Virucidal activity of microalgae extracts harvested during phycoremediation of swine wastewater.

Phycoremediation of swine wastewater is a promising treatment since it efficiently removes nutrients and contaminants and, simultaneously, its biomass can be harvested and used to obtain a wide range of valuable compounds and metabolites. In this context, biomass microalgae were investigated for the phycoremediation of swine wastewater, and biomass extracts for its virucidal effect against enveloped and non-enveloped viruses. Microalgae were cultivated in a pilot scale bioreactor fed with swine wastewater as the growth substrate. Hexane, dichloromethane, and methanol were used to obtain the microalgae extracts. Extracts were tested for virucidal potential against HSV-1 and HAdV-5. Virucidal assays were conducted at temperatures that emulate environmental conditions (21 °C) and body temperature (37 °C). The maximum production of microalgae biomass reached a concentration of 318.5 ± 23.6 mgDW L-1. The results showed that phycoremediation removed 100% of ammonia-N and phosphate-P, with rates (k1) of 0.218 ± 0.013 and 0.501 ± 0.038 (day-1), respectively. All microalgae extract reduced 100% of the infectious capacity of HSV-1. The microalgae extracts with dichloromethane and methanol showed inhibition activities at the lowest concentration (3.125 µg mL-1). Virucidal assays against HAdV-5 using microalgae extract of hexane and methanol inhibited the infectious capacity of the virus by 70% at all concentrations tested at 37 °C. At a concentration of 12.5 µg mL-1, the dichloromethane microalgae extract reduced 50-80% of the infectious capacity of HAdV-5, also at 37 °C. Overall, the results suggest that the microalgae can be an attractive source of feedstock biomass for the exploration of alternative virucidal compounds.

Fagos de bactérias redutoras de sulfato em água de produção de um reservatório de petróleo offshore: método tentativo de concentração e purificação / Phages of Sulphate Reducing Bacteria in reactor water from offshore oil wells: tentative method for concentration and purification

RESUMO O controle do crescimento microbiano é um desafio crescente na produção de petróleo e gás, uma vez que a presença de determinadas bactérias traz impactos econômica e ambientalmente negativos. As bactérias redutoras de sulfato (BRS) são particularmente problemáticas, uma vez que são responsáveis pela corrosão biológica ligada à produção de sulfeto de hidrogênio, efeito conhecido como souring. A principal forma de controle das BRS atualmente é a injeção de biocidas, no entanto essa estratégia, além de requerer aplicação contínua, tem se revelado pouco efetiva na eliminação de biofilmes e é associada a um alto risco de contaminação das águas. Portanto, é necessário que se busquem abordagens mais eficientes e específicas em relação ao controle microbiológico. O uso de vírus bacteriófagos vem ao encontro dessas necessidades, pois eles, após se multiplicarem, geralmente provocam a lise celular, liberando novas partículas virais e evitando que a bactéria se prolifere. Diante disso, este estudo propõe estabelecer um método para a concentração e a determinação da eficiência de recuperação de bacteriófagos de BRS presentes em água de reator oriunda de poços de petróleo. As amostras foram coletadas de dois reatores operados em batelada alimentada e que simulam um poço de petróleo. As amostras de água de reator foram primeiramente clarificadas, os vírus eluídos desse sedimento e, em seguida, concentrados por ultracentrifugação. O concentrado viral foi então purificado com Vertrel XF. Ensaios de semeadura experimental de miofago P1 nas amostras de água do reator revelaram taxa de recuperação viral de 27,7%, contra ao 16% obtidos com outros protocolos.

ABSTRACT The control of microbial growth is an increasing challenge in the production of oil and gas, since the presence of certain bacteria has economic and environmental negative impacts. Sulphate reducing bacteria are particularly problematic, since they are responsible for the biological corrosion associated with the production of hydrogen sulfide, an effect known as souring. The main form of control is the use of biocides; however, this strategy, in addition to requiring continuous application, has proven to be ineffective in the elimination of biofilms and is associated with a high risk of water contamination. Therefore, it is necessary to seek more efficient and specific approaches to microbiological control. The use of bacteriophage viruses meets these needs, because after they multiply, they usually cause cell lysis, releasing new viral particles and preventing the bacteria from proliferating. In view of this, this study proposes to establish a method for the concentration and detection of bacteriophages of Sulphate Reducing Bacteria present in reactor water from oil wells. The samples were collected from two reactors, operated in a batch fed to simulate an oil well. The reactor water samples were first clarified, viruses adsorbed to sediment were eluted and then concentrated by ultracentrifugation. The viral concentrate was then purified with Vertrel-XF. Experimental seeding of P1 myophage in water samples from the reactor revealed a viral recovery rate of 27.7%, compared to the 16% obtained by use of other protocols.

