Biomass production and essential oil yield in Ageratum conyzoides L. - Asteraceae in different stages of development

An experiment was carried out at Sâo Manuel Experimental Farm, FCA-UNESP, Botucatu-SP, in 1994 and 1995, aiming to determine the biomass production and essential oil yield of mentrasto (Ageratum conyzoides L. - Asteraceae) in different stages of development. The statistical design used was a complete randomized block with 8 replications and 3 treatments (pre-flowering, flowering and post-flowering). The spacing was 70 cm between rows and 50 cm between plants, with 70 plants/parcel. The seedlings were transplanted 40 days after sowing. After harvesting, the fragments (roots, stems, leaves, flowers) and whole plants were washed, separated and weighted. The extractions of essential oil were done by hydrodistillation, using Clevenger apparatus, for two hours. There were statistic differences (Tukey 5%) in fresh weight of all vegetal parts w'hen harvested in different development stages. About essential oil yield, statistic differences were not verified among the treatments.

Influence of organic and chemical fertilization on the yield of flowers, contents and composition of essential oil of (Chamomilla recutita (L.) Rauschert)

A field experiment was conducted with chamomile (Chamomilla recutita [L.] Rauschert), in an area of the Olericulture and Medicinal Plants of the Horticulture Department at UNESP - Jaboticabal Campus, with the aim to evaluate the influence of organic and chemical fertilization on the yield of flowers, and content and composition of the essential oil of chamomile. The experimental design for the yield of flowers consisted of randomized blocks with 7 treatments and 4 replications, for the analysis of the contents and composition of the oil, the completely randomized block was used and for analysis of the correlation between harvesting and treatment, the split-plot design into randomized blocks was used. The treatments tested were: no fertilization, green manure (Mucuna aterrima + Crotalaria spectabilis), green manure (plant cocktail), organic fertilizer (farmyard manure), N as urea, N as ammonium sulphate, NPK with N supplement as ammonium sulphate. There was no influence of the treatments on the yield of flowers nor on the essential oil content; on the other hand both characteristics did show significant differences in harvesting times (Tukey 5%). The main yield was 885.90 kg/ha dry flowers and the mean oil content was 0,86%. The green manure treatment (M. aterrima + C. spectabilis) showed a higher percentage of chamazulene content, with a highly significant difference in harvesting times (Tukey 1%). The a-bisabolol percentages did not evidence significant differences between treatments. However, among harvesting times, there was a variation. A negative correlation was verified between the chamazulene and abisabolol percentages; the first increasing - from 21.02 to 36.17% - and the latter decreasing - from 14.12 to 8.72 % - from the first to the sixth harvest. The observed mean content of chamazulene was 14.64 % and a-bisabolol was 16.72 %.

Drying temperature effects in peppermint essential oil content and composition

Studies were conducted to show the effect of different temperatures in the drying process on the amount and quality of essential oils of peppermint (Mentha piperita L.) The leaves were harvested in the Demeter Farmer, Botucatu, SP, Brazil in december, 1997. The leaves were dried at 40°C, 60°C and 80°C, until establishment of the weights. The essential oil was extracted by destilation in Clevenger apparatus and analysed by GC-MS. Higher drying temperature sharply decreased the essential oil content (% v/w) from 1.0% (40°C) to 0.14% (60°C) and 0.12% (80°C). Higher drying temperatures also affected the composition, decreasing the contents of 1,8 cineol and citronelal until 80°C, and increasing the contents of menthol and neomenthol until 60°C.

Content of essential oil of fennel seeds in two different stages of development

This experiment was carried out at Plant Production Sector, Agronomical Science College-Botucatu, S.P., Brazil, in March, 2000. The aim of this assay was to determine the yield of essential oil of fennel (Foeniculum vulgare Miller) in different stages of development. Essential oils were prepared by hydrodistillation from the seeds using of Clevenger apparatus. The water utilized for the extraction of essential oil was sufficient to cover 100 g of seeds and the mixture was distilled for three hours. The volume of essential oil in the graduated side -arm of Clevenger apparatus was observed. There were no significative difference statistic was observed (Tukey 5%) in percentage (v/m) of oil content, based on dry weight of green seeds compared with dry weight of mature seeds, when they were harvested in two different stages of development. There was significative difference statistic between data obtained of humidity content of green seeds when these were compared with mature seeds. These results shows that others specifics studies about adaptation of fennel in tropical conditions are necessary, because the obtained data were different of data described on literature.

