Toxicity assessment of pharmaceutical compounds on mixed culture from activated sludge using respirometric technique: The role of microbial community structure.

Micropollutants of emerging concern such as pharmaceuticals can significantly affect the performance of secondary biological processes in wastewater treatment plants. The present study is aimed to evaluate the toxicity and inhibition of three pharmaceutical compounds (caffeine, sulfamethoxazole and carbamazepine) on two cultures of microbial consortia enriched from wastewater aerobic activated sludge. One of them was acclimated to pharmaceuticals and the other was non-acclimated as control bioassay. The toxic and inhibitory effects on these cultures were assessed by respirometric tests through the oxygen uptake rate as an indicator of their capacity to degrade a readily available carbon source. Higher values of toxicity and inhibition of pharmaceutical compounds were observed for the control culture as compared to the acclimated one. Sulfamethoxazole and carbamazepine exhibited higher toxicity and inhibition effects than caffeine in both acclimated and control cultures. The microbial diversity of the two cultures was also studied. The composition of microbial community of acclimated and control cultures, was determined by targeting the 16S ribosomal RNA gene. It was observed that Proteobacteria was the most abundant phylum, with Gammaproteobacteria dominating both cultures. Control culture was dominated by Gammaproteobacteria and mostly by the genera Pseudomonas and Sodalis, which belong to common families present in wastewater. Results suggested that the acclimated culture to the three pharmaceuticals was mostly comprised of the extremely multiresistant genera Escherichia-Shigella (38%) of Gammaproteobacteria, resulting to higher resistance as compared to the control culture (Escherichia-Shigella, 7%). Finally, the microbial structure of the microorganisms present in a real bioreactor, which was initially seeded with the acclimated culture and fed in a continuous mode with the selected pharmaceuticals, was also analyzed. The continuous loading of pharmaceuticals in the bioreactor affected its microbial diversity, leading to the dominance of Betaproteobacteria and to the resistant genus Rhizobium of Alphaproteobacteria.

Biological removal of pharmaceutical compounds using white-rot fungi with concomitant FAME production of the residual biomass.

The efficiency of two white-rot fungi (WRF), Trametes versicolor and Ganoderma lucidum, to eliminate thirteen pharmaceutical pollutants with concomitant biodiesel production from the accumulating lipid content after treatment, was examined. The removal efficiency was studied using both individual and combined strains. The results of individual and combined strains showed a total removal (100%) of diclofenac (DCF), gemfibrozil (GFZ), ibuprofen (IBP), progesterone (PGT) and ranitidine (RNT). Lower removals were achieved for 4-acetamidoantipyrin (AAA), clofibric acid (ACF), atenolol (ATN), caffeine (CFN), carbamazepine (CZP), hydrochlorothiazide (HCT), sulfamethoxazole (SMX) and sulpiride (SPD), although the combination of both strains enhanced the system's efficiency, with removals ranging from 15 to 41%. This increase of the removal efficiency when combining both strains was attributed to the interactions developed between them (i.e., competition). Results from enzymatic and cytochrome P450 examination suggested that both extracellular (laccase, MnP, LiP) and intracellular oxidation mechanisms participate in the biological removal of pharmaceuticals. On the other hand, the "green" potential of the fungal sludge generated during the biological removal process was assessed for biodiesel production by means of one-step direct (in-situ) transformation. This process consists of the simultaneous extraction and conversion of lipids contained in the sludge by catalytic esterification/transesterification using a robust acid heterogeneous Zr-SBA-15 catalyst. This catalytic system provided conversions close to 80% of the saponifiable fraction (including free fatty acids and glycerides) in the presence of high amount of impurities. The overall weight FAME yield, based on the initial dried mass, was close to 30% for both strains.

Experimental and modeling study on removal of pharmaceutically active compounds in rotating biological contactors.

The aim of this work was to study the biological removal of pharmaceutical compounds in rotating biological contactors (RBCs) under continuous operation. A two-stage RBC was used, providing a total surface area of 1.41 m(2). Four pharmaceuticals of different therapeutic classes; caffeine, sulfamethoxazole, ranitidine and carbamazepine, were studied. Six experimental scenarios were applied to the RBC-system by varying substrates' loadings (12-54 gCOD/d), volumetric flow rate (2-5L/d), and pharmaceuticals' concentration (20-50 µg/L). The different conditions resulted to different solid retention times (SRT: 7-21 d) in each scenario. The increase of SRT due to variations of the operating conditions seemed to have a positive effect on pharmaceuticals' removal. Likewise, a negative correlation was observed between substrates' loading and pharmaceuticals' removal. An increase of initial pharmaceuticals' concentration resulted to decrease of SRT and pharmaceuticals' removal, suggesting a toxic effect to the biofilm. The maximum removals achieved were greater than 85% for all pharmaceuticals. Finally, a mathematical model which includes biofilm growth, substrates' utilization and pharmaceuticals' elimination was developed. The model predicts the contribution of sorption and biodegradation on pharmaceuticals' elimination taking into account the diffusion of pharmaceuticals inside biofilm.

Macrophages as potential targets for zoledronic acid outside the skeleton-evidence from in vitro and in vivo models.

