Fatty acid ethyl esters production using a non-commercial lipase in pressurized propane medium

The objective of this work is to investigate the production of fatty acid ethyl esters from soybean oil in compressed propane using a non-commercial lipase from Yarrowia lipolytica and two commercial ones as catalysts, Amano PS and Amano AY30. The experiments were performed in the temperature range of 35-65 °C. at 50 bar, enzyme concentration of 5 wt%, oil to ethanol molar ratio of 1:6 and 1:9, and solvent to substrates mass ratio of 2:1 and 4:1. The results indicated that low reaction conversions were generally obtained with the use of commercial and non-commercial lipases in pressurized propane medium. On the other hand, the aspects of low solvent to substrates mass ratio and mild temperature and pressure operating conditions used to produce ethyl esters justify further investigations to improve reaction yields.

Docosahexaenoic acid ethyl esther (DHAEE) microcapsule production by spray-drying: optimization by experimental design

Docosahexaenoic acid is an essential polyunsaturated fatty acid with important metabolic activities. Its conjugated double bonds make it susceptible to decomposition. Its stability may be improved through fatty acid entrapment with a spray-drying technique; however, the many parameters involved in this technique must be considered to avoid affecting the final product quality. Therefore, this study aimed to evaluate the entrapment conditions and yields of fish oil enriched with docosahexaenoic acid ethyl ester. Microcapsules were obtained from Acacia gum using a spray-drying technique. The experimental samples were analyzed by chromatography and delineated by Statistica software, which found the following optimum entrapment conditions: an inlet temperature of 188 °C; 30% core material; an N2 flow rate of 55 mm; and a pump flow rate of 12.5 mL/minute. These conditions provided a 66% yield of docosahexaenoic acid ethyl ester in the oil, corresponding to 19.8% of entrapped docosahexaenoic acid ethyl ester (w/w). This result was considered significant since 30% corresponded to wall material.

Crescimento de dois cultivares de soja submetidos a aplicações de herbicidas e fungicidas

Existem relatos de que a aplicação de fungicidas do grupo das estrobilurinas na cultura da soja tem trazido aumento de produtividade mesmo em locais sem incidência de doenças, indicando que as alterações de ordem fisiológica devem ser quantificadas. Este estudo foi desenvolvido com o objetivo de avaliar o crescimento de dois cultivares de soja (MG/BR46 Conquista e BRS Valiosa RR) submetidos a aplicações de herbicidas e fungicidas (estrobilurinas e/ou triazóis). Dois experimentos foram desenvolvidos em campo com esquema de tratamentos em parcelas subdivididas no tempo (5 x 12), em que nas parcelas foram alocadas cinco combinações de herbicidas (glyphosate a 720 g ha-1 ou sethoxydim (230 g ha-1) + bentazon + chlorimuron-ethyl (480 + 11,25 g ha-1)) e fungicidas (pyraclostrobin a 75 g ha-1, epoxiconazole a 50 g ha-1 ou pyraclostrobin + epoxiconazole (66,5 + 25 g ha-1)); e como subparcelas foram consideradas as datas de avaliação entre 30 e 120 dias após semeadura. A aplicação de herbicidas não alterou o crescimento dos cultivares de soja. A aplicação de fungicidas do grupo das estrobilurinas associado a triazóis promoveu maior acúmulo de massa de matéria seca e área foliar. Ainda a aplicação desses fungicidas em R2 (pleno florescimento) e R5.1 (enchimento de grãos) resultou em maiores valores de taxas de crescimentos absoluto e relativo, assimilatória líquida e de crescimento da cultura em período próximo ao final do ciclo da cultura, diferenciando, inclusive, dos tratamentos com aplicação somente de triazóis.