Influence of organic fertilisation on leaves and essential oil production of Ocimum gratissimum L.

The experiment was installed in Lageado Experimental Farm with aim to verify the influence of organic fertilisation (0, 4, 8 and 12 kg of manure/m2 - with four replications) on leaves and essential oil production of Ocimum gratissimum. The harvesting was done twice (May - autumn and August - winter), and the leaves were separated for extraction of essential oil by Clevenger apparatus. The leaf production and oil content were calculated on dry mass basis. The results showed no statistical difference for organic fertilisation, although significant difference was verified for seasons. The main constituents of essential oil were eugenol and 1,8-cineole. The amount of the eugenol was higher in autumn, while the presence of other components including 1,8-cineole, β-selinene and trans-caryophyllene were more dominant at wintertime.

Blue-greenish acrochaetioid algae in freshwater habitats are Chantransia stages of Batrachospermales sensu lato (Rhodophyta)

Fourteen culture isolates of freshwater acrochaetioid algae from distinct regions around the world were analysed, including the reddish species Audouinella hermannii, the dubious blue-greenish species A. pygmaea, and Chantransia stages from distinct taxonomic origins in the Batrachospermales sensu lato (Batrachospermaceae, Lemaneaceae and Thoreaceae). Four isolates (two 'Chantransia' stages and two species of Audouinella, A. hermannii and A. pygmaea) were tested under experimental conditions of temperature (10-25°C), irradiance (65 and 300 μmol photons m-2 s-1) and photoperiod (16:8 h and 8:16 h light/dark cycles). Plant colour is proposed as the only vegetative character that can be unequivocally applied to distinguish Audouinella from 'Chantransia', blue-greenish representing Chantransia stages and reddish applying to true Audouinella species (also forming reproductive structures other than monosporangia, e.g. tetrasporangia). Some isolates of A. pygmaea were proven to be unequivocally 'Chantransia stages owing either to production of juvenile gametophytes or to derivation from carpospores. No association of the morphology of A. pygmaea was found with any particular species, thus it should be regarded as a complex involving many species of the Batrachospermales sensu lato, as is also the case with A. macrospora. We therefore recommend that all blue-greenish acrochaetioid algae in freshwater habitats be considered as Chantransia stages of members of the Batrachospermales, and that the informal descriptors pygmaea and macrospora be used to distinguish the two discernable morphologies. Induction of gametophytes occurred under much wider conditions than previously reported, reinforcing the conclusion that requirements are probably species-specific. Although phenotypic plasticity was in evidence, with temperature, irradiance and photoperiod affecting morphology, no alga showed variation outside the limits based on traditional taxonomic studies. No overall trend was observed for vegetative or reproductive characters in response to temperature, irradiance and photoperiod for all the algae tested, only for specific algae or characters. Effects of temperature and irradiance on morphological characters were more evident, as well as strong interactions between these variables, whereas few differences were generally found in response to photoperiod and irradiance.

Antimicrobial activity of the essential oils of some spice herbs

Essential oils were obtained from fennel seeds, dill, cumin and coriander. Their antimicrobial activity was tested on isolated clinical specimens of patients treated at the University Hospital of the School of Medicine of Botucatu, SP, UNESP. Microorganisms were grown in BHI (Brain Heart Infusion/Oxoid) at 37oC/18 hours and resuspended in 0,5 Mac Farland's Standard (1,5 x 108 CFU/mL). The diffusion method was performed, putting 10 μl of the essential oils on paper disks (6mm of diameter) (Blank Disks/CECON) at 37oC/24 hours. After this period, the disks were put on plates containing Mueller Hinton Agar (Oxoid) and inoculated with the microorganisms. After 48 hours at 37oC, inhibitory zones were measured (mm) for the respective oils and strains. The essential oil from Anethum graveolens showed antimicrobial activity against Staphylococcus aureus (inhibitory zone=18 mm), Salmonella sp. (=11 mm) and E. coli (10 mm). The Cuminum cyminum essential oil was effective against E. coli, P. aeruginosa and Salmonella sp. and their inhibitory zones were 18, 10 and 23 mm, respectively. Coriandrum sativum oil was active only against Salmonella sp. (18 mm) and Foeniculum vulgare inhibited only E. coli (9 mm).

