

Identification of transformation products of carbamazepine in lettuce crops irrigated with Ultraviolet-C treated water.

Transformation of organic microcontaminants (OMCs) during wastewater treatments results in the generation of transformation products (TPs), which can be more persistent than parent compounds. Due to reuse of reclaimed wastewater (RWW) for crop irrigation, OMCs and TPs are released in soils being capable to translocate to crops. Furthermore, OMCs are also susceptible to transformation once they reach the soil or crops. The recalcitrant antiepileptic carbamazepine (CBZ) and some of its frequently reported TPs have been found in agricultural systems. However, there is no knowledge about the fate in reuse practices of multiple CBZ TPs that can be formed during wastewater treatment processes. For the first time, this work presents a study of the behavior of CBZ TPs generated after a conventional Ultraviolet-C (UVC) treatment in an agricultural environment. The UVC-treated water was used for the irrigation of lettuces grown under controlled conditions. The latter was compared to the fate of TPs generated in the peat and plant by irrigation with non-treated water containing CBZ. A suspect screening strategy was developed to identify the TPs using liquid chromatography coupled to quadrupole-time-of-flight (LC-QTOF-MS). The results revealed the presence of 24 TPs, 22 in UVC-treated water, 11 in peat and 9 in lettuce leaves. 4 of the TPs identified in peat (iminostilbene, TP 271B, TP 285A-B); and 3 in leaves (10-11 dihydrocarbamazepine, TP 271A-B) were not previously reported in soils or edible parts of crops, respectively. Comparing the TPs found in peat and lettuces derived from both irrigation conditions, no significant differences regarding TPs formation or occurrence were observed. UVC treatment did not contribute to the formation of different TPs than those generated by transformation or metabolism of CBZ in peat or plant material. This research improves the current knowledge on the fate of CBZ TPs in agricultural systems because of reuse practices.

Construction of a synthetic protein using PCR with a high essential amino acid content for nutritional purposes.

Ovalbumin is considered a protein of high nutritional value because it contains essential amino acids and is highly digestible. Therefore, it has a high biological value. Currently, the high food demand requires worldwide attention because food production is insufficient. Therefore, other alternatives are necessary to satisfy food demands, such as protein engineering. In this work, a protein with a high essential amino acid content similar to ovalbumin was synthesized by protein engineering, expressed, and digested in vitro. The assembly and sequential overlap extension PCR strategy was used to synthesize a 345-bp gene that encodes a high essential amino acid content protein (HEAAP). The 345-bp product was cloned into the vector pBAD TOPO®, and expressed in Escherichia coli BL21. PCR reactions and sequencing demonstrated the presence, orientation, and correct sequence of the insert. HEAAP expression was induced by L-arabinose and then purified using Ni-NTA affinity chromatography. The expression in E. coli was low and barely detected by Western blot assay. The in vitro multienzyme digestibility of HEAAP was around 79%, which suggests that the protein is potentially nutritious. Virtual analysis classifies the protein as unstable and hydrophilic, with a half-life in E. coli of 10 h. The recombinant HEAAP was successfully synthesized, but it is necessary to improve the digestibility and to optimize expression including selecting other expression models.

Practical approach to the evaluation of industrial wastewater treatment by the application of advanced microbiological techniques.

In cork industry, the operation of boiling raw cork generates large volumes of wastewater named Cork Boiling Wastewater (CBW). The main characteristics are the low biodegradability and medium to low acute toxicity, resulting in the necessity of designing advanced biological treatments by possible conventional activated sludge adaptation. In order to evaluate the variation of bacterial population along that process, a study based on optical microscopy, plate count, DNA extraction, qPCR and massive sequencing techniques was performed. Results showed a diminution of the total and volatile solids (TSS and VSS), jointly with a decrease in DNA concentration, general bacteria (16 S) and ammonia-oxidizing bacteria (AOB). After a few hours of testing, diverse microbiological species died while others showed a possible adaptation of the biological system, accompained by a dissolved organic carbon (DOC) reduction. In addition, toxicity tests based on activated sludge showed the development of chronic toxicity through the contact time. Combination of classical and advanced microbiological techniques, such as quantitative real time Polymerase Chain Reaction (qPCR) and metagenomics, was essential to predict the variation of species during the experiment and to conclude if effective biological adaptation could be finally attained for the target complex wastewater.

Validation and application of a multiresidue method based on liquid chromatography-tandem mass spectrometry for evaluating the plant uptake of 74 microcontaminants in crops irrigated with treated municipal wastewater.