Misturas em tanque com glyphosate para o controle de trapoeraba, erva-de-touro e capim-carrapicho em soja RR®

O uso de misturas de glyphosate, em tanque, para manejo de espécies de plantas daninhas de difícil controle tem sido prática comum entre os agricultores brasileiros. Desta forma, este trabalho teve como objetivo avaliar a eficácia e seletividade de misturas, em tanque, de herbicidas com glyphosate para o controle de trapoeraba (Commelina benghalensis L.), erva-de-touro (Tridax procumbens L.) e capim-carrapicho (Cenchrus echinatus L.) na cultura da soja RR®. O experimento foi conduzido em Maracaí, São Paulo, no período de novembro de 2006 a março de 2007, utilizando-se o cultivar CD-214RR® e delineamento experimental de blocos ao acaso, com 21 tratamentos e quatro repetições. Os tratamentos foram constituídos da aplicação de: glyphosate (180; 360; 540 e 720 g ha-1); glyphosate em sequencial (180/360; 360/360 e 540/360 g ha-1); glyphosate + chlorimuron-ethyl 360+10; 540+10; 360+5/ 360+5 g ha-1); glyphosate + lactofen (360+120; 540+120; 360+60/ 360+60 g ha-1); glyphosate + cloransulam-methyl (360+30; 540+30; 360+16,9/ 360+12,9 g ha-1); glyphosate + carfentrazone (360+4 g ha-1); glyphosate + imazethapyr (360+50 g ha-1); glyphosate + imazethapyr (177,8+30 g ha-1) e testemunhas capinada e sem capina. Apesar da similaridade de produtividade de grãos entre os tratamentos com glyphosate isolado e sequencial, nas doses 540, 720 e 540/ 360 g ha-1, as misturas em tanque com chlorimuron-ethyl, cloransulam-methyl, lactofen e imazethapyr favoreceram o controle de espécies de plantas daninhas tolerantes ao glyphosate como C. benghalensis e T. procumbens.

Herbicidas alternativos para o controle de Conyza sumatrensis (Retz.) E. H. Walker resistentes aos inibidores da ALS e EPSPs

RESUMO A buva (Conyza sumatrensis), uma das principais plantas daninhas já identificadas no sul do Brasil, vem apresentando controle insatisfatório, em resposta à aplicação dos herbicidas chlorimuron-ethyl e glyphosate. Por esta razão, o objetivo deste estudo foi avaliar herbicidas alternativos, visando ao controle de biótipos de C. sumatrensis, com resistência de nível baixo ao herbicida chlorimuron-ethyl e resistentes ao herbicida glyphosate. O experimento foi realizado em casa de vegetação, entre abril e agosto de 2012, no município de Passo Fundo, RS. O delineamento experimental foi o inteiramente casualizado, com quatro repetições, sendo avaliados 15 tratamentos com herbicidas, além de uma testemunha sem aplicação. As variáveis consideradas foram controle percentual, aos 14, 21 e 28 dias após a aplicação dos tratamentos (DAT) e a matéria seca da parte aérea, aos 28 DAT. Como resultados, observou-se que os biótipos foram 100% controlados, aos 28 DAT, pelos tratamentos alternativos de paraquat + diuron; ammonium glufosinate; glyphosate + 2,4-D; glyphosate + ammonium glufosinate; 2,4-D; tembotrione e tembotrione + atrazine. O biótipo 17 evidenciou menor sensibilidade aos herbicidas inibidores da ALS e os biótipos 05, 17 e 20 não foram controlados pelo herbicida glyphosate.

Effects of growth reducer and nitrogen fertilization on morphological variables, SPAD index, interception of radiation and productivity of wheat

ABSTRACT The objective of this study was to evaluate the effect of growth reducer and nitrogen fertilization on morphological variables, SPAD index, radiation interception, and grain yield of three cultivars of wheat. The experimental design was a randomized block in factorial scheme 3x5x2, with three cultivars (Mestre, Iguaçú and Itaipú), five nitrogen doses (0, 40, 80, 120, 160 Kg ha-1), and application or no application of a growth reducer, with three replications. The following characteristics were evaluated: plant height, SPAD index, leaf area index (LAI), Global Radiation Interception (GRI) and grain yield. The Tukey test (p < 0.05) was used for the comparison between the means of cultivar and growth reducer factors, and for a regression analysis to evaluate N levels. Increasing the dose of nitrogen promotes an increase in LAI of plants of wheat crops differently among cultivars, which leads to a greater degree of global radiation interception. At doses higher or equal to 120 Kg ha-1 of nitrogen, there are significant differences in grain yield between treatments with and without the application of the growth reducer. The significant interaction between growth reducer and nitrogen dose, showed that applications of growth reducer increase the GRI at doses above and below 80 Kg ha-1 of nitrogen. Nitrogen rates of 138 and 109 Kg ha-1 are responsible for maximum grain yields of wheat, which is 4235 and 3787 Kg ha-1 with and without the use of growth reducer, respectively.