Essential oil composition of Aloysia triphylla (L'Herit) Britton leaves cultivated in Botucatu, São Paulo, Brazil

The family Verbenaceae comprises about 175 genera and 2300 species, distributed in tropics and subtropics, mainly in temperate zone of southern hemisphere. The lemon verbena (Aloysia triphylla (L'Herit) Britton) is a perennial, bushy plant originally from South America. The essential oil of this plant is used in pharmaceutical, cosmetic and perfumery industry. Therapeutic properties include febrifuge, sedative, stomachical, diuretic, and antispasmodic activities. The present work aimed to identify the chemical composition of essential oil of Aloysia triphylla leaves. The study was done in Lageado Experimental Farm of the Department of Plant Production-Horticulture, Agronomical Sciences College, São Paulo State University Campus of Botucatu. Leaves of lemon verbena from Medicinal and Aromatic Plant Garden, were collected in the end of winter (September/2001). The essential oil was extracted by hydrodistillation, in Clevenger apparatus. 100 g of leaves were used in each extraction. Four extractions were performed during three hours. The essential oils of the leaves were analyzed in Gas Chromatography Mass spectrometry (CG-MS, Shimadzu, QP-5000), equipped with capillary column DB-5 (30 m × 0,25 mm × 0,25 mm), split 1/35, injector for 220 C°, detector for 230 C°, dragged by gas He (1,0 mL/min), with programmed temperature for 60 C° to 240 C°, 3 C°/min. The identification of the substances was held by comparison of their mass spectra with data of the CG-MS (Nist 62 lib), literature references and retention index of Kovats. The main constituents of essential oils were geranial (29.54 %), neral (27.01 %), limonene (15.93 %), geranyl acetate (4.0 %) and geraniol (3.96 %). This species possesses high quantity of monoterpenes and low quantity of sesquiterpenes.

Composition of coriander essential oil from Brazil

Coriander (Coriandrum sativum L.) is an annual and herbaceous plant, belonging to the Apiaceae family. Native of southern Europe and western Mediterranean region, this herb is cultivated world widely. This species, rich in linalool, has potential using as source of essential oil and as a medicinal plant. It has been used as analgesic, carminative, digestive, depurative, anti-rheumatic and antispasmodic agent. Its fruits (commonly called seeds) are used for flavoring candies, in cookery, perfumery, beverage and in tobacco industry. The aim of this study was to analyze the chemical composition of the seed essential oil of this species grown in Botucatu, São Paulo, Brazil. The experiment was carried out in Lageado Experimental Farm, Department of Plant Production, Agronomical Sciences College, São Paulo State University. The fruits were harvest 108 days after sowing. The essential oils were extracted by hydro distillation, in Clevenger apparatus. 50 g of fruits were used in each extraction. Three extractions were performed during three hours. The essential oils were analyzed in Gas Chromatography Mass Spectrometer (CG-MS, Shimadzu, QP-5000), equipped with DB-5 capillary column (30 m × 0,25 mm × 0,25 mm), split 1/20, injector for 240 C°, detector for 230 C°, dragged by gas He (1,7 mL/min), with programmed temperature for 40 C° (5 min)-150 C°, 4 C°/min; 150 C°-280 C°, 8 C°/min. The identification of the compounds was made by comparison of their spectra of masses with data from CG-MS (Nist 62 lib), literature references and retention index of Kovats. The 18 most important components were identified and quantified. The main components of the oil were linalool (77.48 %), γ-terpinene (4.64 %), α-pinene (3.97 %), limonene (1.28 %), geraniol (0.64 %) and 2-decenal (0.16 %).

Electrophysiological responses of Atta sexdens rubropilosa workers to essential oils of Eucalyptus and its chemical composition

The leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 is the most harmful of the Eucalyptus pests, causing severe losses in wood production through defoliation. Various strategies have been tried and effort spent on the development of methods to control this pest, however no practical and environmentally acceptable one currently exists. In this work the chemical composition of the essential oil of seven Eucalyptus species was identified and the selectivity and sensitivity of antennal receptors of A. sexdens rubropilosa workers to the volatile compounds were determined using the electroantennographic technique (EAG and GC-EAD). Analysis by GC-EAD showed in E. cloesiana and E. maculata, respectively, seventeen and sixteen terpenes that elicited responses in ant workers' antennae, indicating the potential role of the essential oils as allelochemicals that determine the choice of the foraging material. © 2006 Verlag der Zeitschrift für Naturforschung.

