Resveratrol-Loaded Attalea funifera Oil Organogel Nanoparticles: A Potential Nanocarrier against A375 Human Melanoma Cells.

This study aimed to evaluate Attalea funifera seed oil with or without resveratrol entrapped in organogel nanoparticles in vitro against A375 human melanoma tumor cells. Organogel nanoparticles with seed oil (SON) or with resveratrol entrapped in the seed oil (RSON) formed functional organogel nanoparticles that showed a particle size <100 nm, polydispersity index <0.3, negative zeta potential, and maintenance of electrical conductivity. The resveratrol entrapment efficiency in RSON was 99 ± 1%. The seed oil and SON showed no cytotoxicity against human non-tumor cells or tumor cells. Resveratrol at 50 µg/mL was cytotoxic for non-tumor cells, and was cytotoxic for tumor cells at 25 µg/mL. Resveratrol entrapped in RSON showed a decrease in cytotoxicity against non-tumor cells and cytotoxic against tumor cells at 50 µg/mL. Thus, SON is a potential new platform for the delivery of resveratrol with selective cytotoxic activity in the treatment of melanoma.

Well-dispersed titanium dioxide and silver nanoparticles on external and internal surfaces of asymmetric alumina hollow fibers for enhanced chromium (VI) photoreductions.

Heterogenous photocatalysis is a suitable alternative for wastewater treatment. The supporting of the solid catalyst in a porous material is suggested to facilitate catalyst recovery and reuse. Here we propose for the first time the evaluation of supporting silver (Ag)-decorated titanium dioxide (TiO2) catalysts on internal and external surfaces of alumina hollow fibers with asymmetric pore size distribution. The produced catalysts were considered for Cr(VI) photoreductions. The ultrasound-assisted process potentialized the distribution of Ag nanoparticles on the TiO2 surface. The loading of Ag nanoparticles at concentrations greater than 5 wt% was necessary to improve the TiO2 activity for Cr(VI) photoreduction. The loading of Ag nanoparticles at 30 wt% improved the Cr(VI) photoreduction of the single TiO2 catalyst from 40.49 ± 0.98 to 55.00 ± 0.83% after 180 min of reaction. Suspended and supported Ag-decorated TiO2 catalysts achieved total Cr(VI) photoreduction after 21 h of reaction. The adjusted reaction rate constant with the externally supported Ag-TiO2 catalyst was 3.57 × 10-3 ± 0.18 × 10-3 min-1. Similar reaction rate constants were achieved with suspended and internally supported catalysts (approximately 2.70 × 10-3 min-1). After 10 sequential reuses, all catalysts presented similar Cr(VI) photoreductions of approximately 66%. Nevertheless, the use of the externally supported catalyst is suggested for Cr(VI) photoreductions due to its superior catalyst activity at least in the first reuse cycles.

Nutritional influences on biomass behaviour and metabolic products by Chlamydomonas reinhardtii.

The recent works have shown the unicellular green alga Chlamydomonas reinhardtii is a relevant model for investigations of algal bioprocesses. In the current work, several media were evaluated in batch mode for a better understanding of C. reinhardtii metabolism. Nutrient-suppression using heterotrophic and mixotrophic conditions were performed. The findings showed C. reinhardtii metabolized lactose (from milk whey permeate) resulting in high biomass density (2.08 g/L) and total chlorophyll content (86.74 mg/m3). It was observed a specific growth rate of 0.023 h and 29 h for the doubling time. In sulfur-suppression, the algal growth (1.17 g/L) was reduced even though a carbon source (glucose) has been supplemented. Also, the specific growth rate (0.022 h) and the doubling time (31 h) was verified. The production of ethanol was slight and the acetic acid-suppression affected the C. reinhardtii performance providing slow cell growth (0.004 h) and high doubling time (154 h).

Antimicrobial activity of Annona muricata leaf oleoresin.

Bioactive compounds extracted from plants such as antimicrobials have attracted the attention of consumers and the food industry. This study aimed to determine the antimicrobial activity and chemical composition of Annona muricata leaf oleoresin obtained by supercritical CO2 extraction. The oleoresin was obtained by supercritical CO2 extraction and the chemical identification by gas chromatography coupled to mass spectrometry. Antimicrobial activity was evaluated by broth microdilution method against 14 foodborne fungi and bacteria. The oleoresin major chemical class was phytosterols (22.7%) and the major compounds were γ-sitosterol (15.7%), α-tocopherol (13.7%), phytol (13.1%), and hexadecanoic acid (11.5%). Minimum inhibitory concentration against bacteria ranged from 0.0025 to 0.010 mg mL-1. The oleoresin had high bactericidal activity against all bacteria, mainly Enterobacter cloacae and Pseudomonas aeruginosa with 0.005 mg mL-1 minimum bactericidal concentration. However, it had low fungicidal activity. The leaf oleoresin of A. muricata has promising applications in food, cosmetic, and pharmaceutical industries.

