Assessment of an aerobic granular sludge system in the presence of pharmaceutically active compounds by quantitative image analysis and chemometric techniques.

In this study, a sequencing batch reactor (SBR) with aerobic granular sludge (AGS) was operated with synthetic wastewater containing environmental relevant concentrations of 17ß-estradiol (E2), 17α-ethinylestradiol (EE2) and sulfamethoxazole (SMX). Despite the presence of the studied PhAC, the granular fraction clearly predominated (TSSgran/TSS ranging from 0.82 to 0.98) throughout the monitoring period, presenting aggregates with high organic fraction (VSS/TSS above 0.83) and good settling characteristics (SVI5 ranging from 15 to 39 mL/gTSS). A principal component analysis (PCA) with quantitative image analysis (QIA) based data allowed to distinguish the different operational periods, namely with mature granules (CONT), and the E2, EE2, and SMX feeding periods. It further revealed a positive relationship between the biomass density, sludge settling ability, overall and granular biomass contents, granulation properties, granular biomass fraction and large granules fraction and size. Moreover, a discriminant analysis (DA) allowed to successfully discriminate not only the different operational periods, mainly by using the floccular apparent density, granular stratification and contents data, but also the PhAC presence in samples. The filamentous bacteria contents, sludge settling properties, settling properties stability and granular stratification, structure and contents parameters were found to be crucial for that purpose.

Use of chemometric analyses to assess biological wastewater treatment plants by protozoa and metazoa monitoring.

Protozoa and metazoa biota communities in biological wastewater treatment plants (WWTP) are known to be dependent of both the plant type (oxidation ditch, trickling filter, conventional activated sludge, among others) and the working operational conditions (incoming effluent characteristics, toxics presence, organic load, aeration, hydraulic and sludge retention times, nitrification occurrence, etc.). Thus, for analogous WWTP operating in equivalent operating conditions, similar protozoa and metazoa communities can be found. Indeed, the protozoa and metazoa biota monitoring can be considered a quite useful tool for assessing the functioning of biological WWTP. Furthermore, the use of chemometric techniques in WWTP monitoring is becoming widespread to enlighten interrelationships within the plant, especially when a large collection of data can be obtained. In the current study, the protozoa and metazoa communities of three different types of WWTP, comprising one oxidation ditch, four trickling filters, and three conventional activated sludge plants, were monitored. For that purpose, metazoa, as well as the main protozoa groups (flagellates, free-swimming, crawling and sessile ciliates, and testate amoeba) were determined in terms of contents and relative abundance. The collected data was further processed by chemometric techniques, such as cross-correlation, principal components, multivariate ANOVA, and decision trees analyses, allowing to successfully identify, and characterize, the different studied WWTP, and thus, being able to help monitoring and diagnosing operational problems.

Quantitative image analysis of polyhydroxyalkanoates inclusions from microbial mixed cultures under different SBR operation strategies.

Polyhydroxyalkanoates (PHAs) produced from mixed microbial cultures (MMC), regarded as potential substitutes of petrochemical plastics, can be found as intracellular granules in various microorganisms under limited nutrient conditions and excess of carbon source. PHA is traditionally quantified by laborious and time-consuming chromatography analysis, and a simpler and faster method to assess PHA contents from MMC, such as quantitative image analysis (QIA), is of great interest. The main purpose of the present work was to upgrade a previously developed QIA methodology (as reported by Mesquita et al. (Anal Chim Acta 770:36-44, 2013a, Anal Chim Acta 865:8-15, 2015)) for MMC intracellular PHA contents quantification, increase the studied intracellular PHA concentration range, and extend to different sequencing batch reactor (SBR) operation strategies. Therefore, the operation of a new aerobic dynamic feeding (ADF) SBR allowed further extending the studied operating conditions, dataset, and range of the MMC intracellular PHA contents from the previously reported anaerobic/aerobic cycle SBR. Nile Blue A (NBA) staining was employed for epifluorescence microscope visualization and image acquisition, further fed to a custom developed QIA. Data from each of the feast and famine cycles of both SBR were individually processed using chemometrics analysis, obtaining the correspondent partial least squares (PLS) models. The PHA concentrations determined from PLS models were further plotted against the results obtained in the standard chromatographic method. For both SBR, the predicted ability was higher at the end of the feast stage than for the famine stage. Indeed, an independent feast and famine QIA data treatment was found to be fundamental to obtain the best prediction abilities. Furthermore, a promising overall correlation (R 2 of 0.83) could be found combining the overall QIA data regarding the PHA prediction up to a concentration of 1785.1 mg L-1 (37.3 wt%). Thus, the results confirm that the presented QIA methodology can be seen as promising for estimating higher intracellular PHA concentrations for a larger reactors operation systems and further extending the prediction range of previous studies.

