Fabrication of PVA-Silver nanoparticle composite film for elimination of microbial contaminant from effluent

The effluent contains many harmful microbes which should be eliminated before it is discharged into a water body. Silver nanoparticles (AgNPs) being high-quality significance and have a great impact on this research field as it inhibits microbial proliferation and infection. Therefore, it may use for Bioremediation purposes, our laboratory is fascinated by the production of polymer matrix entrapment silver nanoparticles for in situ bio-remediation purposes. The AgNPs was prepared from sawdust by decoction method. The yellowish solution turns into dark brown colour indicating the formation of AgNPs. A sharp SPR (Surface Plasmon Resonance) band formation in UV-vis spectroscopy scan establishes the formation and stability of silver nanoparticles in an aqueous solution. SEM microphotograph indicated roughly spheroidal structure with (63±3) nm average diameters of newly synthesized AgNp. Polyvinyl alcohol (PVA) is eco-friendly and non-toxic to the environment was chosen for the preparation of polymeric matrix. The non-toxic concentration (1 ?g/mL) of AgNp was dispersed into PVA solution followed by cross-linked with maleic acid. PVA- maleic acid is cross-linked by the formation of an ester bond, whereas silver nanoparticles physically entrap into the cross-linked matrix. The silver nanoparticles were released from the matrix nearly after 10 min of swelling of the composite film. In a microbial assay using E. coli agar medium, PVA-AgNp composite film shows the significant killing of microorganisms. Microbial elimination is measured indirectly by pH measurement and dissolved oxygen concentration measurement of the effluent in situ against RO- water, taken as control. The dissolved oxygen concentration from RO water and effluent water was measured on Day “0” followed by treatment and incubation at the BOD chamber. The treatment with PVA-AgNp composite film reduced the BOD Level and increase dissolved oxygen level simultaneously increasing the quality of water.

Comparación entre manguera difusora y tubería PVC para la aireación en la producción de Litopenaeus vannamei con biofloc a diferentes niveles de proteína / Comparison between diffuser hose and PVC pipe for aeration in production of Litopenaeus vannamei with biofloc at different protein levels / Comparação entre mangueira difusora e tubo de PVC para aeração na produção de Litopenaeus vannamei com bioflocos em diferentes níveis de proteína

Resumen Al implementar la tecnología biofloc, la aireación es uno de los problemas para obtener buenos resultados productivos, por tanto, el objetivo fue comparar la eficiencia de la tubería PVC y manguera difusora, como sistema de aireación en cultivos con biofloc y dos niveles de proteína. Se aplicó un diseño completamente al azar con arreglo factorial con cuatro tratamientos, tres réplicas cada uno, con los dos sistemas de aireación y dos niveles de proteína bruta (PB) en el alimento balanceado en producciones de camarón Litopenaeus vannamei, distribuidos de la siguiente manera: para el tratamiento uno (T1) se utilizó tubería PVC y 28% de PB; para T2, manguera difusora y 22% de PB; para T3, manguera difusora y 28% de PB; para T4, tubería PVC y 22% de PB, todos producidos con biofloc. Las variables peso, talla, alimentación y el factor tratamiento influencian significativamente en la concentración de oxígeno disuelto (OD, mg/l) en la tarde, T1 es el que mejor se comportó en la tarde y T3 en la mañana. El tratamiento con menor sedimentación a los 20 y 30 minutos en la prueba de cono Imhoff fue T4 y el de mayor sedimentación fue el T3. No hubo diferencia significativa en la supervivencia entre tratamientos, los niveles de proteína sí influencian en las concentraciones del OD. El uso de tubería PVC aporta en el incremento de peso y de la concentración de OD en relación al sistema de manguera difusora con el 22% y 28% de proteína bruta.

Abstract When implementing biofloc technology, aeration is one of the problems to obtain good productive results, therefore, the objective was to compare the efficiency of PVC pipe and diffuser hose, as an aeration system in crops with biofloc and two levels of protein. A completely randomized design with factorial arrangement was applied with four treatments with three replicates each, with the two aeration systems and with two levels of crude protein (CP) in the balanced feed in Litopenaeus vannamei shrimp productions, distributors of the following way, for treatment one (T1) PVC pipes and 28% PB were achieved; for T2, diffuser hose and 22% PB; for T3, diffuser hose and 28% PB; for T4, PVC pipe and 22% PB, produced with biofloc. The variables weight, height, feeding and the treatment factor significantly influence the concentration of dissolved oxygen (DO, mg/l) in the afternoon. T1 is the one that behaves best in the afternoon and T3 in the morning, the treatment with the lowest sedimentation at 20 and 30 minutes in the Imhoff cone test was T4 and the highest sedimentation was T3. There was no significant difference in survival between treatments, protein levels did influence OD concentrations. The use of PVC pipe provides an increase in weight and DO concentration in relation to the diffuser hose system with 22% and 28% crude protein.

Resumo Ao implementar a tecnologia de bioflocos, a aeração é um dos problemas para obter bons resultados produtivos, portanto, objetivou-se comparar a eficiência do tubo de PVC e da mangueira difusora, como sistema de aeração em lavouras com bioflocos e dois níveis de proteína. Foi aplicado um delineamento inteiramente casualizado com arranjo fatorial com quatro tratamentos com três repetições cada, com os dois sistemas de aeração e com dois níveis de proteína bruta (PB) na ração balanceada em produções de camarão Litopenaeus vannamei, distribuidores da seguinte forma, para tratamento com tubo de PVC (T1) e PB de 28%; para T2, mangueira difusora e PB 22%; para T3, mangueira difusora e PB 28%; para T4, tubo de PVC e PB 22%, produzido com biofloco. As variáveis peso, altura, alimentação e o fator de tratamento influenciam significativamente na concentração de oxigênio dissolvido (OD, mg/l) no período da tarde, T1 é o que apresenta melhor desempenho no período da tarde e T3 no período da manhã, tratamento com menor sedimentação aos 20 e 30 minutos no teste do cone de Imhoff foi T4 e a maior sedimentação foi T3. Não houve diferença significativa na sobrevivência entre os tratamentos, os níveis de proteína influenciaram as concentrações de DO. A utilização de tubo de PVC proporciona um aumento de peso e concentração de OD em relação ao sistema de mangueiras difusoras com 22% e 28% de proteína bruta.