Chromatography-Independent Fractionation and Newly Identified Molecular Features of the Adzuki Bean (Vigna angularis Willd.) ß-vignin Protein.

Adzuki seed ß-vignin, a vicilin-like globulin, has proven to exert various health-promoting biological activities, notably in cardiovascular health. A simple scalable enrichment procedure of this protein for further nutritional and functional studies is crucial. In this study, a simplified chromatography-independent protein fractionation procedure has been optimized and described. The electrophoretic analysis showed a high degree of homogeneity of ß-vignin isolate. Furthermore, the molecular features of the purified protein were investigated. The adzuki bean ß-vignin was found to have a native size of 146 kDa, and the molecular weight determined was consistent with a trimeric structure. These were identified in two main polypeptide chains (masses of 56-54 kDa) that are glycosylated polypeptides with metal binding capacity, and one minor polypeptide chain with a mass 37 kDa, wherein these features are absent. The in vitro analysis showed a high degree of digestibility of the protein (92%) and potential anti-inflammatory capacity. The results lay the basis not only for further investigation of the health-promoting properties of the adzuki bean ß-vignin protein, but also for a possible application as nutraceutical molecule.

Photoreduction of chromium(VI) in microstructured ceramic hollow fibers impregnated with titanium dioxide and coated with green algae Chlorella vulgaris.

Here we propose an innovative photocatalytic hybrid system for the reduction of hexavalent chromium (Cr(VI)) from aqueous solutions. The hybrid system was composed of titanium dioxide (TiO2) immobilized in the micro-voids of asymmetric alumina hollow fibers and of the green algae Chlorella vulgaris coated on the outer sponge-like layer of the fiber. The photoreduction of Cr(VI) was systematically studied in different systems: single systems with TiO2 or algae; the synergistic system of algae combined with TiO2; and the proposed hybrid system composed of TiO2 and algae supported in ceramic hollow fibers. Morphological and energy dispersive spectroscopy analyses showed that TiO2 and the algae were properly supported in the substrate (alumina hollow fibers). For an initial Cr(VI) concentration of 10 mg L-1 and dosages of 1 g L-1 of TiO2 and algae, the hybrid system resulted in total Cr(VI) reduction after 16 h of process. Additionally, the efficiency of the hybrid system for Cr(VI) reduction was reduced in only 9% after 5 cycles of reuse and in 42% after 10 cycles of reuse. Thus, micro-structured ceramic hollow fibers impregnated with TiO2 and decorated with the green algae C. vulgaris was efficient for Cr(VI) reductions.

Dark fermentation effluent as substrate for hydrogen production from Rhodobacter capsulatus highlighting the performance of different fermentation systems.

Hydrogen production by biological route is a potentially sustainable alternative. Nowadays, energy production from sustainable sources has become urgent for several countries as well as for international policies. In this perspective, hydrogen has gained substantial global attention as clean, sustainable, and versatile energy carrier. In the current work, the resulting effluent from dark fermentation, rich in organic acids, was used as substrate for the purple non-sulfur bacteria (PNS) Rhodobacter capsulatus. In the first stage, experiments were carried out in bioreactors of 50 mL to check the influence of the composition of the effluent dark fermentation. The results proved that the provision of a sugar source improved bio-H2 production. The lactose and lactic acid concentrations exceeding 4.4 and 12 g/L, respectively, resulted in a productivity of up to 37.14 mmol H2/L days. Based on initial conditions obtained on the previous assays, in the second stage, a photo-fermentation in enlarged scale (1.5 L) was performed with the purpose to monitor the production of hydrogen and metabolites, sugar consumption and growth cells during the process. It was observed that the maximum productivity obtained was 98.23 mmol H2/L days in 26 h of process.

Alternative techniques to improve hydrogen production by dark fermentation.