Microbial-based evaluation of foaming events in full-scale wastewater treatment plants by microscopy survey and quantitative image analysis.

Activated sludge systems are prone to be affected by foaming occurrences causing the sludge to rise in the reactor and affecting the wastewater treatment plant (WWTP) performance. Nonetheless, there is currently a knowledge gap hindering the development of foaming events prediction tools that may be fulfilled by the quantitative monitoring of AS systems biota and sludge characteristics. As such, the present study focuses on the assessment of foaming events in full-scale WWTPs, by quantitative protozoa, metazoa, filamentous bacteria, and sludge characteristics analysis, further used to enlighten the inner relationships between these parameters. In the current study, a conventional activated sludge system (CAS) and an oxidation ditch (OD) were surveyed throughout a period of 2 and 3 months, respectively, regarding their biota and sludge characteristics. The biota community was monitored by microscopic observation, and a new filamentous bacteria index was developed to quantify their occurrence. Sludge characteristics (aggregated and filamentous biomass contents and aggregate size) were determined by quantitative image analysis (QIA). The obtained data was then processed by principal components analysis (PCA), cross-correlation analysis, and decision trees to assess the foaming occurrences, and enlighten the inner relationships. It was found that such events were best assessed by the combined use of the relative abundance of testate amoeba and nocardioform filamentous index, presenting a 92.9 % success rate for overall foaming events, and 87.5 and 100 %, respectively, for persistent and mild events.

Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining.

Polyhydroxyalkanoates (PHA) can be produced and intracellularly accumulated as inclusions by mixed microbial cultures (MMC) for bioplastic production and in enhanced biological phosphorus removal (EBPR) systems. Classical methods for PHA quantification use a digestion step prior to chromatography analysis, rendering them labor intensive and time-consuming. The present work investigates the use of two quantitative image analysis (QIA) procedures specifically developed for PHA inclusions identification and quantification. MMC obtained from an EBPR system were visualized by bright-field and fluorescence microscopy for PHA inclusions detection, upon Sudan Black B (SBB) and Nile Blue A (NBA) staining, respectively. The captured color images were processed by QIA techniques and the image analysis data were further treated using multivariate statistical analysis. Partial least squares (PLS) regression coefficients of 0.90 and 0.86 were obtained between QIA parameters and PHA concentrations using SBB and NBA, respectively. It was found that both staining procedures might be seen as alternative methodologies to classical PHA determination.

Monitoring intracellular polyphosphate accumulation in enhanced biological phosphorus removal systems by quantitative image analysis.

A rapid methodology for intracellular storage polyphosphate (poly-P) identification and monitoring in enhanced biological phosphorus removal (EBPR) systems is proposed based on quantitative image analysis (QIA). In EBPR systems, 4',6-diamidino-2-phenylindole (DAPI) is usually combined with fluorescence in situ hybridization to evaluate the microbial community. The proposed monitoring technique is based on a QIA procedure specifically developed for determining poly-P inclusions within a biomass suspension using solely DAPI by epifluorescence microscopy. Due to contradictory literature regarding DAPI concentrations used for poly-P detection, the present work assessed the optimal DAPI concentration for samples acquired at the end of the EBPR aerobic stage when the accumulation occurred. Digital images were then acquired and processed by means of image processing and analysis. A correlation was found between average poly-P intensity values and the analytical determination. The proposed methodology can be seen as a promising alternative procedure for quantifying intracellular poly-P accumulation in a faster and less labour-intensive way.