Biosynthesis of lipase by Burkholderia cenocepacia ST8 using waste cooking oil as feedstock

Aims@#Every year, an estimated 25 million tons of waste oil are produced worldwide, and the generation of waste oil is one of the biggest global environmental problems. The incorporation of oil as a substrate for lipase production has been studied and shown to have a positive impact on its production. Burkholderia sp. is one of the major lipase-producing bacteria with their ability in bioremediation of oil-contaminated soil. This study aims to compare the production of lipase by Burkholderia cenocepacia ST8 using waste cooking oil and unused cooking oil as feedstock.@*Methodology and results@#The effect of different types of waste cooking oil (sunflower oil and palm oil) and concentration (1-3%) of waste cooking oil, agitation speed (100-400 rpm) and initial dissolved oxygen concentration (10-50%) on lipase production by B. cenocepacia ST8 under batch fermentation mode were investigated. The major fatty acids of which had been consumed were determined using gas chromatography. Results showed that 2% (v/v) of single used sunflower cooking oil produced the highest lipase activity of 138.86 U/mL with a productivity of 2.10 U/mL/h; agitation speed of 300 rpm produced the highest lipase activity of 183.56 U/mL with a productivity of 3.06 U/mL/h while 30% initial concentration of dissolved oxygen produced a lipase activity of 176.45 U/mL with a productivity of 2.94 U/mL/h. Oleic acid and linoleic acid were found to be the most consumed by B. cenocepacia ST8 among other fatty acids. @*Conclusion, significance and impact of study@#This study shows that 2% (v/v) single used sunflower cooking oil was the better type and optimum concentration of carbon source for the production of lipase by the fermentation of B. cenocepacia under 300 rpm and 30% initial concentration dissolved oxygen. The incorporation of 2% (v/v) single used sunflower cooking oil may be a great alternative to reduce the cost for the production of lipase as well as reducing the amount of waste oil generation.

Sensibilidad de Elmidae (Insecta: Coleoptera) a la perturbación delhábitat y la calidad fisicoquímica del agua en ambientes lóticos de los Andes colombianos

Introducción: Los élmidos son organismos acuáticos frecuentemente asociados con aguas corrientes, limpias y bien oxigenadas. Sin embargo, algunos géneros de esta familia también se encuentran en aguas moderadamente contaminadas, lo cual puede ser un reflejo de su tolerancia a algún grado de deterioro del hábitat. Objetivo: Determinar la sensibilidad o tolerancia a la contaminación y los principales factores que influyen en la presencia y composición taxonómica de Elmidae en microcuencas colombianas. Métodos: Se muestrearon 60 localidades entre 450 y 4 000 msnm, principalmente en la región Andina, los muestreos se realizaron entre 2002 y 2013 con 70 eventos no simultáneos en cinco zonas. En un transecto de 100 m, se registraron datos físico-químicos, bacteriológicos y de calidad del hábitat (ICH), se hicieron arrastres con red D y Surber, y se determinaron los géneros de Elmidae. Se realizó un Análisis de Correspondencia Canónica entre las variables ambientales y la composición de Elmidae, las relaciones significativas se confirmaron con una prueba de Mantel y se realizó una prueba Mantel parcial incluyendo la ubicación geográfica como tercera matriz. Se calcularon los Índices de Contaminación ICOs y de Calidad de Agua ICA-FSN y se definieron intervalos de aparición de géneros para variables ambientales e índices. Para obtener las significancias estadísticas, se realizaron Análisis de Varianza ANOVA, pruebas t de Student y Modelos Lineales Generalizados MLG con distribución Poisson. Resultados: Se encontraron 22 géneros y 15 especies que representan el 78.5 % de la riqueza nacional conocida. Tyletelmis y las especies Gyrelmis pusio, G. simplex, Xenelmis sandersoni, X. leechi, X. granatoides y X. teres, corresponden a nuevos registros para Colombia. Los principales factores que influyeron en la riqueza y composición de Elmidae fueron: saturación de oxígeno, deposición de sedimentos,sólidossuspendidos, disponibilidad de sustratos, altura, ubicación geográfica, conductividad, alcalinidad, turbidez, nitratos, nitrógeno amoniacal y fósforo. Conclusiones: Los géneros de Elmidae difieren en su tolerancia a la contaminación y, por lo tanto, no pueden ser clasificados bajo el mismo grado de sensibilidad. Esta información puede ser útil para la clasificación de los géneros de Elmidae de acuerdo con su sensibilidad a los cambios en las condiciones ambientales y la degradación del hábitat, lo cual será un primer paso para la incorporación en un índice biótico de calidad de agua adaptado para las condiciones locales o el ajuste de los índices preexistentes.