The production of biofuels as an alternative to the fossil fuels has been mandatory for a cleaner and sustainable process. Hydrogen is seen as the fuel of the future because it has a very high energy density and its use produces only water instead of greenhouse gases and other exhaust pollutants. The biological synthesis of hydrogen by dark fermentation complies with these criteria. In the current work, the use of cheese whey permeate was evaluated aiming hydrogen production by dark fermentation using a microbial consortium in the semi-continuous process, with a reaction volume of 700 mL. The volume of the medium renewal and the frequency of replacements of fresh medium were evaluated to extend the production of H2. It is important to note decreases in the hydrogen production after 84 h. The target-product content became higher particularly when 466 mL of medium were withdrawn, in every 24 h in the first two replacements and, subsequently, in every 12 h. Besides, it was observed lower lactic acid concentration under this condition, suggesting that the shorter removal time of the medium could inhibit lactic acid bacteria, which may secrete bacteriocins that inhibit the hydrogen-producing microorganisms.

Evaluation of the temperature effect on vegetable oils by chemical analysis and ultraviolet­visible spectroscopy / Avaliação do efeito da temperature em óleos vegetais por analyses químicas e espectroscopia ultravioleta visível

Several methodologies could be used to characterize vegetable oil besides estimating thermal modification provided by high temperatures. These techniques are used as a proper tool to determine compositional and functional analysis of food ingredients and finished products. In general, vegetable oils are extracted from seeds like the rapeseed (canola oil) soybean (soybean oil), among others. In the present study vegetable oils such as canola and soybean were heated at a temperature of 100°C from 1h up to 28 h and the degraded products were measured to assess the oil stability at temperature. The determination of acid number, peroxide value and iodine number by chemical analysis was carried out for the estimation of the oxidative heat stability of these oils. Ultraviolet­visible spectroscopy strategy was also used to better comprehend this phenomenon, since it greatly improve the performance of measurements, in order to step up sensitivity. The findings demonstrated that vegetable oils thermal deteriorated as seen in the batocromic displacement of the samples heated and increasing the specific absorption in 350 nm. These data were capable to highlight the differences observed in the degree of unsaturation of oils.

Inúmeras metodologias podem ser utilizadas para caracterizar óleos vegetais, além de estimar modificações térmicas ocasionadas por altas temperaturas. Estas técnicas são usadas como ferramentas específicas para determinar a composição e função de ingredientes em alimentos e em produtos finais processados. Em geral, óleos vegetais são extraídos de sementes como a colza ou soja, entre outros. No presente trabalho, óleos vegetais como canola e soja foram aquecidos a temperatura de 100 °C entre 1 e 28 horas e os produtos de degradação foram medidos para avaliar a estabilidade do óleo na temperatura. As determinações da acidez, índice de peróxido e índice de iodo por análises químicas foram realizadas para estimar a estabilidade e a oxidação térmica dos óleos. A espectroscopia ultravioleta visível foi uma estratégia também utilizada para melhor compreender este fenômeno, uma vez que esta técnica aumenta o desempenho nas quantificações e quando se objetiva o incremento na sensibilidade. Os resultados demonstraram a deterioração térmica dos óleos vegetais através do deslocamento batocrômico das amostras aquecidas e o aumento da absorção a 350 nm. Estes dados ressaltam as diferenças observadas no grau de insaturação dos óleos.

In vitro and in silico studies of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase inhibitory activity of the cowpea Gln-Asp-Phe peptide.

Previous studies have shown that cowpea protein positively interferes with cholesterol metabolism. In this study, we evaluated the ability of the fraction containing peptides of <3 kDa, as well as that of the Gln-Asp-Phe (QDF) peptide, derived from cowpea ß-vignin protein, to inhibit HMG-CoA reductase activity. We established isolation and chromatography procedures to effectively obtain the protein with a purity above 95%. In silico predictions were performed to identify peptide sequences capable of interacting with HMG-CoA reductase. In vitro experiments showed that the fraction containing peptides of <3 kDa displayed inhibition of HMG-CoA reductase activity. The tripeptide QDF inhibits HMG-CoA reductase (IC50 = 12.8 µM) in a dose-dependent manner. Furthermore, in silico studies revealed the binding profile of the QDF peptide and hinted at the molecular interactions that are responsible for its activity. Therefore, this study shows, for the first time, a peptide from cowpea ß-vignin protein that inhibits HMG-CoA reductase and the chemical modifications that should be investigated to evaluate its binding profile.