In vitro action of some disinfectants on Paracoccidioides brasiliensis yeast forms

The fungicidal action of sodium hypochlorite (0.3, 1, 2.5, 5 and 10%); formaldehyde (2, 5, and 10%); and ethyl alcohol (70%) on yeast forms of Paracoccidioides brasiliensis Pb 18 and a newly-isolated Goiana strain was described. Contact between the fungus and the disinfectants was maintained for 1, 2, 24, 48 and 72 hours at room temperature. Viability was evaluated by the fluorescein diacetate-ethidium bromide treatment, culture in solid and liquid media (36ºC and 26ºC); yeast to mycelial germination at room temperature; and radiometric study of metabolic activity. All concentrations of disinfectants were found to be effective in inactivating Pb 18 and Goiana strains, except for the 1-hour contact with 2% formaldehyde, in which fluorescein diacetate-ethidium bromide treatment was found to reveal 40 and 27% of viable cells, respectively. The yeast to mycelial germination method was considered to reveal faster and similar results as compared to culture in solid and liquid media.

Contribution to the laboratory diagnosis of human cryptosporidiosis

Human cryptosporidiosis is an infection caused by Cryptosporidium spp, a coccidial protozoan of emergencial pathogenicity and responsible for severe and prostrating watery diarrhea, mainly in immunocompromised patients. Smears of stools submitted to concentration and staining by carbol fuchsin technique has been used in our laboratory as a diagnostic procedure for cryptosporidiosis. The long time observing the smears in the microscope due to the small size of the forms and the low contrast of the staining led us to introduce some modifications in the original protocol for the acid-fast staining. The smears were treated with the carbol fuchsin solution for 3 minutes as recommended by LENNETTE et al., 1985 and the solution of the ethyl alcohol 70%-chloride acid 0.5% was used instead of the ethyl alcohol-sulfuric acid 5% recommended by HENRIKSEN & POHLENZ, 1981. Smears were treated with the discoloration solution for 2 minutes. These modifications promoted a better washing out of the excess of carbol fuchsin therefore increasing the dye efficiency. In such conditions, the visualization of protozoan oocysts on the slides examined became easier. They were observed by the contrast of their intense pink-reddish pigmentation against the blue background. It's worthwhile to emphasize that these modifications offer advantages when time and accuracy are concerned.

Antibacterial activity of GUAVA, Psidium guajava Linnaeus, leaf extracts on diarrhea-causing enteric bacteria isolated from Seabob shrimp, Xiphopenaeus kroyeri (Heller)

Guava leaf tea of Psidium guajava Linnaeus is commonly used as a medicine against gastroenteritis and child diarrhea by those who cannot afford or do not have access to antibiotics. This study screened the antimicrobial effect of essential oils and methanol, hexane, ethyl acetate extracts from guava leaves. The extracts were tested against diarrhea-causing bacteria: Staphylococcus aureus, Salmonella spp. and Escherichia coli. Strains that were screened included isolates from seabob shrimp, Xiphopenaeus kroyeri (Heller) and laboratory-type strains. Of the bacteria tested, Staphylococcus aureus strains were most inhibited by the extracts. The methanol extract showed greatest bacterial inhibition. No statistically significant differences were observed between the tested extract concentrations and their effect. The essential oil extract showed inhibitory activity against S. aureus and Salmonella spp. The strains isolated from the shrimp showed some resistance to commercially available antibiotics. These data support the use of guava leaf-made medicines in diarrhea cases where access to commercial antibiotics is restricted. In conclusion, guava leaf extracts and essential oil are very active against S. aureus, thus making up important potential sources of new antimicrobial compounds.

Parasites in stool samples in the environment of Ilha da Marambaia, Rio de Janeiro, Brazil: an approach in public health

This research aimed to describe the frequency of parasites in stool samples in the environment of Ilha da Marambaia, Rio de Janeiro, Brazil. One hundred and five stool samples were collected and processed by the coproparasitological techniques ethyl acetate sedimentation and centrifuge-flotation using saturated sugar solution. Parasites were detected in 81.9% of the samples, hookworm being the most prevalent, followed by Trichuris vulpis. Ascaris sp. eggs were also found. A high level of evolutive forms of parasites with public health risk was found in stool samples of the environment studied. We propose that health education programs, allied to an improvement of human and animal health care, must be employed to reduce the environmental contamination.

