Response of Anaerobic Protozoa to Oxygen Tension in Anaerobic System.

The effect of oxygen on anaerobic protozoa was studied in anaerobic batch reactors inoculated with sludge and protozoa cultures. Among the protozoa genera, Metopus, Brachonella, Plagiopyla, Trepomonas, and Vanella were more sensitive to oxygen compared to other genera. Protozoa genera Menoidium, Rhynchomonas, Cyclidium, Spathidium, and Amoeba were found to survive under aerobic conditions, and the growth rate was slightly higher or similar to anaerobic condition. O2 tension resulted in the loss of free and endosymbiotic methanogens in anaerobic system, while methanogens were observed inside the protozoan cysts. Survival of anaerobic protozoa declined considerably when the O2 tension exceeded 1% atm. sat. and showed chemosensory behavior in response to O2 exposure. Superoxide dismutase activity was detected in survived protozoa cells under O2 tension. Facultative anaerobic protozoa with SOD activity can provide a mechanism to overcome possible occurrence of oxygen toxicity in the treatment of wastewater in anaerobic reactor.

Large-scale cultivation of Leishmania infantum promastigotes in stirred bioreactor.

BACKGROUND & OBJECTIVES: Bioreactors are practical tools that are used for economical, time-conserving and large-scale production of biomass from cell cultivation. They provide optimal environmental conditions such as pH and temperature required for obtaining maximum amounts of biomass. However, there is no evidence in the literature on the large-scale cultivation of Leishmania infantum parasites in the bioreactor. Therefore, the present study was undertaken to develop a new approach for obtaining L. infantum biomass by using pH and temperature controllable stirred bioreactor and to compare parasitic growth kinetics with classical method within erlenmeyers. METHODS: In order to obtain parasite biomass, a newly developed pH and temperature controlled stirred bioreactor was used and its efficacy was compared with a graduated classical scale-up method. Growth kinetics of parasites within erlenmeyers and bioreactors were determined by evaluating promastigote numbers using haemocytometer. The graduated scale enlargement of culture was followed by T25 flask, T75 flask, and 1 L erlenmeyer, respectively. RESULTS: Obtained results showed a 10-fold increase in the number of promastigotes within the conventional culture performed in 700 ml medium, while parasite numbers increased approximately 15 times due to initial inoculation amounts in the bioreactor culture performed in the 3.5 l medium. Thus, there was 7.5 times more biomass collection in bioreactor compared to classical method. INTERPRETATION & CONCLUSION: It is postulated that constant culture pH and temperature in the bioreactor extends cultivation time. This could lead to significant increase in parasite numbers. Hence, pH and temperature controllable bioreactors provided acquisition of sufficient amounts of biomass in contrast to classical methods. Therefore, this type of bioreactors may substitute classical culture methods in the production of antigenic molecules for vaccine development.

A bioreactor system for the manufacture of a genetically modified Plasmodium falciparum blood stage malaria cell bank for use in a clinical trial.

BACKGROUND: Although the use of induced blood stage malaria infection has proven to be a valuable tool for testing the efficacy of vaccines and drugs against Plasmodium falciparum, a limiting factor has been the availability of Good Manufacturing Practice (GMP)-compliant defined P. falciparum strains for in vivo use. The aim of this study was to develop a cost-effective method for the large-scale production of P. falciparum cell banks suitable for use in clinical trials. METHODS: Genetically-attenuated parasites (GAP) were produced by targeted deletion of the gene encoding the knob associated histidine rich protein (kahrp) from P. falciparum strain 3D7. A GAP master cell bank (MCB) was manufactured by culturing parasites in an FDA approved single use, closed system sterile plastic bioreactor. All components used to manufacture the MCB were screened to comply with standards appropriate for in vivo use. The cryopreserved MCB was subjected to extensive testing to ensure GMP compliance for a phase 1 investigational product. RESULTS: Two hundred vials of the GAP MCB were successfully manufactured. At harvest, the GAP MCB had a parasitaemia of 6.3%, with 96% of parasites at ring stage. Testing confirmed that all release criteria were met (sterility, absence of viral contaminants and endotoxins, parasite viability following cryopreservation, identity and anti-malarial drug sensitivity of parasites). CONCLUSION: Large-scale in vitro culture of P. falciparum parasites using a wave bioreactor can be achieved under GMP-compliant conditions. This provides a cost-effective methodology for the production of malaria parasites suitable for administration in clinical trials.