Conventional and volatile salts as precipitating agents for recombinant antiviral protein / Sais convencionais e voláteis como agentes de precipitação para proteína recombinante com ação antiviral

Recombinant proteins expressed in cell culture have been shown to be relevant in the biopharmaceutical production focusing human health. The current work investigated the precipitation process of recAVLOEc protein, synthesized by E. coli BL21 (DE3) pLysS cells. The system is used for the AVLO expression that shown antiviral activity and it was found in the hemolymph of Lonomia obliqua caterpillar. The precipitation was conducted by the use of conventional salts (ammonium sulfate and sodium sulfate) and the volatile ammonium carbamate salt. Initially, the precipitated protein obtained from bacterial lysate was added to L929 cells to evaluate the cytotoxic effect; and besides Vero cells were infected with measles virus to verify the antiviral action of the precipitated recombinant protein. Toxic effect on the culture of L929 cells was observed for the precipitate obtained by the use of ammonium sulfate and sodium sulfate. In addition, tests in L929 cell cultures infected with EMC virus showed that samples of precipitated protein by salts did not show antiviral action. In Vero cell cultures, the precipitated protein by sodium sulfate showed antiviral action for measles virus.

Proteínas recombinantes expressas em culturas celulares têm se mostrado importantes na produção de fármacos de interesse para a saúde humana. Este estudo investigou a precipitação da proteína recAVLOEc, sintetizada por células de E. coli BL21 (DE3) pLysS, utilizadas como sistema de expressão da AVLO, proteína com atividade antiviral, originalmente encontrada na hemolinfa da lagarta Lonomia obliqua. A precipitação foi conduzida por meio do uso de sais convencionais (sulfato de amônio e de sódio) e do sal volátil carbamato de amônio. Inicialmente o precipitado proteico obtido do lisado bacteriano foi administrado em culturas de células L929 para avaliar o efeito citotóxico e posteriormente em células Vero infectadas com o vírus do sarampo, para a verificação da ação antiviral. Um efeito tóxico em culturas de L929 foi observado para os precipitados obtidos pelo uso de sulfato de amônio e de sódio. Testes em culturas de L929 infectadas com o vírus EMC foram também efetuados e as amostras de proteínas precipitadas com os sais convencionais e o sal volátil não resultaram em ação antiviral. Em culturas de células Vero, o uso do sulfato de sódio como agente de precipitação das proteínas contidas no lisado bacteriano resultou em ação antiviral para o sarampo.

Conventional and volatile salts as precipitating agents for recombinant antiviral protein

Recombinant proteins expressed in cell culture have been shown to be relevant in the biopharmaceutical production focusing human health. The current work investigated the precipitation process of recAVLOEc protein, synthesized by E. coli BL21 (DE3) pLysS cells. The system is used for the AVLO expression that shown antiviral activity and it was found in the hemolymph of Lonomia obliqua caterpillar. The precipitation was conducted by the use of conventional salts (ammonium sulfate and sodium sulfate) and the volatile ammonium carbamate salt. Initially, the precipitated protein obtained from bacterial lysate was added to L929 cells to evaluate the cytotoxic effect; and besides Vero cells were infected with measles virus to verify the antiviral action of the precipitated recombinant protein. Toxic effect on the culture of L929 cells was observed for the precipitate obtained by the use of ammonium sulfate and sodium sulfate. In addition, tests in L929 cell cultures infected with EMC virus showed that samples of precipitated protein by salts did not show antiviral action. In Vero cell cultures, the precipitated protein by sodium sulfate showed antiviral action for measles virus.

Replacement of sugars to hydrogen production by Rhodobacter capsulatus using dark fermentation effluent as substrate.

Hydrogen is a promising alternative for the increased global energy demand since it has high energy density and is a clean fuel. The aim of this work was to evaluate the photo-fermentation by Rhodobacter capsulatus, using the dark fermentation effluent as substrate. Different systems were tested by changing the type of sugar in the dark fermentation, investigating the influence of supplementing DFE with sugar and adding alternate and periodically lactose and glucose throughout the process. The supplementation of the DFE with sugar resulted in higher H2 productivity and the replacement of the sugars repeatedly during the photo-fermentation process was important to maintain the cell culture active. By controlling the residual amount of sugar, bacteria inhibition was avoided; lactic acid, that was toxic to the biomass, was consumed and the metabolic route of butyric acid production was predominant. Under optimum conditions, the H2 productivity reached 208.40mmolH2/Ld in 52h.

Prospective technology on bioethanol production from photofermentation.