APPLICATION OF 6-NITROCOUMARIN AS A SUBSTRATE FOR THE FLUORESCENT DETECTION OF NITROREDUCTASE ACTIVITY IN Sporothrix schenckii

Introduction Sporothrix schenckii is a thermal dimorphic pathogenic fungus causing a subcutaneous mycosis, sporotrichosis. Nitrocoumarin represents a fluorogenic substrate class where the microbial nitroreductase activity produces several derivatives, already used in several other enzyme assays. The objective of this study was the analysis of 6-nitrocoumarin (6-NC) as a substrate to study the nitroreductase activity in Sporothrix schenckii. Methods Thirty-five samples of S. schenckii were cultivated for seven, 14 and 21 days at 35 °C in a microculture containing 6-nitrocoumarin or 6-aminocoumarin (6-AC) dissolved in dimethyl sulfoxide or dimethyl sulfoxide as a negative control, for posterior examination under an epifluorescence microscope. The organic layer of the seven, 14 and 21-day cultures was analyzed by means of direct illumination with 365 nm UV light and by means of elution on G silica gel plate with hexane:ethyl acetate 1:4 unveiled with UV light. Results All of the strains showed the presence of 6-AC (yellow fluorescence) and 6-hydroxylaminocoumarin (blue fluorescence) in thin layer chromatography, which explains the green fluorescence observed in the fungus structure. Conclusion The nitroreductase activity is widely distributed in the S. schenckii complex and 6-NC is a fluorogenic substrate of easy access and applicability for the nitroreductase activity detection.

CHEMICAL CHARACTERIZATION AND EVALUATION OF ANTIBACTERIAL, ANTIFUNGAL, ANTIMYCOBACTERIAL, AND CYTOTOXIC ACTIVITIES OF Talinum paniculatum

SUMMARY In this study, the bioactivity of Talinum paniculatum was evaluated, a plant widely used in folk medicine. The extract from the T. paniculatum leaves (LE) was obtained by percolation with ethanol-water and then subjecting it to liquid-liquid partitions, yielding hexane (HX), ethyl acetate (EtOAc), butanol (BuOH), and aqueous (Aq) fractions. Screening for antimicrobial activity of the LE and its fractions was evaluated in vitro through broth microdilution method, against thirteen pathogenic and non-pathogenic microorganisms, and the antimycobacterial activity was performed through agar diffusion assay. The cytotoxic concentrations (CC90) for LE, HX, and EtOAc were obtained on BHK-21 cells by using MTT reduction assay. The LE showed activity against Serratia marcescens and Staphylococcus aureus, with Minimum Inhibitory Concentration (MIC) values of 250 and 500 µg/mL, respectively. Furthermore, HX demonstrated outstanding activity against Micrococcus luteus and Candida albicans with a MIC of 31.2 µg/mL in both cases. The MIC for EtOAc also was 31.2 µg/mL against Escherichia coli. Conversely, BuOH and Aq were inactive against all tested microorganisms and LE proved inactive against Mycobacterium tuberculosisand Mycobacterium bovisas well. Campesterol, stigmasterol, and sitosterol were the proposed structures as main compounds present in the EF and HX/EtOAc fractions, evidenced by mass spectrometry. Therefore, LE, HX, and EtOAc from T. paniculatumshowed potential as possible sources of antimicrobial compounds, mainly HX, for presenting low toxicity on BHK-21 cells with excellent Selectivity Index (SI = CC90/MIC) of 17.72 against C. albicans.