Introduction: Riffle beetles are aquatic organisms frequently associated with well-oxygenated clean running water. However, some genera of Elmidae can also be found in moderately polluted streams proving their tolerance to certain degree of habitat degradation. Objective: To determine the pollution sensitivity and tolerance of each genus of Elmidae in Colombian streams, and the main factors that influence its presence and taxonomic composition. Methods: Sixty locations between 450 and 4 000 m.a.s.l. were sampled, mainly in the Andean Region of Colombia. Collection was carried out between 2002 and 2013 with 70 non-simultaneous events in five zones. In a 100 m transect, physicochemical, bacteriological and environmental quality information was recorded. Samples of Elmidae were collected with D and Surber nets and determined taxonomically. A Canonical Correspondence Analysis of the Elmidae assembly and environmental data was carried out, significant relationships were confirmed with a Mantel test and a partial Mantel test was carried out including the geographic location as the third matrix. The Pollution (ICO) and Water Quality (ICA-NSF) Indices were calculated, and the occurrence intervals by genus were plotted for environmental variables and indices. Analysis of Variance ANOVA, Student's t-test and Generalized Linear Models GLM with Poisson distribution, were calculated in order to establish the significance of the results. Results: Twenty-two genera and 15 species were found in the evaluated streams; which represent 78.5 % of known national generic richness. Tyletelmis and the species Gyrelmispusio, G. simplex, Xenelmis sandersoni, X. leechi, X. granatoides and X. teres correspond to new records for Colombia. The most important factors influencing the presence and composition of Elmidae were: oxygen saturation, sediment deposition, suspended solids, epifaunal substrate availability, altitude and geographic location, conductivity, turbidity, alkalinity, nitrates, ammonia nitrogen and phosphorus. Conclusions: The genera of Elmidae differ in their tolerance to contamination and therefore cannot be classified in the same degree of sensitivity. This information can be useful for classifying genera of Elmidae according to their sensitivity to changes in environmental conditions and habitat degradation, which will be a first step to incorporate Elmidae in water quality biotic indices adapted to local conditions or to adjust preexisting indices.

Influence of Oxygen Supply on Growth and Laccases Production by Pleurotus sajor-caju PS-2001 in Submerged Process

Abstract (1) Background: Oxygen supply is an important parameter to be considered in submerged cultures. This study evaluated the influence of different conditions for dissolved oxygen (DO) concentration on laccases activities and growth of Pleurotus sajor-caju PS-2001 in submerged process in stirred-tank bioreactor. (2) Methods: Initially, three different conditions were tested: uncontrolled DO and minimum levels of 30% and 80% of saturation, with the pH controlled between 4.5 and 7.0. (3) Results: Best results were observed at 30% DO (26 U mL-1 of laccases at 96 h), whereas higher mycelial biomass was observed at 30% and 80% DO (above 4.5 g L-1). Four different conditions of DO (uncontrolled, 10%, 30% and 50% of saturation) were tested at pH 6.5, with higher laccases activity (80 U mL-1 at 66 h) and lower mycelial growth (1.36 g L-1 at 90 h) being achieved with DO of 30%. In this test, the highest values for volumetric productivity and specific yield factor were determined. Under the different pH conditions tested, the production of laccases is favoured at DO concentration of 30% of saturation, while superior DO levels favours fungal growth. (4) Conclusion: The results indicate that dissolved oxygen concentration is a critical factor for the culture of P. sajor-caju PS-2001 and has important effects not only on laccases production but also on fungal growth.

Efecto de la temperatura y salinidad en la mortalidad de adultos de Capitella sp. (Polychaeta: Capitellidae) en el laboratorio

Introducción: El poliqueto marino y estuarino Capitella capitata es una especie endobentónica y considerado cosmopolita. El cultivo en el laboratorio de este complejo de especies permite su utilización como especie para pruebas en bioensayos de toxicidad, así como para estudios de reproducción. Objetivo: El presente trabajo pretende observar la mortalidad en dos rangos de temperatura y salinidad como condiciones óptimas para cultivar Capitella sp. en el laboratorio. Metodología: La recolección de especímenes se realizó en la provincia de Callao, Perú en noviembre de 2016 y enero de 2017. Se realizó un bioensayo para observar la mortalidad bajo diferentes intervalos de temperatura y salinidad: tratamiento 1 con los organismos recolectados en enero y con intervalos de temperatura (20-22° C) y salinidad (30-32 ups) similares a los registrados en el campo; tratamiento 2, con los organismos recolectados en noviembre y con intervalos de temperatura (24-26 °C) y salinidad (32 a 34 ups) seleccionados de acuerdo con datos de la bibliografía. Resultados: El aumento de la temperatura y el tiempo influyeron significativamente en la mortalidad de los organismos de ambos tratamientos (Correlación de Spearman), mientras que la salinidad y el oxígeno no presentaron correlación significativa con la mortalidad. Se observó efecto significativo del tiempo en la mortalidad, con más muertes a partir de la segunda mitad del experimento. Los organismos del tratamiento 1, con condiciones ambientales similares a las de su hábitat sobrevivieron más que los del tratamiento 2. Estos resultados también se observaron en la variabilidad porcentual en el coeficiente de variación entre los días por tratamiento. No se observaron diferencias entre los tratamientos al comparar las curvas acumuladas de mortalidad. Conclusión: Los intervalos óptimos para el cultivo en el laboratorio de Capitella sp. son los de su hábitat. Las posibles causas de la mortalidad observada se atribuyen, además de la temperatura y el tiempo, a la edad, estado de salud o de madurez de los organismos y, probablemente, la mala calidad del agua utilizada en el bioensayo.

Introduction: The marine-estuarine polychaete Capitella capitata is an endobenthic species which is considered as a cosmopolitan organism. The laboratory culture of this species-complex allows its use as a test species in toxicity bioassays and reproduction studies. Objective: It is pretended to observe mortality under two temperature and salinity ranges as optimal conditions to culture Capitella sp. in the laboratory. Metodology: Polychaetes were collected in Callao Province, Peru, during November 2016 and January 2017. A bioassay was carried out to observe mortality under different temperature and salinity ranges using two treatments: treatment 1, with polychaetes collected during January 2017, which were subjected to conditions similar to the collection site (temperature: 20-22° C, salinity: 30-32 psu); treatment 2, with the organisms collected during November 2016, with temperature (24-26 °C) and salinity (32 a 34 ups) selected according to bibliographic data. Results: The increase of temperature and time significantly influenced polychaete mortality of the two treatments (Spearman correlation), while salinity and dissolved oxygen did not show correlation with mortality. Significant effect of time was observed on mortality, with more deaths from the second half of the experiment. Organisms from treatment 1, with environmental conditions similar to those of their habitat survived more than worms from treatment 2. These results were also observed in the porcentual varibility of the variation coefficient among days in each treatment. The cumulative mortality curve did not show differences between treatments. Conclusion: The optimal ranges to culture Capitella sp. in the laboratory are those from their habitat. In addition to temperature and time, other posible causes of the observed mortality can be worms age, health or maturity, and probably the bad quality of the water used during the bioassay.