Reuse of treated wastewater for agricultural purposes can mitigate water stress in some regions where the lack of water is an extended problem. However, the environmental long-term consequences of this practice are still unknown. It is demonstrated that using reclaimed water for irrigation lead to accumulation and translocation of some microcontaminants (MCs) in soil and crops. However, so far, only a small group of contaminants has been investigated. This study aims to develop and validate a simple and efficient multiresidue method based on QuEChERs (Quick, Easy, Cheap, Effective and Rugged) extraction coupled to liquid chromatography tandem mass spectrometry (LC-MS/MS). The novelty of the study relays in the large number of MCs analyzed (74), some of them not previously investigated, in three commodities (lettuce, radish and strawberry). Optimized conditions yielded good results for the three commodities under study. Up to 84% of the compounds were recovered within a 70-120% range, with good repeatability (relative standard deviations below 20% in most cases). Method detection (MDLs) and quantification limits (MQLs) ranged from 0.01 to 2 ng/g. The proposed method was successfully applied to assess the potential uptake of MCs by lettuce and radish crops irrigated with wastewater under controlled conditions for 3 and 1.5 months, respectively. 12 compounds were detected in the crops with concentrations ranging from 0.03 to 57.6 ng/g. N-Formyl-4-aminoantipyrine (4FAA) was the most concentrated compound. The application of this method demonstrated for the first time the accumulation of 5 contaminants of emerging concern (CECs) not previously reported: 4FAA, N-Acetyl-4-aminoantipyrine (4AAA), hydrochlorothiazide, mepivacaine and venlafaxine.

Disinfection of water inoculated with Enterococcus faecalis using solar/Fe(III)EDDS-H2O2 or S2O82- process.

In this study, the activation of H2O2 and persulfate ions induced by solar photolysis of Fe(III)EDDS complex were investigated in water disinfection, applying solar AOPs processes. The use of Fe(III)EDDS complex maintains iron in soluble form until slightly basic pH and so the photolysis is efficient in a large range of pH compatible with natural waters. Moreover, for the first time, the impact of photogenerated hydroxyl and sulfate radicals on the inactivation of Enterococcus faecalis in water was studied. E. faecalis was proposed as alternative model microorganism given its higher resistance than the commonly used E. coli. The reactivity of hydroxyl radicals seems to be more efficient for the inactivation of such strain than the reactivity of sulfate radicals. Moreover, experimental results show that the concentration of Fe(III)EDDS complex is a key parameter for the inactivation of microrganisms. For the direct application in natural waters, the efficiency of the process in the presence of ubiquitous inorganic compounds, such as carbonate (HCO3-/CO32-) and chloride ions (Cl-), was also investigated. Carbonates showed a strong reduction on the E. faecalis inactivation in all cases; meanwhile chloride ions enhanced the inactivation in the presence of persulfate as also shown by using a complementary kinetic modeling approach. A dual role of Fe(III)EDDS complex was established and discussed; essential for the generation of radical species but a trap for the reactivity of these same radicals.

Integration of Membrane Distillation with solar photo-Fenton for purification of water contaminated with Bacillus sp. and Clostridium sp. spores.

Although Membrane Distillation (MD) has been extensively studied for desalination, it has other applications like removing all kinds of solutes from water and concentrating non-volatile substances. MD offers the possibility of producing a clean stream while concentrating valuable compounds from waste streams towards their recovery, or emerging contaminants and pathogens present in wastewater in order to facilitate their chemical elimination. This paper analyses the elimination of bacterial spores from contaminated water with MD and the role of MD in the subsequent treatment of the concentrate with photo-Fenton process. The experiments were performed at Plataforma Solar de Almería (PSA) using a plate and frame bench module with a Permeate Gap Membrane Distillation (PGMD) configuration. Tests were done for two different kinds of spores in two different water matrixes: distilled water with 3.5wt% of sea salts contaminated with spores of Bacillus subtilis (B. subtilis) and wastewater after a secondary treatment and still contaminated with Clostridium sp. spores. An analysis of the permeate was performed in all cases to determine its purity, as well as the concentrated stream and its further treatment in order to assess the benefits of using MD. Results showed a permeate free of spores in all the cases, demonstrating the viability of MD to treat biological contaminated wastewater for further use in agriculture. Moreover, the results obtained after treating the concentrate with photo-Fenton showed a shorter treatment time for the reduction of the spore concentration in the water than that when only photo-Fenton was used.

[New devices of fecal diversion and collection: adverse effects described in the literature]. / Nuevos dispositivos de derivación y colección fecal: efectos adversos descritos en la bibliografía.

UNLABELLED: Among products for the management and control of faeces in patients with fecal incontinence, there are new devices that allow the diversion and collection of fecal matter. This study has aimed to know the complications related to these new devices described in the literature. METHOD: A search was made in the main bibliographic databases, obtaining publications on the new devices. The relevant documents were selected, these being those that described complications. After these were described. RESULTS: A total of 13 relevant documents were recovered. There were 45 adverse events 36 patients. Those having the greatest incidence were rectal bleeding (10 patients) and abdominal distension (6 patients). CONCLUSION: In order to provide the best patient care, it is essential to know the possible complications in order to be able to prevent them. The cares given these patients need to be reevaluated.