Tetracyclines lead to ammonium accumulation during nitrification process.

The effect of tetracyclines used for swine food-production (tetracycline and oxytetracycline) on enriched nitrifying bacteria cultures over time was investigated in this study. Short-term exposure assays were performed in different concentrations of each antibiotic, using ammonia oxidizing bacteria (AOB) culture and nitrifying bacteria. The results pointed out a higher inhibitory effect of tetracycline on both bacterial communities. The AOB was more sensitive to antibiotic exposure when compared to the nitrifying culture. Although high antibiotic concentrations were applied, the half maximal inhibitory concentration (IC50) was achieved only for the AOB culture exposed to tetracycline at a concentration of 273 mg L-1. Nonetheless, the long-term exposure assay demonstrated a reduction of the tetracycline inhibition effect against AOB. The exposure to 100 mg L-1 of tetracycline (TC) did not show relevant influence over ammonium conversion efficiency; however, at 128 mg L-1 of TC, the efficiency decreased from 94% to 72%. Further investigation revealed that TC reduced the final effluent quality due to the development of a resistance mechanism by AOB culture against this antibiotic. This mechanism involves increasing the excretion of extracellular polymeric substances (EPS) and soluble microbial products (SMP), which probably increases BOD, and reduces ammonia consumption by the bacterial culture.

Potential of enzymatic process as an innovative technology to remove anticancer drugs in wastewater.

Anticancer drugs are a class of pharmaceutical compounds that have been found in hospital, domestic, and industrial wastewaters and also in surface waters. They have been showing recalcitrance to conventional wastewater treatment technologies and present a potential risk to environment and human health, since they exhibit cytotoxic, teratogenic, and carcinogenic among other effects in higher organisms, even at low concentrations. The presence of these compounds in the environment is a recent challenge for wastewater treatment and some alternative strategies to remove them were already studied, such as white-rot fungi (WRF) technologies. Despite promising results, processes involving fungi are complex, have high reaction times, and require nutrient addition for fungus growth and maintenance. Due to this potential, strategies to make the technology feasible were studied, such as the possibility for direct application of enzymes secreted by WRF. Enzymatic processes were studied in the removal of other pharmaceuticals such as antibiotics, anti-inflammatory, and steroid hormones; however, to the best of our knowledge, there is a gap on literature about their direct action on anticancer drugs.

Residual sugar from microalgae biomass harvested from phycoremediation of swine wastewater digestate.

The present study assessed the carbohydrate and sugar production from Chlorella spp. biomass harvested from a field scale reactor simulating phycoremediation of swine wastewater. The microalgae biomass was mainly composed by (%): carbohydrates (41 ± 0.4), proteins (50 ± 0.4), and lipids (1.3 ± 0.5). The residual sugar present in the biomass was extracted via acid hydrolysis. Among different concentrations of sulfuric acid tested (i.e., 47, 94, 188, 281 and 563 mM), significantly higher sugar content was obtained with 188 mM (0.496 g-sugar g-1 microalgae-DW). The concentration of sugar present in the microalgae did not differ significantly between the biomasses harvested by either centrifugation or coagulation-flocculation. Two commercially available strains of yeast (i.e., Saccharomyces cerevisiae and S. cerevisiae chardonnay) were tested for their capability to ferment sugar from lyophilized microalgae biomass. S. cerevisiae chardonnay showed a significantly faster consumption of sugar during the exponential growth phase. Both strains of yeast were capable of consuming most of the sugar added ≅ 8 g L-1 within 24 h. Overall, the results suggest that carbohydrate-rich microalgae biomass obtained from the phycoremediation of swine wastewaters can play an important role in green design for industries seeking alternative sources of feedstock rich in sugar.

Effects of Nitrogen and Phosphorus on Biochemical Composition of Microalgae Polyculture Harvested from Phycoremediation of Piggery Wastewater Digestate.

The effects of nitrogen (N) and/or phosphorus (P) starvation on the biochemical composition of native microalgae Chlorella spp. polyculture obtained from the phycoremediation of swine wastewaters were investigated. Microalgae-specific growth rate of 1.2 day(-1) was achieved (30.3 mg L(-1) day(-1)). PO4 (-2) and NH3 were completely removed from swine digestate effluent after 3 and 11 days, respectively. Microalgae harvested immediately after nutrient removal showed high protein (56-59 %) and carbohydrate (25-34 %) but low lipid (1.8-3 %) contents. Depletion of N or P alone stimulated carbohydrate production at the expenses of proteins. Significant lipid accumulation from 3 % ± 0.5 to 16.3 % ± 0.8 was reached only after 25 days following N and P starvation as demonstrated by Nile red-stained cells. Regarding to the effects of harvesting methods on cellular biochemical composition, circumstantial evidences indicate that coagulation-flocculation with tannin may lead to lower protein and lipid amounts but increased carbohydrate content (p < 0.01) as compared to centrifugation.