Eukaryotic Community Shift in Response to Organic Loading Rate of an Aerobic Trickling Filter (Down-Flow Hanging Sponge Reactor) Treating Domestic Sewage.

In this study, changes in eukaryotic community structure and water quality were investigated in an aerobic trickling filter (down-flow hanging sponge, DHS) treating domestic sewage under different organic loading rates (OLRs). The OLR clearly influenced both sponge pore water quality and relative flagellates and ciliates (free-swimming, carnivorous, crawling, and stalked protozoa) abundances in the retained sludge. Immediately after the OLR was increased from 1.05 to 1.97 kg chemical oxygen demand (COD) m-3 day-1, COD and NH4+-N treatment efficiencies both deteriorated, and relative flagellates and ciliates abundances then increased from 2-8 % to 51-65 % total cells in the middle-bottom part of the DHS reactor. In a continuous operation at a stable OLR (2.01 kg COD m-3 day-1), effluent water quality improved, and relative flagellates and ciliates abundances decreased to 15-46 % total cells in the middle-bottom part of the DHS reactor. This result may indicate that flagellates and ciliates preferentially graze on dispersed bacteria, thus, stabilizing effluent water quality. Additionally, to investigate eukaryotic community structure, clone libraries based on the 18S ribosomal ribonucleic acid (rRNA) gene of the retained sludge were constructed. The predominant group was Nucletmycea phylotypes, representing approximately 29-56 % total clones. Furthermore, a large proportion of the clones had <97 % sequence identity in the NCBI database. This result indicates that phylogenetically unknown eukaryotes were present in the DHS reactor. These results provide insights into eukaryotic community shift in the DHS reactor treating domestic sewage.

Putrescine: Essential factor for in vitro proliferation of Babesia bovis.

This study reports the effect of putrescine addition, either alone or in combination with insulin, transferrin and selenite (ITS), to serum-free Advanced DMEM/F12 (A-DMEM/F12) medium, on the in vitro culture of Babesia bovis and using a perfusion bioreactor to improve efficiency of the process. A B. bovis strain previously adapted to proliferate in serum-free medium (Bbovis-SF) was evaluated using eight increasing concentrations of putrescine. The percentage of parasitized erythrocytes (PPE) obtained from cultures supplemented with 0.101 mg/L was 6.23% compared with 2.3% for control cultures with M199 with Earle's salts (M199) and 40% serum. The combination of putrescine (0.101 mg/L) and a mixture of ITS (2000, 1100, and 1.34 mg/L, respectively) (Pu-ITS), in A-DMEM/F12 culture medium without serum yielded a maximum PPE of 17.26% compared to 2.58% in the control medium. This new formulation of culture medium, together with the use of a hollow-fiber perfusion bioreactor system (HFPBS), caused a substantial increase in the proliferation of B. bovis, yielding a maximum cumulative PPE of 118.8% after five days, compared to 58.6% in cultures treated with control medium M199 and 40% serum. We concluded that the addition of the ITS mixture and putrescine to the culture medium stimulated the proliferation of B. bovis in vitro. This new medium formulation, used in a HFPBS culture system, can be an effective, automated-prone system that can induce massive proliferation of B. bovis for use as a source of parasite antigens and immunogens.

Prokaryote species richness is positively correlated with eukaryote abundance in wastewater treatment biofilms.