Estimativa do coeficiente de transferência de massa de oxigênio com o uso da técnica das sondas solúveis flutuantes: um estudo de laboratório / Estimating the mass transfer coefficient using the floating soluble probes technique: a laboratory study

RESUMO A sonda solúvel flutuante (SSF) é considerada uma técnica de baixo custo para a estimativa do coeficiente de transferência de massa de oxigênio na interface ar-água (KL). Essa técnica se baseia no fato que KL pode ser correlacionado com a velocidade de dissolução do sólido solúvel (VS), uma vez que ambos os parâmetros são influenciados pela turbulência na superfície do escoamento. O objetivo do presente estudo consistiu em calibrar uma nova configuração da SSF. Experimentos de reaeração superficial e dissolução da sonda solúvel foram realizados em dois aparatos experimentais (tanque agitado por jatos e um canal hidráulico circular), de modo que diferentes níveis de turbulência foram produzidos. Os resultados mostraram forte correlação entre KL e VS; r = 0,953 para experimentos realizados no tanque e r = 0,895 para experimentos realizados no canal. A nova configuração da SSF forneceu razão KL.VS -1 crescente em função do número de Reynolds, indicando que a relação entre os dois parâmetros é dependente da turbulência. Conclui-se que a técnica SSF possui elevado potencial. Entretanto, para que se possa atribuir utilidade definitiva, verificações em cursos d'água naturais e em equipamentos com estrutura de turbulência diferentes das utilizadas neste estudo ainda são necessárias.

ABSTRACT The floating solid soluble probe is a low-cost technique for estimating the mass transfer coefficient of oxygen in the air-water interface (Ka). This technique is based on the fact that Ka can be correlated with the dissolution rate of a soluble solid (VS), since both parameters are influenced by the surface turbulence. The aim of this study was to calibrate a new floating solid soluble probe configuration. Surface reaeration experiments and dissolution of the soluble solid probe were performed in two experimental apparatus (jets stirred tank and a circular hydraulic channel), in which different levels of turbulence were produced. The results showed a strong correlation between Ka and VS, r = 0.953 for experiments performed on the tank and r = 0.895 for experiments performed on the channel. The new floating solid soluble probe configuration provided an increasing Ka.VS -1 ratio as a function of the Reynolds number, indicating that the relationship between these two parameters depends on the surface turbulence. We conclude that the floating solid soluble probe technique has a high potential, despite calibrations in natural water bodies and equipment with a turbulence structure different from those used in this study are still necessary to definitive utility.

Factors influencing the accuracy of the denitrifier method for determining the oxygen isotopic composition of nitrate / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The denitrifier method is widely used as a novel pretreatment method for the determination of nitrogen and oxygen isotope ratios as it can provide quantitative and high-sensitivity measurements. Nevertheless, the method is limited by relatively low measurement accuracy for δ18O. In this study, we analyzed the factors influencing the accuracy of δ18O determination, and then systematically investigated the effects of dissolved oxygen concentrations and nitrate sample sizes on estimates of the δ15N and δ18O of nitrate reference materials. The δ18O contraction ratio was used to represent the relationship between the measured difference and true difference between two reference materials. We obtained the following main results: (1) a gas-liquid ratio of 3:10 (v/v) in ordinary triangular flasks and a shaking speed of 120 r/min produced an optimal range (1.9 to 2.6 mg/L) in the concentration of dissolved oxygen for accurately determining δ18O, and (2) the δ18O contraction ratio decreased as nitrate sample size decreased within a certain range (1.0 to 0.1 μmol). Our results suggested that δ18O contraction is influenced mainly by dissolved oxygen concentrations in pure culture, and provided a model for improving the accuracy of oxygen isotope analysis.

Factors influencing the accuracy of the denitrifier method for determining the oxygen isotopic composition of nitrate / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The denitrifier method is widely used as a novel pretreatment method for the determination of nitrogen and oxygen isotope ratios as it can provide quantitative and high-sensitivity measurements. Nevertheless, the method is limited by relatively low measurement accuracy for δ18O. In this study, we analyzed the factors influencing the accuracy of δ18O determination, and then systematically investigated the effects of dissolved oxygen concentrations and nitrate sample sizes on estimates of the δ15N and δ18O of nitrate reference materials. The δ18O contraction ratio was used to represent the relationship between the measured difference and true difference between two reference materials. We obtained the following main results: (1) a gas-liquid ratio of 3:10 (v/v) in ordinary triangular flasks and a shaking speed of 120 r/min produced an optimal range (1.9 to 2.6 mg/L) in the concentration of dissolved oxygen for accurately determining δ18O, and (2) the δ18O contraction ratio decreased as nitrate sample size decreased within a certain range (1.0 to 0.1 μmol). Our results suggested that δ18O contraction is influenced mainly by dissolved oxygen concentrations in pure culture, and provided a model for improving the accuracy of oxygen isotope analysis.