Enhancement of nutrient removal from swine wastewater digestate coupled to biogas purification by microalgae Scenedesmus spp.

This work investigated the effects of swine wastewater-derived biogas on microalgae biomass production and nutrient removal rates from piggery wastewater concomitantly with biogas filtration. Photobioreactors with dominant Scenedesmus spp. were prepared using non-sterile digestate and exposed to different photoperiods. In the presence of biogas and autotrophic conditions microalgae yield of 1.1±0.2 g L(-1) (growth rate of 141.8±3.5 mg L(-1) d(-1)) was obtained leading to faster N-NH3 and P-PO4(3-) assimilation rate of 21.2±1.2 and 3.5±2.5 mg L(-1) d(-1), respectively. H2S up to 3000 ppmv was not inhibitory and completely removed. Maximum CO2 assimilation of 219±4.8 mg L(-1) d(-1) was achieved. Biological consumption of CH4 up to 18% v/v was verified. O2 up to 22% v/v was controlled by adding acetate to exacerbate oxygen demand by microorganisms. Microalgae-based wastewater treatment coupled to biogas purification accelerates nutrient removal concomitantly producing valuable biomass and biomethane.

Drinking water decontamination by biological denitrification using fresh bamboo as inoculum source.

Groundwater contamination is becoming a serious problem in many Brazilian regions. European countries started to deal with this issue in the 1980s, mainly caused by the extensive usage of nitrogenous fertilizers and the absence of domestic wastewater treatment. Due to its high solubility, nitrate readily passes through the soil and reaches the aquifer. Thereafter, this ion moves, following groundwater flow, and can be found several kilometers from the area where the pollution occurred. Concern about nitrate contamination is due to the link found between this contaminant and various human health diseases, such as methemoglobin and cancer. Studies carried out in France enabled the design and implementation of several biological denitrification plants throughout the country, in order to remove nitrate from its contaminated groundwater. Heterotrophic denitrification facilities shown to be adequate to treat high water flows with satisfactory nitrate removal efficiency, especially when static media supports are employed. The objective of this research was to evaluate the existence of denitrifying microorganisms in bamboo (Bambusa tuldóides) and verify the feasibility of their use to inoculate a pilot-scale fixed-bed bioreactor. The support material selected to fill the bioreactor bed was commercial polypropylene Pall rings, since such support has a high porosity associated with a wide superficial area. The bioreactor was able to produce and retain a large amount of cells. Using ethanol as carbon source, nitrate (N-NO3(-)) removal efficiency of the bioreactor stood around 80 % for a maximum nitrogen loading rate of approximately 6.5 mg N-NO3 (-) L(-1) h(-1).

Study of banana (Musa aaa Cavendish cv Nanica) trigger ripening for small scale process

The present work focuse on the impact of O2, CO2 and ethylene concentrations on ripening rate control of bananas as a contribution for the development of domestic equipments that could allow the user to drive the fruit shelf live. It represented the adjustment of metabolic activity rates in order to manage the maturity process. Ripening variables such as ethylene and CO2 concentrations and temperature were adjusted to accelerate or slow down the process, while the maturity degree was monitored through the physical and chemical parameters and sensorial analysis. Therefore, the objective of this work was to evaluate the influence of these parameters to manage the banana ripening. The optimum temperature was at 25 ºC of storage. The presence of oxygen, CO2 withdraws and ethylene injection were relevant for the ripening process. The "ready-to-eat" quality was achieved in 6 days in confined system. The use of ethylene as trigger was adequate to accelerate the ripening process with advantages in fruit color.

O presente trabalho foca no impacto da concentração de O2, CO2 e etileno no controle da taxa de amadurecimento de bananas, como contribuição para o desenvolvimento de equipamentos domésticos que permitam o controle pelo próprio usuário da vida de prateleira de frutas. Isto representa o ajuste das atividades metabólicas para garantir o controle do amadurecimento. Variáveis como concentração de etileno e CO2 e temperatura foram ajustadas para acelerar ou reduzir o processo, enquanto que o grau de maturação foi monitorado através de parâmetros físico-químicos e sensoriais. Desta forma, o objetivo deste trabalho foi avaliar a influência destes parâmetros para controlar o amadurecimento de banana. A temperatura ótima de amadurecimento foi 25ºC. A presença de O2, a retirada de CO2 e a injeção de etileno foram relevantes no processo. A qualidade "pronto-para-consumo" foi obtida em 6 dias em sistema confinado. O uso de etileno como gatilho é adequado para acelerar o amadurecimento, com vantagens para a cor do produto.