Biological treatment represents a key step in nutrient removal from wastewater. Until now these process has mainly been considered prokaryotic, with the interactions between prokaryotes and eukaryotes not being properly explored. We therefore investigated the co-occurrence of eukaryotes and prokaryotes in biological nitrogen removal biofilms. We found that biofilms in the nitrifying reactor contained the highest diversity and abundance of both prokaryotes and eukaryotes, with nearly three times higher prokaryote species richness than for the denitrifying reactor. The positive associations between eukaryote abundance and prokaryote diversity could potentially be explained by mutualism - and/or predator/prey interactions. Further mechanistic insight, however, is needed to determine the main diversifying mechanisms. In summary, eukaryote and prokaryote interactions seem to play a fundamental yet underexplored role in biological wastewater treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: Eukaryote and prokaryote interactions may play an important role in wastewater treatment. This study found that prokaryote species richness was nearly three times higher in the aerobe nitrification than in an anaerobe denitrification reactor, coinciding with the highest level of eukaryotes. This knowledge can be important in process control, and potentially in the development of novel approaches based on nitrate accumulating denitrifying eukaryotes.

The components of shear stress affecting insect cells used with the baculovirus expression vector system.

Insect-based expression platforms such as the baculovirus expression vector system (BEVS) are widely used for the laboratory- and industrial-scale production of recombinant proteins. Thereby, major drawbacks to gain high-quality proteins are the lytic infection cycle and the shear sensitivity of infected insect cells due to turbulence and aeration. Smaller bubbles were formerly assumed to be more harmful than larger ones, but we found that cell damage is also dependent on the concentration of protective agents such as Pluronic®. At the appropriate concentration, Pluronic forms a layer around air bubbles and hinders the attachment of cells, thus limiting the damage. In this context, we used microaeration to vary bubble sizes and confirmed that size is not the most important factor, but the total gas surface area in the reactor is. If the surface area exceeds a certain threshold, the concentration of Pluronic is no longer sufficient for cell protection. To investigate the significance of shear forces, a second study was carried out in which infected insect cells were cultivated in a hollow fiber module to protect them from shear forces. Both model studies revealed important aspects of the design and scale-up of BEVS processes for the production of recombinant proteins.

Assessing the fate of Ascaris suum ova during mesophilic anaerobic digestion.

There is limited knowledge about the survival of geohelminths, which are soil-transmitted human pathogens, in mesophilic anaerobic digestion processes. This study examined the fate of embryonated and unembryonated Ascaris suum ova in six laboratory-scale mesophilic (35 °C) anaerobic digesters processing swine manure to identify their survival strategies and investigate potential mechanisms to enhance their destruction. There was no significant difference in inactivation of Ascaris suum ova in digesters operated at different solids residence times (SRT) or feeding frequencies. Ova exposed to an anaerobic environment became dormant, or remained unembryonated throughout their residence in the reactors. Approximately 65% of ova were able to retain their viability for up to 16 days, after which the rate of inactivation increased until nearly all ova were nonviable by day 24. In contrast, ova exposed to aerobic conditions did not become dormant and progressed through several developmental stages until day 16, after which nearly all ova were observed to be nonviable. In addition, only 35% of fully developed ova exposed to the anaerobic environment retained their viability by day 16 compared to 65% for dormant ova. Results suggest that some ova are physically destroyed during digestion and ova can be inactivated faster if their development cycle is aerobically triggered before entering the anaerobic digestion process. Results also suggest that transfer of resource recovery technologies such as mesophilic anaerobic digestion to developing world settings must account for local climatic and health conditions so mutually beneficial outcomes can be attained.

Microbiological and performance evaluation of sequencing batch reactor for textile wastewater treatment.