Produção de L-asparaginase pela levedura Leucosporidium muscorum CRM 1648 isolada de sedimento marinho coletado na Península Antártica / L-asparaginase production by the yeast Leucosporidium muscorum CRM 1648 isolated from marine sediments collected in Antarctic Peninsula

L-asparaginase (L-ASNase) é uma enzima com propriedades interessantes para a indústria médica, farmacêutica e de alimentos, que tem recebido atenção especial, inclusive no Brasil, por fazer parte do protocolo de tratamento de distúrbios linfoproliferativos, como a leucemia linfoblástica aguda (LLA). No mercado desde a década de 1970, as enzimas de origem bacteriana enfrentam algumas limitações por provocarem reações adversas graves em quase 80% dos pacientes em tratamento. Nesse contexto, L-ASNases provenientes de leveduras se destacam como alternativa, por serem mais próximas às congêneres humanas. A Antártica ainda é um ambiente pouco explorado, com grande diversidade de microrganismos com potencial para a produção de moléculas biológicas de interesse industrial. Nesse contexto, 150 leveduras isoladas de amostras de sedimento marinho coletadas na Península Antártica como parte do projeto MICROSFERA (PROANTAR/CNPq) foram avaliadas para a produção de L-ASNase. A triagem resultou em 9 isolados produtores, dos quais 7 pertencem ao gênero Leucosporidium. A linhagem L. muscorum CRM 1648 foi a que produziu mais enzima (540 U.L-1), com maior produtividade (5,6 U.L-1.h-1) e, por isso, foi alvo deste estudo. A análise univariada de fontes de carbono e nitrogênio indicou maior crescimento desse microrganismo e produção de L-ASNase em meio CD com extrato de levedura, prolina e sacarose. Ureia, cloreto de amônio e sulfato de amônio resultaram em baixa ou nenhuma produção da enzima, sugerindo que a metabolização de fontes de nitrogênio por essa linhagem está sob a influência do fenômeno de repressão catabólica pelo nitrogênio (RCN). Dois delineamentos experimentais do tipo fatorial completo resultaram em um aumento de 10 vezes na produção e produtividade da enzima (4582,5 U.L-1 e 63,6 U.L-1.h-1, respectivamente). A análise univariada da concentração inicial de inóculo (X0), pH inicial do meio, temperatura e adição de água do mar mostrou que a melhor condição para a produção foi: pH = 5,5 ou 6,5, cultivo a 15°C com adição de água do mar (25-50% m/v). A variável X0 não foi significativa nas concentrações avaliadas. Cultivos em biorreator (batelada) foram conduzidos em quatro diferentes níveis de oxigênio dissolvido (OD): (1) OD não controlado e abaixo de 20%, (2) OD não controlado e acima de 20%, (3) OD controlado em 80% e (4) OD controlado em 20%. Os resultados mostraram que OD é fator limitante para o crescimento de L. muscorum CRM 1648 e produção de L-ASNase por essa levedura e deve ser mantido acima de 35% para maior produção da enzima.Neste trabalho, a composição do meio e condições de cultivo foram estabelecidas para favorecer a produção de uma nova L-ASNase livre de atividade glutaminásica por levedura adaptada ao frio, abrindo espaço para novos estudos acerca de seu potencial antileucêmico e possível uso como alternativa às enzimas já existentes no mercado no tratamento de LLA

L-asparaginase (L-ASNase) is an enzyme with interesting properties for medical, pharmaceutical and food industry, which has received special consideration, especially in Brazil, for being part of lymphoproliferative disorders treatment, such as acute lymphoblastic leukemia (ALL). Bacterial enzymes are on the market since the 1970s and face some limitations related to theirserious adverse reactions that reach almost 80% of all patients in treatment. In this context, L-ASNases from yeasts are highlighted as important alternative to bacterial enzymes, due to the closerphylogeny to human congeners. Antarctic environment has much to be explored, with a vast diversity of microorganisms with potential to produce biomolecules with industrial interest. A total of 150 yeasts isolated from Antarctic marine sediments as part of MICROSFERA project (PROANTAR/CNPq) were evaluated for L-ASNase production. The screening resulted in 9 producers, 7 species from the genus Leucosporidium. L. muscorum CRM 1648 was the strain that yielded the highest L-ASNase activity (540 U.L-1) and volumetric productivity (5.6 U.L-1.h-1). Carbon and Nitrogen sources were evaluated by a method of one-factor at a time (OFAT). From the gather results, sucrose, yeast extract and proline resulted in a maximal growth and highest enzyme production.The absence or low production of L-ASNase in medium with urea, ammonium chloride and ammonium sulfate suggests the presence of nitrogen catabolic repression (NCR). Carbon and nitrogen concentration were evaluated by full factorial design and yielded about ten times higher enzyme and volumetric productivity (4582.5 U.L-1 and 63.6 U.L-1.h-1, respectively). Initial inoculum concentration (X0), initial pH, temperature and concentration of seawater in the culture were evaluated by OFAT analysis and the best condition for L-ASNase production was: pH = 5.5 or 6.5, at 15 °C with addition of seawater (25-50 wt%). X0 was not considered a significant variable. Bioreactor assays (in batch regime) were performed in four different dissolved oxygen (DO) levels: (1) without DO control (DO remained under 20%), (2) without DO control (DO remained above 20%), (3) DO controlled at 80%, and (4) DO controlled at 20%.The results showed that DO is a key factor for growth of L. muscorum CRM 1648 and production of L-ASNase by this yeast and should be maintained above 35% for higher production of this enzyme.At this work, the medium and culture conditions were established to support the production of a novel glutaminase-free L-ASNase by a cold adapted yeast, opening a new path for further studies regarding its antileukemic potential and possible use as an alternative for ALL treatment

Hydrogen productivity of hydrogen-rich sticks increased by ammonium salt / 军事医学

Objective To measure the catalytic effect of ammonium salt solution on hydrogen production , and analyze the relationships between hydrogen concentration and oxygen concentration , oxidation-reduction potential , and pH in solution . Methods Magnesium and water reacted at different concentrations of NH4Cl, (NH4)2SO4, NH4HCO3,(NH4)2CO3, NaHCO3 and Na2SO3 in 40℃water bath for 0, 2, 4, 6, 8 and 10 hours, and above-mentioned four indicators of ammonium salt solution were measured after these reactions .Results The hydrogen concentration in the solution increased with the reaction time and the concentration of the ammonium salt solution .Oxygen concentration and oxidation-reduction potential decreased .The hydrogen concentration in the solution was significantly negatively correlated with the oxygen concentration (r=-0.984).pH increased with the hydrogen concentration in the ammonium salt solution .Conclusion The ammonium salt solution has a good catalytic effect on magnesium and water reaction .NH4Cl has the strongest catalytic effect under the same reaction conditions,followed by (NH4)2SO4, NH4HCO3 and (NH4)2CO3.Thus, this study can provide detailed data on hydrogen production in different solutions .