The most important global demand is the energy supply from alternative source. Ethanol may be considered an environmental friendly fuel that has been produced by feedstock. The production of ethanol by microalgae represent a process with reduced environmental impact with efficient CO2 fixation and requiring less arable land. This work studied the production of ethanol from green alga Chlamydomonas reinhardtii through the cellular metabolism in a light/dark cycle at 25 °C in a TAP medium with sulfur depletion. The parameters evaluated were inoculum concentration and the medium supplementation with mixotrophic carbon sources. The combination of C.reinhardtii and Rhodobacter capsulatus through a hybrid or co-culture systems was also investigated as well. C.reinhardtii maintained in TAP-S produced 19.25±4.16 g/L (ethanol). In addition, in a hybrid system, with medium initially supplemented with milk whey permeated and the algal effluent used by R. capsulatus, the ethanol production achieved 19.94±2.67 g/L.

Behavior of wild-type and transfected S2 cells cultured in two different media.

An animal protein-free medium composed of IPL-41 containing 6 g L(-1) yeastolate ultrafiltrate, 10 g L(-1) glucose, 2 g L(-1) lactose, 5 g L(-1) glutamine, 1% lipid emulsion, and 0.1% Pluronic F-68 was used for producing recombinant proteins in batch mode employing two cell lines, S2AcRVGP2k expressing the G glycoprotein from rabies virus (RVGP) and S2AcHBsAgHy-9C expressing the surface antigen of hepatitis B virus (HBsAg), both obtained from Drosophila melanogaster S2 cells. Growth of wild-type S2 cells was also evaluated in the same medium. Cell behavior in the tested medium was compared to that verified in Sf900 II®. The results show that in shake flasks, S2AcRVGP2k and S2AcHBsAgHy-9C cells reached around 2 × 10(7) cells mL(-1) in both media. In supplemented IPL-41 and Sf900 II® media, S2AcRVGP2k cells produced 367 ng RVGP mL(-1) and 638 ng RVGP mL(-1), respectively, while S2AcHBsAgHy-9C cells correspondently produced 573 ng HBsAg mL(-1) and 322 ng HBsAg mL(-1) in the mentioned media. In stirred tanks, S2AcRVGP2k cells reached 3 × 10(7) cells mL(-1) and produced up to 758 ng RVGP mL(-1). In general, glucose was consumed by cells, while lactate and ammonia were produced.

Behavior of wild-type and transfected S2 cells cultured in two different media

An animal protein-free medium composed of IPL-41 containing 6 g L−1 yeastolate ultrafiltrate, 10 g L−1 glucose, 2 g L−1 lactose, 5 g L−1 glutamine, 1% lipid emulsion, and 0.1% Pluronic F-68 was used for producing recombinant proteins in batch mode employing two cell lines, S2AcRVGP2k expressing the G glycoprotein from rabies virus (RVGP) and S2AcHBsAgHy-9C expressing the surface antigen of hepatitis B virus (HBsAg), both obtained from Drosophila melanogaster S2 cells. Growth of wild-type S2 cells was also evaluated in the same medium. Cell behavior in the tested medium was compared to that verified in Sf900 II®. The results show that in shake flasks, S2AcRVGP2k and S2AcHBsAgHy-9C cells reached around 2 × 107 cells mL−1 in both media. In supplemented IPL-41 and Sf900 II® media, S2AcRVGP2k cells produced 367 ng RVGP mL−1 and 638 ng RVGP mL−1, respectively, while S2AcHBsAgHy-9C cells correspondently produced 573 ng HBsAg mL−1 and 322 ng HBsAg mL−1 in the mentioned media. In stirred tanks, S2AcRVGP2k cells reached 3 × 107 cells mL−1 and produced up to 758 ng RVGP mL−1. In general, glucose was consumed by cells, while lactate and ammonia were produced.

Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium.

The recombinant G glycoprotein from the surface of the rabies virus (RVGP) is a promising candidate as a rabies vaccine component and also for diagnostic purposes. In this study, RVGP production by transfected Drosophila melanogaster S2 cells cultivated in a serum-free medium (supplemented IPL-41 medium) was carried out. The effects of pH and pO(2) were evaluated in batch culture in parallel spinner flasks. The use of a pH equal to 6.3 and a pO(2) of 40% air saturation resulted in the highest RVGP content. These conditions were also used in fed-batch mode, yielding a RVGP content level of 98g/10(7) cells. The main nutrients consumed were glucose, glutamine, asparagine, serine and proline and the major metabolites produced were alanine and ammonia, according to the metabolism studies performed. Since RVGP is a transmembrane protein, two different methods for protein recovery were assessed and compared. Detergent-based cell disruption showed to be more effective than mechanical disruption with glass beads for glycoprotein recovery.