This study focused on laboratory-scaled and real-scaled treatment plant performances and microbiological investigations for the optimum treatment of textile industry wastewater performed with sequencing batch reactor (SBR). As a result of experimental studies of laboratory-scaled SBR treatment unit, optimum treatment efficiency was taken from 0.5 h filling to 1.5 h. reaction to 1.5 h. settlement to 0.5 h. discharge-idle periods. Average chemical oxygen demand (COD) removal efficiency of SBR of laboratory-scaled textile industry was 75%, whereas average turbidity and color removal (coloration number [RES, m(-1)] 586 nm) efficiencies were 90% and 75%, respectively. Optimum reaction and settlement periods were used in a real-scaled plant, and plant efficiency was examined for parameters such as COD, phenol, pH, mixed liquor suspended solids (MLSS) and sludge volume index (SVI). In this study, optimum reaction and settlement periods for treatment of textile industry wastewater were determined within a SBR in a laboratory-scaled plant. These reaction and settlement periods were verified with the measurement of COD, color, and turbidity parameters. Floc structure and protozoa-metazoa species of activated sludge in a SBR were also determined. Optimum reaction and settlement times were used in a real-scaled plant, and plant efficiency was examined for COD, Phenol, pH, MLSS, and SVI parameters. The corresponding values were found as appropriate, acceptable, and meaningful because of variance value of statistical analysis. Protozoa and metazoan in the activated sludge in the laboratory-scaled plant were investigated. Peranema sp., Epistylis sp., Didinium sp., Chilodonella sp., Opercularia sp., Vorticella sp. as protozoa species and Habrotrocha sp., Philodina sp. as metazoa species were determined.

In Vitro Culture of Cryptosporidium parvum Using Hollow Fiber Bioreactor: Applications for Simultaneous Pharmacokinetic and Pharmacodynamic Evaluation of Test Compounds.

Hollow fiber technology is a powerful tool for the culture of difficult-to-grow cells. Cryptosporidium parvum has a multistage sexual and asexual life cycle that has proved difficult to culture by conventional in vitro culture methods. Here, we describe a method utilizing a hollow fiber bioreactor for the continuous in vitro growth of C. parvum that produces sexual and asexual stages. The method enables the evaluation of potential therapeutic compounds under conditions that mirror the dynamic conditions found in the gut facilitating preliminary pharmacokinetic and pharmacodynamic data to be obtained.

Specialized proteomic responses and an ancient photoprotection mechanism sustain marine green algal growth during phosphate limitation.

Marine algae perform approximately half of global carbon fixation, but their growth is often limited by the availability of phosphate or other nutrients1,2. As oceans warm, the area of phosphate-limited surface waters is predicted to increase, resulting in ocean desertification3,4. Understanding the responses of key eukaryotic phytoplankton to nutrient limitation is therefore critical5,6. We used advanced photo-bioreactors to investigate how the widespread marine green alga Micromonas commoda grows under transitions from replete nutrients to chronic phosphate limitation and subsequent relief, analysing photosystem changes and broad cellular responses using proteomics, transcriptomics and biophysical measurements. We find that physiological and protein expression responses previously attributed to stress are critical to supporting stable exponential growth when phosphate is limiting. Unexpectedly, the abundance of most proteins involved in light harvesting does not change, but an ancient light-harvesting-related protein, LHCSR, is induced and dissipates damaging excess absorbed light as heat throughout phosphate limitation. Concurrently, a suite of uncharacterized proteins with narrow phylogenetic distributions increase multifold. Notably, of the proteins that exhibit significant changes, 70% are not differentially expressed at the mRNA transcript level, highlighting the importance of post-transcriptional processes in microbial eukaryotes. Nevertheless, transcript-protein pairs with concordant changes were identified that will enable more robust interpretation of eukaryotic phytoplankton responses in the field from metatranscriptomic studies. Our results show that P-limited Micromonas responds quickly to a fresh pulse of phosphate by rapidly increasing replication, and that the protein network associated with this ability is composed of both conserved and phylogenetically recent proteome systems that promote dynamic phosphate homeostasis. That an ancient mechanism for mitigating light stress is central to sustaining growth during extended phosphate limitation highlights the possibility of interactive effects arising from combined stressors under ocean change, which could reduce the efficacy of algal strategies for optimizing marine photosynthesis.

Nematodes in wastewater biofilms--appearance and density of species in three biofilter reactors.