Measurement and analysis of dissolved oxygen in circulating water of electromagnetic extracorporeal shock wave lithotripter / 医疗卫生装备

Objective To measure the maximum dissolved oxygen concentration of the lithotripter to avoid visible air bubbles during lithotripsy.Methods The present situation of degassing of lithotripter and clinical requirements for water were introduced.Under the following conditions of 7.5 L water capacity,35 L/h water flow,ambient temperature 28 ℃,17 kV,60 shocks per minute,the lithotripter discharged 10 000 shocks continuously without a visible air bubble in the circulating water,then the dissolved oxygens in the water were measured by using AZ8403 dissolved oxygen meter.The maximum value of dissolved oxygen in the water was determined therefore without a visible air bubble.Results The visible tiny air bubble emerged during lithotripsy if the dissolved oxygen in the water was above 3.37 mg/L and ambient temperature was about 28 ℃;the bubble could be avoided if the dissolved oxygen was under 3.34 mg/L and ambient temperature was about 28 ℃.Conclusion The circulating water with dissolved oxygen concentration less [han 3 mig/L and about 28 ℃ ambient temnperature are recommended for the electromagnetic extracorporea] shock wave lithotripter during lithotripsy.

Biogeochemical validation of an interannual simulation of the ROMS-PISCES coupled model in the Southeast Pacific / Validación biogeoquímica de una simulación interanual del modelo acoplado ROMS-PISCES en el Pacífico Sudeste

Currently biogeochemical models are used to understand and quantify key biogeochemical processes in the ocean. The objective of the present study was to validate predictive ability of a regional configuration of the PISCES biogeochemical model on main biogeochemical variables in Humboldt Current Large Marine Ecosystem (HCLME). The statistical indicators used to evaluate the model were the bias, root-mean-square error, correlation coefficient and, graphically, the Taylor’s diagram. The results showed that the model reproduces the dynamics of the main biogeochemical variables (chlorophyll, dissolved oxygen and nutrients); in particular, the impact of El Niño 1997-1998 in the chlorophyll (decrease) and oxygen minimum zone depth (increase). However, it is necessary to carry out sensitivity studies of the PISCES model with different key parameters values to obtain a more accurate representation of the properties of the Ocean.

Los modelos biogeoquímicos en la actualidad son utilizados para entender y cuantificar los principales procesos biogeoquímicos que suceden en el océano. El objetivo del presente estudio es validar estadísticamente la habilidad predictiva de una simulación del modelo biogeoquímico PISCES en reproducir la dinámica de las principales variables biogeoquímicas del Ecosistema de la Corriente de Humboldt (ECH). Para evaluar el modelo se utilizaron indicadores estadísticos: sesgo, error de la raíz del cuadrado medio, coeficiente de correlación y gráficamente el diagrama de Taylor. Los resultados muestran que el modelo es capaz de reproducir la dinámica de las principales variables biogeoquímicas (clorofila, oxígeno disuelto y nutriente), captando bien el impacto que tiene El Niño 1997-1998 en la clorofila (disminución) y profundidad de la zona mínima de oxígeno (incremento). Es necesario llevar a cabo estudios de sensibilidad del modelo PISCES usando diferentes valores de los principales parámetros para obtener una mejor representación de las propiedades biogeoquímicas del océano.

Hypoxia increases susceptibility of Pacific white shrimp to whitespot syndrome virus (WSSV) / Condição de hipóxia aumenta a susceptibilidade do camarão branco do Pacíficoao vírus da mancha branca (WSSV)

The present study aimed to evaluate the mortality, reactive oxygen species production (ROS) and total hemocyte counts (THC) of the marine shrimp Litopenaeus vannamei infected with the white spot syndrome virus (WSSV) at three levels of oxygen saturation. For this, 360 shrimp (20±2g) were distributed in 24 tanks (60L), divided in two groups (infected and non-infected), which were subjected to 30, 60 and 100% of dissolved oxygen saturation (in quadruplicate). During 96 hours after infection, daily hemolymph samples were collected for hemato-immunological parameter evaluation (THC and ROS) and dead animals were removed and computed to assess cumulative mortality rates. In the infected group, animals subjected to 100% saturation showed higher ROS production (P<0.05) after 48 hours, while THC was significantly reduced (P<0.05), regardless of oxygen saturation. The hypoxia resulted in high mortality when compared to 100% saturation condition. In the uninfected group, no significant differences were observed in all evaluated parameters. Thus, the hypoxia condition increased the susceptibility of shrimp to the infection of WSSV, which may be partly related to the low ROS production showed by the animals subjected to 30% oxygen saturation.