Prediction of intracellular storage polymers using quantitative image analysis in enhanced biological phosphorus removal systems.

The present study focuses on predicting the concentration of intracellular storage polymers in enhanced biological phosphorus removal (EBPR) systems. For that purpose, quantitative image analysis techniques were developed for determining the intracellular concentrations of PHA (PHB and PHV) with Nile blue and glycogen with aniline blue staining. Partial least squares (PLS) were used to predict the standard analytical values of these polymers by the proposed methodology. Identification of the aerobic and anaerobic stages proved to be crucial for improving the assessment of PHA, PHB and PHV intracellular concentrations. Current Nile blue based methodology can be seen as a feasible starting point for further enhancement. Glycogen detection based on the developed aniline blue staining methodology combined with the image analysis data proved to be a promising technique, toward the elimination of the need for analytical off-line measurements.

O impacto emocional da cirurgia bariátrica em pacientes com obesidade mórbida / The emotional impact of bariatric surgery in patients with morbid obesity

A cirurgia bariátrica é um método que tem sido utilizado para o emagrecimento e resgate da saúde. Em muitos casos, porém, o emagrecimento súbito incorre em quadros psiquiátricos, tais como: sintomas depressivos, ansiedade, uso de substâncias, alterações comportamentais e ideação suicida, dentre outras situações, associadas às mudanças emocionais que o paciente vivencia com o novo estado físico e psíquico. O atual estudo teve como objetivo analisar as expectativas, fantasias, resultados, dificuldades e frustrações enfrentadas após o procedimento cirúrgico. Realizou-se um relato de caso das informações colhidas com seis pacientes em acompanhamento num hospital público de Santa Catarina e que desenvolveram sintomas de sofrimento psíquico após a cirurgia bariátrica. Cada paciente participou de duas sessões de entrevistas semi-estruturadas. A pesquisa foi submetida e aprovada pela comissão de ética da Universidade de Joinville. Os resultados obtidos demonstram que as mudanças psicológicas decorrentes da cirurgia foram marcantes. Em determinados casos, as entrevistadas expressaram expectativas além do emagrecimento, como a resolução dos conflitos interpessoais e conjugais, assim como mudanças de traços definidos de suas personalidades. Constatou-se, também, o uso de substâncias (álcool e tabaco) associado a comportamentos de risco (envolvimento extraconjugal e direção perigosa). O problema da obesidade ficou evidenciado como parte de uma complexa situação que envolve o estado físico e emocional das pessoas, e sua resolução, por vezes, expõe as dificuldades e as limitações psíquicas dos pacientes.

Bariatric surgery has been used for health recovery and weight loss. In many cases, however, abrupt weight loss produces psychiatric symptoms, such as depression, anxiety, use of drugs, behavioral changes and suicidal ideation, among other situations, all related to the emotional changes faced by the patient in his new physical and chemical state. This study aimed at analyzing hopes, fantasies, results, difficulties, and frustrations present after a surgical procedure. This case report resulted from the data collected from six patients being followed at a public hospital in Santa Catarina, Brazil, who developed symptoms of psychic suffering after the bariatric surgery. Each patient participated in two sessions of semi-structured interviews. This research was approved by the ethics committee of Universidade de Joinville. Results show that psychological changes resulting from the bariatric surgery were remarkable. In some cases, patients expressed expectations beyond weight loss, such as resolution of marital and interpersonal conflicts, as well as changes in stable personality traits. Use of substances (alcohol and tobacco) associated with risk behavior (extra-marital relationships and dangerous driving) was also detected. The problem of obesity was evidenced as part of a complex situation that involves both physical and mental status, and its solution sometimes exposes the patient's difficulties and psychological limitations.