Population dynamics of nematode species in biofilms of three different biofilter reactors, differing in size (pilot/laboratory scale), operation mode and biofilm carrier, were studied over a period of 1 year. In the biofilm suspension of the pilot system mean nematode density was 118individuals/ml and average biomass 15microg wet weight/ml. Higher mean abundance was found in the two laboratory systems with 2380 and 4411individuals/ml. Mean biomass in the laboratory systems ranged from 209 to 330microg wet weight/ml. There were marked temporal differences in appearance and density of nematode species in all three biofilters. Number of species observed was 3 in the laboratory systems and 5 in the pilot system. The fastest growing species (Paroigolaimella bernensis and Diplogasteritus nudicapitatus) were observed in the pilot reactor in contrast to the more slowly growing species (Diploscapter coronatus and Acrostichus sp.), which dominated in the laboratory reactors. Sexual reproduction was found for all species but of Diploscapter coronatus. When comparing life history traits of the different species with the environmental conditions in the reactors, it seems that the unstable conditions in the pilot reactor favor the fast growing species whereas the stable environment in the laboratory systems allows the growth of species with longer generation times.

Full-scale class A biosolids production by two-stage continuous-batch thermophilic anaerobic digestion at the hyperion treatment plant, Los Angeles, California.

The City of Los Angeles Hyperion Treatment Plant (HTP) (California) converted its anaerobic digesters to thermophilic operation to produce Class A biosolids. Phase IV tests demonstrated compliance of a two-stage, continuous-batch process with Alternative 1 of U.S. Environmental Protection Agency 40 CFR Part 503 (U.S. EPA, 1993), which defines the time-temperature requirement for batch treatment (T > or = 56.3 degrees C at 16-h holding). Fecal coliforms, Salmonella sp., viable helminth ova, and enteric viruses were not detected in biosolids in the postdigestion train, including the truck-loading facility and the farm for land application as the last points of plant control where compliance is to be demonstrated. The same results were achieved during Phase V tests, after lowering the second-stage holding temperature to 52.6 degrees C to reduce the elevated methyl mercaptan production that was observed during Phase IV. Hence, the Phase V process complied with Alternative 3 of 40 CFR Part 503. Currently, HTP operates its digesters under the same conditions as tested in Phase V. In 2003, monthly monitoring of the biosolids at the truck-loading facility and the farm for land application demonstrated consistent compliance with Alternative 3.

Composting-vermicomposting of leaf litter ensuing from the trees of mango (Mangifera indica).

Litter of the mango (Mangifera indica) tree leaves was composted and then converted into vermicast by the action of the earthworm Eudrilus eugeniae Kinberg. After over nine months of continuous operation the vermireactors with 62.5 animals l(-1) generated approximately 13.6g vermicast per litre of reactor volume (l) per day (d) whereas the reactors with 75 animals l(-1) produced approximately 14.9 g vermicast l(-1) d(-1). This difference in performance of the reactors operating in duplicate at the two different earthworm densities was statistically significant (> or = 90% confidence level) for most of the nine-month span. The animals grew well in all reactors, increasing their zoomass by approximately 103% and producing approximately 157 offspring. Not a single of the 1100 animals died during the first four months. In the subsequent five months a total of 122 worms died, representing a loss of approximately 2% per month. We attribute this to the normal process of ageing. The ability of the earthworms to survive, grow and breed in the vermireactors fed with composted mango tree leaves, and a rising trend in vermicast output inspite of the death of a few worms after four months of reactor operation, indicate the sustainability of this type of vermireactors. The studies also indicate that even better vermireactor efficiency may be possible by modifying the reactor geometry. Studies on changes in C:N ratio during composting and vermicomposting revealed that whereas composting helped in lowering the ratio due to loss of carbon in bacterial metabolism, vermicomposting had no such effect on the ratio.

Vermistabilization of textile mill sludge spiked with poultry droppings by an epigeic earthworm Eisenia foetida.