O presente estudo teve por finalidade avaliar a mortalidade e a contagem total de hemócitos (CTH) e espécies reativas de oxigênio (EROs) de camarão Litopenaeus vannamei infectados com o vírus da mancha branca (WSSV) e submetidos a três níveis de saturação de oxigênio. Para tanto, 360 camarões (20±2g) foram distribuídos em 24 tanques (60L), divididos em dois grupos, infectados e não infectados e submetidos a 30, 60 e 100% de saturação de oxigênio (em quadruplicata). Após a infecção, diariamente foram coletadas amostras de hemolinfa dos animais para avaliação dos parâmetros hematoimunológicos (CTH e EROs) e foi estimada a mortalidade, por 96 horas. No grupo com infecção, os animais submetidos à saturação de 100% apresentaram um aumento na produção de EROs (P<0,05) após 48 horas, ao mesmo tempo em que a CTH demonstrou uma redução (P<0,05) independentemente da saturação do oxigênio, e a condição de hipóxia acarretou maiores mortalidades quando comparada à do grupo com 100% de saturação. No grupo sem infecção, não foram observadas diferenças significativas nos parâmetros avaliados nem mortalidade. Dessa forma, pode-se concluir que a hipóxia aumentou a susceptibilidade do camarão à infecção com o vírus da mancha branca, que pode estar, em parte, relacionada com a baixa contagem de hemócitos e produção de EROs observadas nos animais submetidos a essa condição.

Hydrogen-rich water prepared by hydrogen rods:an experimental study / 军事医学

Objective To verify the efficiency and stability of hydrogen-rich water preparation with hydrogen-rich rods. Methods ①Seven firenew hydrogen-rich rods were separately placed in seven plastic bottles, each filled with distilled water and soaked for 6 h, before the hydrogen concentration of the water was measured.This process was repeated 10 times.②After the hydrogen-rich rods with the strongest and weakest hydrogen product capacity were removed, the remaining 5 hydrogen-rich rods were placed separately into 5 plastic bottles filled with distilled water,put in a water bath pot at 20,40 and 60℃, respectively, and kept for 2, 4, 6, 8 and 10 h, respectively.Then, the hydrogen concentration, oxidation-reduction potential(ORP),and dissolved oxygen concentration(DO) were measured at various time points.③In order to determine the hydrogen emission rate from the hydrogen-rich water, the hydrogen-rich rods were constantly kept in some samples and the others were removed.All the sample bottle caps were kept open during the experimental process, and the hydrogen concentration was measured at such time points as 0, 10 and 30 min, 1, 2, 5, 12, 24, 30, 48 and 72 h, respectively.Results ①The hydrogen-rich rods used in this study could well meet the requirements.②When the environment temperature was kept constant, the hydrogen concentration of the water was increased with the soaking time of the hydrogen-rich rods, and the ORP of the water was reduced.However, the DO of the water was decreased with the rise of the environment temperature.③When the hydrogen-rich water was kept in opened plastic bottles with a 25 mm oral diameter, the hydrogen concentration of the samples with the hydrogen-rich rods reserved was almost about 0.50 ppm until 72 h, and that of the others was reduced to almost 0 ppm.Conclusion Our results demonstrate that the hydrogen-rich rods test is a simple and effective method for preparing hydrogen-rich water, which will be an valuable and useful method for using hydrogen-rich water in health promotion and prevention of chronic diseases.

Effect of Nutrients on Biological Nitrification.

Biological nitrification is the most commonly used process for nitrogen removal from wastewater. Nitrification is carried out in two steps. First ammonia is converted to nitrite by ammonia oxidizing bacteria. In the second step nitrite oxidizing bacteria convert nitrite to nitrate. The study involves the effect of nutrients (both organic and inorganic components) on biological nitrification and the optimum concentrations of di-sodium hydrogen phosphate, potassium di-hydrogen phosphate, sodium hydrogen phosphate, sucrose and ferric chloride were observed over ammonium ion removal. The effect of dissolved oxygen also was studied and maximum percentage removal of ammonium ion was found to be 89.2%.

Effects of the interannual variations in the flood pulse mediated by hypoxia tolerance: the case of the fish assemblages in the upper Parana River floodplain

The effects of the duration of the floods on abiotic variables and attributes of species tolerant and intolerant to hypoxia (STH and SIH respectively) were evaluated in rivers and lakes of the upper Paraná River. Fish were sampled once a year, in six sampling stations, during the high water period from 2000 to 2003. There were overall reductions in dissolved oxygen levels and increases in transparency of water in lakes in years of moderate floods. The duration of floods influenced species differentially based on their tolerance to hypoxia: in moderate floods, richness of STH increased and numerical abundance and biomass of SIH reduced significantly. Opposite relationships were detected between dissolved oxygen and the attributes of STH and SIH. Dissolved oxygen was the best predictor of variability of STH and SIH in years of moderate floods, whereas water transparency predicted significant amounts of STH in years of short floods. Being positively affected by dissolved oxygen reductions, STH seem to take advantages in persisting in seasonally harsh lentic habitats. The incorporation of abiotic data as well the differential tolerance of species to hypoxia would improve further investigations of the effects of interannual variations in the flood pulse on tropical fish assemblages.

Os efeitos da duração da cheia sobre algumas variáveis abióticas e atributos das assembleias de peixes (espécies tolerantes e intolerantes à hipóxia, STH e SIH, respectivamente) foram avaliados em rios e lagoas do alto rio Paraná. Os peixes foram amostrados uma vez ao ano, em seis localidades, durante o período de águas altas entre 2000 e 2003. Houve reduções generalizadas nos níveis de oxigênio dissolvido e aumentos na transparência da água em lagos em anos de cheias moderadas. A duração das cheias influenciou as espécies diferencialmente em função de sua tolerância à hipóxia: em anos de cheias moderadas, a riqueza das STH aumentou e a abundância numérica e de biomassa das SIH reduziram significativamente. Relações opostas foram detectadas entre o oxigênio dissolvido e os atributos das STH e SIH. O oxigênio dissolvido foi o melhor preditor da variabilidade de STH e SIH em anos de cheias moderadas, enquanto a transparência da água explicou uma significativa proporção das STH em anos de cheias curtas. Por serem positivamente afetadas pelas reduções no oxigênio dissolvido, as STH parecem ter vantagens em persistir em ambientes lênticos sazonalmente inóspitos. A incorporação de dados abióticos bem como da tolerância diferencial das espécies à hipóxia deve aprimorar as investigações dos efeitos das variações interanuais no pulso de inundação nas assembleias de peixes tropicais.