Formulation of a protein-free medium based on IPL-41 for the sustained growth of Drosophila melanogaster S2 cells.

An animal protein-free medium was developed for Drosophila melanogaster S2 (S2AcGPV2) cells genetically modified to produce the rabies virus G glycoprotein (GPV). IPL-41, used as a basal medium, was supplemented with yeastolate, carbohydrates, amino acids and lipids aiming initially to reduce and further to eliminate the need of fetal bovine serum. The S2AcGPV2 cells were fully capable of growing in serum-free supplemented IPL-41 medium containing 6 g L(-1) yeastolate ultrafiltrate, 10 g L(-1) glucose, 3.5 g L(-1) glutamine, 0.5 g L(-1) fructose, 2 g L(-1) lactose, 0.6 g L(-1) tyrosine, 1.48 g L(-1) methionine and 1% (v/v) lipid emulsion, reaching 19 x 10(6) cells mL(-1). Maximum specific growth rate and cell productivity were 0.025 h(-1) and 0.57 x 10(5) cells mL(-1) h(-1), respectively. Glucose and lactose were consumed during cell culture, but not fructose. Lactate concentration generally decreased during cell culture, while ammonium concentration reached 167 mg L(-1), however, without noticeable deleterious effects on cell growth. GPV concentration values achieved were, however, modest in the proposed medium formulation.

Evaluation of concentrated milk whey as a supplement for SF9 Spodoptera frugiperda cells in culture

Insect cell culture has become increasingly useful for the production of heterologous proteins as well as of baculovirus polyhedra, and several different culture media formulations can be employed for this purpose. The goal of this work was to assess the potential of lyophylized milk whey ultra filtration retentate (MWR) when associated to yeast extract (YE), glucose and Pluronic F68 (PF68) to partially replace fetal calf serum (FBS) in Spodoptera frugiperda Sf9 cells culture in Grace's medium, aiming AgMNPV baculovirus production. Batch cultivation results showed that the yeast extract and the milk whey concentrate effectively increased cell concentration to about half the level commonly verified for the Sf900II serum-free medium. A 4-fold increase in viable cell concentration was achieved when employing 5 percent (w/v) MWR, 8 g/L YE, 1 percent (v/v) of FBS and 2.7 g.l-1 of glucose, resulting in 1.6 x 10(7) polyhedra.ml-1 after infection with baculovirus.

Enhancement of Sf9 Cells and Baculovirus Production Employing Grace's Medium Supplemented with Milk Whey Ultrafiltrate.

Animal cells can be cultured both in basal media supplemented with fetal bovine serum (FBS) and in serum-free media. In this work, the supplementation of Grace's medium with a set of nutrients to reduce FBS requirements in Spodoptera frugiperda (Sf9) cell culture was evaluated, aiming the production of Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) at a cost lower than those for the production using Sf900 II medium. In Grace's medium supplemented with glucose, Pluronic F68 (PF68) and yeast extract (YE), the effects of FBS and milk whey ultrafiltrate (MWU) on cell concentration and viability during midexponential and stationary growth phase were evaluated. In spite of the fact that FBS presented higher statistical effects than MWU on all dependent variables in the first cell passage studies, after cell adaptation, AgMNPV polyhedra production was comparable to that in Sf900 II. Batch cultivation in Grace's medium with 2.7 g l(-1) glucose, 8 g l(-1) YE and 0.1% (w/v) PF68 supplemented with 1% (w/v) MWU and 3% (v/v) FBS increased viable cell concentration to about 5-fold (4.7x10(6) cells ml(-1)) when compared to Grace's containing 10% (v/v) FBS (9.5x10(5) cells ml(-1)). AgMNPV polyhedra (PIBs) production was around 3-fold higher in the MWU supplemented medium (1.6x10(7) PIBs ml(-1)) than in Grace's medium with 10% FBS (0.6x10(7) PIBs ml(-1)). This study therefore shows a promising achievement to significantly reduce FBS concentration in Sf9 insect cell media, keeping high productivity in terms of cell concentration and final virus production at a cost almost 50% lower than that observed for Sf900 II medium.