Investigations were made to explore the potential of an epigeic earthworm Eisenia foetida to transform textile mill sludge spiked with poultry droppings in to value added product, i.e., vermicompost. The growth and reproduction of E. foetida was monitored in a range of different feed mixtures for 77 days in the laboratory under controlled experimental conditions. The maximum growth was recorded in 100% cow dung (CD). Replacement of poultry droppings by cow dung in feed mixtures and vice versa had little or no effect on worm growth rate and reproduction potential. Worms grew and reproduced favourably in 70% poultry droppings (PD)+30% solid textile mill sludge (STMS) and 60% PD+40% STMS feed mixtures. Greater percentage of STMS in the feed mixture significantly affected the biomass gain and cocoon production. Net weight gain by earthworms in 100% CD was 2.9-18.2 fold higher than different STMS containing feed mixtures. The mean number of cocoon production was between 23.4+/-4.65 (in 100% CD) and 3.6+/-1.04 (in 50% PD+50% STMS) cocoons earthworm(-1) for different feed mixtures tested. Vermicomposting resulted in significant reduction in C:N ratio and increase in nitrogen and phosphorus contents. Total potassium, total calcium and heavy metals (Fe, Zn, Pb and Cd) contents were lower in the final product than initial feed mixtures. Our trials demonstrated vermicomposting as an alternate technology for the recycling and environmentally safe disposal/management of textile mill sludge using an epigeic earthworm E. foetida if mixed with poultry droppings.

Enhanced degradation of waste grass clippings in one and two stage anaerobic systems.

The present work investigated the use of a simple rumen-fluid-inoculated anaerobic treatment system for the degradation of organic waste. Fresh rumen fluid collected from a fistulated sheep was used as the inoculum and fresh grass clippings were used as the waste material for treatment. Studies were carried out on both a one-stage system where the ligno-cellulosic fraction breaks down into a mixture of soluble products including volatile fatty acids and a two- stage system where these products are subsequently mineralised to biogas. In the one stage system about 70% of the organic waste was solubilized and in the two stage system about 60% waste material was solubilized in a week. About 50% of the degradation was achieved in a 4 day period, showing that a 4 day solids retention time would be a suitable operating regime. The maximum volatile fatty acid production rate was 327 mg COD l(-1) h(-1). A higher loading rate of 30 g l(-1) d(-1) was achieved in these systems compared to anaerobic digesters. Microbiological studies showed an increase in the number of fungal spores as well as a decrease in the number of protozoa in the treatment system. These numbers attained stable values over the duration of the experiments. The system developed is superior to conventional composting or anaerobic digestion and can be applied for the treatment of ligno-cellulosic agricultural residues.

Effect of high levels of the rotifer Lecane inermis on the ciliate community in laboratory-scale sequencing batch bioreactors (SBRs).

Due to its ability to feed on filamentous bacteria, the rotifer Lecane inermis has already been recognized as a potential control agent of activated sludge bulking, which is usually caused by the excessive growth of filamentous microorganisms. However, their effectiveness depends, in part, on their abundance. We studied the influence of high densities of L. inermis on the protozoan community in activated sludge from a wastewater treatment plant (WWTP) in 4 laboratory-scale sequencing batch bioreactors (SBRs). Two treatments and two controls were subjected to nutrient removal system in process similar to that used in a WWTP. The experiment lasted 9 days and was repeated in 24-h cycles, including phases of agitation with feeding, aeration and agitation and sedimentation with decantation at the end of the cycle. In total, 32 taxa were identified, among which 25 were ciliated protozoa, 4 were amoebae, 2 were flagellates, and one was a nematode. Rotifers were then introduced to 2 bioreactors at a final concentration of 500ind.mL(-1), and the taxonomic composition and abundance of the activated sludge microfauna were assessed 2, 5 and 8 days thereafter. The mean density of ciliates on the first day of experiment was 12,610ind.mL(-1) and diminished to 4868±432ind.mL-±432ind.mL(-1) in the control and 5496±638ind.mL(-1) in the rotifer-treated group on the last day. Thus, even extremely high densities of artificially introduced rotifers did not negatively affect the protozoan community. On the contrary, the protozoan community was more diverse in the treatment group than in the control.