Long-term evolution of an activated sludge system under low dissolved oxygen conditions.

This study aims to evaluate the impacts of low dissolved oxygen (DO) levels on activated sludge performance. Two sequencing batch reactors (SBR), each with a working volume of 10 l, were investigated with different DO levels. Compared with high DO conditions (above 2 mg l-1 on average), low DO conditions in the SBR did not result in poor sludge bulking, lower chemical oxygen demand, and ammonium removal efficiency. Moreover, simultaneous nitrification and denitrification (SND) via nitrite and shortcut nitrification-denitrification were carried out under low DO levels. The average efficiencies of SND and nitrite accumulation ratios (NAR) in reactors A and B were 10.6 and 60.4%, respectively, under high DO levels and 4.1 and 76%, respectively, under low DO levels. Scanning electron microscopy revealed that the main types of bacteria in reactor A were spherical and short rod-shaped bacteria, whereas those in reactor B were long rod-shaped bacteria and filamentous bacteria. Thus, the appropriate DO concentration created excellent microbial community structures, which helped the biological systems to perform well under low DO level operating conditions.

Formation of aerobic granular sludge under adverse conditions: Low DO and high ammonia.

In this study, two adverse environments: low dissolved oxygen (DO) and high ammonia concentration, were employed to investigate the morphology, interspecies quorum sensing, extracellular polymers (EPS) characterization and microbial communities in the formation of aerobic granular sludge. Results showed that low DO could promote filamentous bacterial outgrowth. Under high ammonia concentration aerobic granular sludge (AGS) could still be cultivated, although it was looser and lighter than the control group. During the early stage of the AGS cultivation process, AI-2 activity reached a peak value in all three reactors, and ultrasonic pre-treatment was not beneficial to the release of AI-2. During AGS formation, the production of polysaccharide exhibited increases from 12.2 % to 40.3 %, 49.6 %, and 29.3 %. And PS in R2 was the highest as the result of sludge bulking. PS/PN was 1.5~8 in the three reactors. Three-dimensional EEM fuorescence spectroscopy variation indicated the change of protein in EPS, and the highest intensity of Peak T1 was obtained. The location shift of Peak T1 was not obvious, and Peaks A, C, and T2 shifted toward longer wavelengths (red shift) of 5~60 nm, or shorter wavelengths (blue shift) of 10~25 nm on the emission scale and / or excitation scale in all three reactors. This provided spectral information on the chemical structure changes. Bacteria in R3 had the highest species diversity, and all bacteria in b-Proteobacteria were identified as genus Thauera, which suggested that simultaneous nitrification and denitrification occurred in R3. The filamentous bacteria in seed sludge and R2 were species-richer. There was a low abundance of filamentous bacteria in R1 and R3, which contributed to the granule structure stability.

Impact of upwelling events on the sea water carbonate chemistry and dissolved oxygen concentration in the Gulf of Papagayo (Culebra Bay), Costa Rica: Implications for coral reefs / Impacto de los eventos de surgencia en la química del carbonato del agua de mar y la concentración de oxígeno disuelto en el Golfo de Papagayo (Bahía Culebra), Costa Rica: Implicaciones para los arrecifes de coral

The Gulf of Papagayo, Pacific coast of Costa Rica, is one of the three seasonal upwelling areas of Mesoamerica. In April 2009, a 29-hour experiment was carried out at the pier of the Marina Papagayo, Culebra Bay. We determined sea surface temperature (SST), dissolved oxygen concentration, salinity, pH, and the partial pressure of CO2 (pCO2). The aragonite saturation state (Ωa) as well as the other parameters of the marine carbonate system such as the total dissolved inorganic carbon (DIC) and the total alkalinity (TA) were calculated based on the measured pH and the pCO2. The entrainment of subsurface waters raised the pCO2 up to 645 µatm. SSTs, dissolved oxygen concentrations decreased form 26.4 to 23.7°C and from 228 to 144 µmol l-1. Ωa dropped down to values of 2.1. Although these changes are assumed to reduce the coral growth, the main reef building coral species within the region (Pocillopora spp. and Pavona clavus) reveal growth rates exceeding those measured at other sites in the eastern tropical Pacific. This implies that the negative impact of upwelling on coral growth might be overcompensated by an enhanced energy supply caused by the high density of food and nutrients and more favorable condition for coral growth during the non-upwelling season.

El Golfo de Papagayo, costa Pacífica de Costa Rica, es una de las tres regiones de afloramiento estacional de Mesoamérica. Las características físicas y químicas del agua que aflora no habían sido estudiadas. Durante 29 horas en Abril 2009, se estudiaron la temperatura superficial del mar (TSM), la concentración de oxígeno disuelto, salinidad, pH y la presión parcial de CO2 (pCO2), en la Marina Papagayo, Bahía Culebra. Con base en las mediciones de pH y pCO2 se calculó el estado de saturación de la aragonita (Ω) y otros parámetros del sistema de carbonatos como lo es el carbono orgánico disuelto (COD) y la alcalinidad total (AT). Los resultados indican que el arrastre por convección del agua sub-superficial durante los eventos de afloramiento aumenta la pCO2 y disminuye la TSM, la concentración de oxígeno disuelto y Ω. Aunque se asume que estas condiciones reducen el crecimiento coralino, las principales especies constructoras de arrecife en la región de Papagayo (Pocillopora spp. y Pavona clavus) tienen las mayores tasas de crecimiento en el Pacífico Tropical Oriental. Esto posiblemente implica que el efecto negativo del afloramiento es compensado por el crecimiento durante la época de no afloramiento.