PURPOSE: Multiple cell types of the tumour microenvironment, including macrophages, contribute to the response to cancer therapy. The anti-resorptive agent zoledronic acid (ZOL) has anti-tumour effects in vitro and in vivo, but it is not known to what extent macrophages are affected by this agent. We have therefore investigated the effects of ZOL on macrophages using a combination of in vitro and in vivo models. METHODS: J774 macrophages were treated with ZOL in vitro, alone and in combination with doxorubicin (DOX), and the levels of apoptosis and necrosis determined. Uptake of zoledronic acid was assessed by detection of unprenylated Rap1a in J774 macrophages in vitro, in peritoneal macrophages and in macrophage populations isolated from subcutaneously implanted breast cancer xenografts following ZOL treatment in vivo. RESULTS: Exposure of J774 macrophages to 5 µM ZOL for 24 h caused a significant increase in the levels of uRap1A, and higher doses/longer exposure induced apoptotic cell death. DOX (10 nM/24 h) and ZOL (10 µM/4 h) given in sequence induced significantly increased levels of apoptotic cell death compared to single agents. Peritoneal macrophages and macrophage populations isolated from breast tumour xenografts had detectable levels of uRap1A 24 h following a single, clinically achievable dose of 100 µg/kg ZOL in vivo. CONCLUSION: We demonstrate that macrophages are sensitive to sequential administration of DOX and ZOL, and that both peritoneal and breast tumour associated macrophages rapidly take up ZOL in vivo. Our data support that macrophages may contribute to the anti-tumour effect of ZOL.

Modeling of oleaginous fungal biofilm developed on semi-solid media.

An oleaginous fungus, Mortierella isabellina, able to transform efficiently sugar to storage lipid, was used as a model microorganism which develops a biofilm structure during the semi-solid fermentation process for the production of biodiesel from sweet sorghum. A mathematical model was developed to describe the fungal oil production in M. isabellina biofilm. The model describes diffusion and consumption of sugars and nitrogen of sweet sorghum and single cell oil production in a biofilm, which grows according to the kinetics of double-substrate limitation (sugars and nitrogen) with sugar inhibition. Experimental data from a previous experimental study were used to determine the kinetic parameters of the model. Maximum biofilm thickness and the percentage of lipid inside the biofilm were estimated using the model at 1892 µm and 15%, respectively. The proposed mathematical model could prove a useful tool for designing semi-solid fermentation processes.

Hydrogenotrophic denitrification of potable water: a review.

Several approaches of hydrogenotrophic denitrification of potable water as well as technical data and mathematical models that were developed for the process are reviewed. Most of the applications that were tested for hydrogenotrophic process achieved great efficiency, high denitrification rates, and operational simplicity. Moreover, this paper reviews the variety of reactor configurations that have been used for hydrogen gas generation and efficient hydrogen delivery. Microbial communities and species that participate in the denitrification process are also reported. The variation of nitrate concentration, pH, temperature, alkalinity, carbon and microbial acclimation was found to affect the denitrification rates. The main results regarding research progress on hydrogenotrophic denitrification are evaluated. Finally, the commonly used models and simulation approaches are discussed.

Experimental and modelling study of drinking water hydrogenotrophic denitrification in packed-bed reactors.

The aim of this work was to study hydrogenotrophic denitrification in packed-bed reactors under draw-fill and continuous operation. Three bench-scale packed-bed reactors with gravel in different sizes (mean diameter 1.75, 2.41 and 4.03 mm) as support media were used, in order to study the effect of particle size on reactors performance. The maximum denitrification rate achieved under draw-fill operation was 4.4 g NO(3)(-)-N/ld for the filter with gravel of 2.41 mm. This gravel size was chosen to perform experiments under continuous operation. Feed NO(3)(-)-N concentrations and hydraulic loadings (HL) ranged between 20-200mg/l and 5.7-22.8m(3)/m(2)d, respectively. A comparison between the two operating modes showed that, for low HL the draw-fill operation achieved higher denitrification rates, while for high HL and intermediate feed concentrations (40-60 mg NO(3)(-)-N/l) the continuous operation achieved higher denitrification rates (4.67-5.65 g/ld). Finally, experiments with three filters in series (with gravels of 4.03, 2.41 and 1.75 mm mean diameter) were also performed under continuous operation. The maximum denitrification rate achieved was 6.2 g NO(3)(-)-N/ld for feed concentration of 340 mg/l and HL of 11.5m(3)/m(2)d. A model, which describes denitrification in packed-bed reactors, was also developed. The model predicts the concentration profiles of NO(3)(-)-N along filter height, in draw-fill as well as in continuous operation, satisfactorily.

Kinetics of pure cultures of hydrogen-oxidizing denitrifying bacteria and modeling of the interactions among them in mixed cultures.

In this study we report the isolation of four denitrifying bacteria from a batch reactor, where the progress of hydrogenotrophic denitrification was examined. Only three of the strains had the ability to use hydrogen as electron donor. In the present work, kinetic batch experiments were carried out in order to study the dynamic characteristics of pure and defined mixed cultures of hydrogen-oxidizing denitrifying bacteria, under anoxic conditions, in a defined synthetic medium, in the presence of nitrates. Kinetic models were developed and the kinetic parameters were determined from the batch experiments for each bacterium separately. The behavior of mixed cultures and the interactions between the bacteria were described using kinetic models based on the kinetic models developed for each bacterium separately and their predictions were compared with the results from mixed culture experiments. The mathematical models that were developed and validated in the present work are capable of describing the behavior of the bacteria in pure and mixed cultures, and in particular, the kinetics of nitrate and nitrite reduction and cell growth.

Allotypes and gynecological cancer.

PURPOSE: To investigate if gynecologic cancer patients have different immune responses. METHODS: Immunoglobulin heavy chains (Glm, G3m), Kappa light chains (Km) and allotype and phenotype frequencies were examined in 58 patients with gynecologic cancer (ovarian, cervical, endometrial, vulval, vaginal and uterine sarcoma) and a control group of 26 women. RESULTS: No significant differences were found between the different allotype of phenotype frequencies between the two groups. CONCLUSIONS: Our results indicate that Gm and Km allotypes do not represent susceptibility factors for gynecologic cancer in Caucasians.