NaCS-PDMDAAC immobilized cultivation of recombinant Dictyostelium discoideum for soluble human Fas ligand production.

Dictyostelium discoideum is a promising eukaryotic host for the expression of heterologous proteins requiring post-translational modifications. However, the dilute nature of D. discoideum cell culture limits applications for high value proteins production. D. discoideum cells, entrapped in sodium cellulose sulfate/poly-dimethyl-diallyl-ammonium chloride (NaCS-PDMDAAC) capsules were used for biosynthesis of the heterologous protein, soluble human Fas ligand (hFasL). Semi-continuous cultivations with capsules recycling were carried out in shake flasks. Also, a scaled-up cultivation of immobilized D. discoideum for hFasL production in a customized vitreous airlift bioreactor was conducted. The results show that NaCS-PDMDAAC capsules have desirable biophysical properties including biocompatibility with the D. discoideum cells and good mechanical stability throughout the duration of cultivation. A maximum cell density of 2.02 × 10(7) cells mL(-1) (equivalent to a maximum cell density of 2.22 × 10(8) cells mL(-1) in capsules) and a hFasL concentration of 130.40 µg L(-1) (equivalent to a hFasL concentration of 1434.40 µg L(-1) in capsules) were obtained in shake flask cultivation with capsules recycling. Also, a maximum cell density of 1.72 × 10(7) cells mL(-1) (equivalent to a maximum cell density of 1.89 × 10(8) cells mL(-1) in capsules) and a hFasL concentration of 106.10 µg L(-1) (equivalent to a hFasL concentration of 1167.10 µg L(-1) in capsules) were obtained after ∼170 h cultivation in the airlift bioreactor (with a working volume of 200 mL in a 315 mL bioreactor). As the article presents a premier work in the application of NaCS-PDMDAAC immobilized D. discoideum cells for the production of hFasL, more work is required to further optimize the system to generate higher cell densities and hFasL titers for large-scale applications.

Dynamics of nematodes in a high organic loading rotating biological contactors.

Nematode diversity and dynamics of a full-scale rotating biological contactor plant (RBC) has been studied. Analysis of biofilm composition showed a well-established zoning of microfauna among the three RBC sections analysed. Nematodes appeared to be the dominant group within the larger microfauna populations with average abundances between 200 and 300ind/mg or 8000 and 17000ind/cm(2). The most abundant nematode species were Diplogasteritus nudicapitatus and Paroigolaimella coprophages and, to a lesser extent, Paroigolaimella bernensis and Steinernema intermedia. The relationship between nematodes and filamentous bacteria (specifically the genus Beggiatoa) was the most significant biotic relationship found, and to a lesser extent, nematodes with ciliates. The relationship between the abundance of nematode species and the physical-chemical variables suggests that nematodes may be good indicators of low pollutant load levels in the entry of the RBC system. Finally, the results indicate that nematodes may have a relevant role for a good biofilm development.

Quantitative analysis of ammonia-oxidizing bacteria in a combined system of MBR and worm reactors treating synthetic wastewater.

The Static Sequencing Batch Worm Reactor (SSBWR) followed by the MBR (S-MBR) is one of the advanced excess sludge treatments. In this paper, the control MBR (C-MBR) and the SSBWR-MBR were operated in parallel to study the changes of NH3-N removal and ammonia oxidizing bacteria (AOB). The results showed that the capacity of NH3-N removal of the S-MBR was improved by the worm reactors along with the operation. The S-MBR was favorable because it selected for the higher activity of the ammonia oxidization and better cells appearance of the sludge. The five species (Nitrosomonas, Betaproteobacteria, Clostridium, Dechloromonas and Bacteria) were found to be significantly correlate with the ammonia oxidization functions and performance of NH3-N removal in the C-MBR and S-MBR. The Nitrosomonas, Betaproteobacteria and Dechloromonas remained and eventually enriched in the S-MBR played a primary role in the NH3-N removal of the S-MBR.