MOLLUSCICIDAL AND LARVICIDAL ACTIVITIES OF Atriplex inflata AERIAL PARTS AGAINST THE MOLLUSK Galba truncatula, INTERMEDIATE HOST OF Fasciola hepatica

Fasciolosis is a widespread parasitosis of farm live-stock in many developing countries. For this reason, it is necessary to search for new substances against parasitic diseases caused by flukes. Indeed, a wide variety of terrestrial plants have been subjected to chemical and pharmacological screening in order to discover their potential for human medicinal use. The molluscicidal and larvicidal activities of Atriplex inflata were tested on Galba truncatula and Fasciola hepatica larval stages infecting this snail in Tunisia. Phytochemical tests were conducted on extracts in order to establish a meaningful relationship with molluscicidal and larvicidal activities. The molluscicidal activity was evaluated by subjecting snails to sample aqueous solutions. Accordingly, hexane, ethyl acetate, methanol and methanol-water (8:2, v-v) were used as extraction solvents. As a result, hexane and ethyl acetate extracts showed potent activity, according to the World Health Organization, giving LC50 = 7.59 mg/L and 6.69 mg/L for hexane extracts of leaves and fruits, respectively. Ethyl acetate extracts gave LC50 = 5.90 mg/L and 7.32 mg/L for leaves and fruits, successively. Molluscicidal activities of powders were less potent on snails, but active according to the World Health Organization. Hexane and ethyl acetate extracts from leaves and fruits gave potent larvicidal activities with a delay rate exceeding 45.50% (45.50- 98.92%). Phytochemical tests showed that these activities may be attributed to the presence of triterpenoids and/or sterols.

Larvicidal effects of endophytic and basidiomycete fungus extracts on Aedes and Anopheles larvae (Diptera, Culicidae)

Introduction In vitro bioassays were performed to access the larvicidal activity of crude extracts from the endophytic fungus Pestalotiopsis virgulata (Melanconiales, Amphisphaeriaceae) and the saprophytic fungus Pycnoporus sanguineus (Basidiomycetes, Polyporaceae) against the mosquitoes Aedes aegypti and Anopheles nuneztovari. Methods The extracts were tested at concentrations of 100, 200, 300, 400 and 500ppm. Ethyl acetate mycelia (EAM) extracts and liquid culture media (LCM) from Pe. virgulata and Py. sanguineus were tested against third instar larvae of Ae. aegypti and An. nuneztovari. Results The larvicidal activity of the EAM extracts from Pe. virgulata against Ae. aegypti had an LC50=101.8ppm, and the extract from the basidiomycete fungus Py. sanguineus had an LC50=156.8ppm against the Ae. aegypti larvae. The Pe. virgulata extract had an LC50=16.3ppm against the An. nuneztovari larvae, and the Py. sanguineus extract had an LC50=87.2ppm against these larvae. Conclusions These results highlight the larvicidal effect of EAM extracts from the endophyte Pe. virgulata against the two larval mosquitoes tested. Thus, Pe. virgulata and Py. sanguineus have the potential for the production of bioactive substances against larvae of these two tropical disease vectors, with An. nuneztovari being more susceptible to these extracts.

Pimenta pseudocaryophyllus inhibits virulence factors and promotes metabolic changes in Candidayeast

IntroductionThis is the first study to examine the in vitrosusceptibility and the expression of virulence factors in Candida species in the presence of Pimenta pseudocaryophyllus (Gomes) L.R. Landrum (Myrtaceae), a Brazilian plant known as paucravo. Additionally, the mechanisms of action of the crude ethanol extract and the ethyl acetate and aqueous fractions of this plant were investigated.MethodsThe in vitro susceptibility of Candida was tested using the broth microdilution method, whereas an XTT reduction assay was used for biofilms. Adherence was determined by counting the number of yeast cells that adhered to 100 oral epithelial cells, and hyphal formation was verified in the hyphal induction medium M199. Flow cytometry with propidium iodide and FUN-1 was performed to assess the mechanism of action.ResultsThe results revealed that the crude ethanol extract and the ethyl acetate and aqueous fractions of P. pseudocaryophyllusinhibited the growth of Candida isolates at a minimal inhibitory concentration (MIC) ranging from 64 to 256µg/mL, whereas the 50% sessile minimal inhibitory concentration (SMIC50) ranged from 512 to >1,024µg/mL. Adherence and hyphal formation were significantly reduced in the presence of the crude ethanol extract and both fractions. Although cell membrane injury was detected, the predominant mechanism of action appeared to be the alteration of yeast metabolism, as demonstrated by flow cytometry.ConclusionsOur results indicated that antifungal activity reduced the expression of virulence factors in yeast via the alteration of yeast metabolism, suggesting that the crude extract of P. pseudocaryophyllus and its fractions may contain novel antifungal agents.