Nerolidol, an antiulcer constituent from the essential oil of Baccharis dracunculifolia DC (Asteraceae)

In this study, the antiulcerogenic effect of essential oil from Baccharis dracunculifolia was evaluated using the model of acute gastric lesions induced by ethanol. The ulcerative lesion index (ULI) was significantly reduced by oral administration of the essential oil of B. dracunculifolia at doses of 50, 250 and 500 mg/kg which reduced the lesions by 42.79, 45.70 and 61.61%, respectively. The analysis of the chemical composition of the essential oil from B. dracunculifolia by GC showed that this was composed mainly of mono- and sesquiterpenes and the majority compound was nerolidol. Therefore, antiulcerogenic activity of nerolidol (50, 250 and 500 mg/kg) was investigated using ethanol-, indomethacin- and stress-induced ulcer models in rat. In the stress-induced ulcer model, a significant reduction of the ULI in animals treated with nerolidol (50, 250 and 500 mg/kg) and cimetidine (100 mg/kg) was observed, compared to the control group (p < 0.05). The percentage of inhibition of ulcer was 41.22, 51.31, 56.57 and 53.50% in groups treated with 50, 250, 500 mg/kg of nerolidol and 100 mg/kg of cimetidine (positive control), respectively. Regarding ethanol- and indomethacin-induced ulcer models, it was observed that the treatment with nerolidol (250 and 500 mg/kg) significantly reduced the ULI in comparison with the control group (p < 0.05). The dose of 50 mg/kg reduced the parameters analyzed but this was not statistically significant. In the ethanol-induced model percentage of inhibition of ulcer was 34.20, 52.63, 87.63 and 50.87% in groups treated with 50, 250, 500 mg/kg of nerolidol and 30 mg/kg of omeprazol (positive control), respectively. In indomethacin-ulcer the percentage of inhibition of ulcer was 34.69, 40.80, 51.02 and 46.93% in groups treated with 50, 250, 500 mg/kg of nerolidol and 100 mg/kg of cimetidine (positive control), respectively. The results of this study show that nerolidol displays antiulcer activity, as it significantly inhibited the formation of ulcers induced in different animal models. However, further pharmacological and toxicological investigations, to delineate the mechanism(s) of action and the toxic effects, are required to allow the use of nerolidol for the treatment of gastric ulcer. © 2007 Verlag der Zeitschrift für Naturforschung.

Exploratory study on sequestration of some essential metals by indigo carmine food dye

Indigo carmine forms a stable complex with different ions, and the stability constant of the complexes were evaluated as log K equal to 5.75; 5.00; 4.89 and 3.89 for complexes with Cu(II), Ni(II), Co(II) and Zn(II) ions, respectively, in 0.1 mol L -1 carbonate buffer solution at pH 10. The interaction between Cu(II) ions and indigo carmine (IC) in alkaline medium resulted in the formation of the Cu 2(IC) complex, measured by the spectrophotometric method, with a stoichiometric ratio between indigo carmine and metal ions of 2:1 (metal-ligand). The reported method has also been successfully tested for determination of copper in pharmaceutical compounds based on copper-gluconate without pre-treatment.

Anatomical features of the urethra and urinary bladder catheterization in female mice and rats. An essential translational tool

PURPOSE: To present fundamental anatomical aspects and technical skills necessary to urethra and urinary bladder catheterization in female mice and rats. METHODS: Urethral and bladder catheterization has been widely utilized for carcinogenesis and cancer research and still remains very useful in several applications: from toxicological purposes as well as inflammatory and infectious conditions to functional aspects as bladder dynamics and vesicoureteral reflux, among many others. RESULTS: Animal models are in the center of translational research and those involving rodents are the most important nowadays due to several advantages including human reproducibility, easy handling and low cost. CONCLUSIONS: Although technical and anatomical pearls for rodent urethral and bladder access are presented as tackles to the advancement of lower urinary tract preclinical investigation in a broaden sight, restriction to female animals hampers the male microenvironment, demanding future advances.

Yield and phytochemical characterization of essential oil from Ocimum selloi B. obtained by hydrodistillation and supercritical fluid extraction

The yield and chemical composition of essential oils from leaves of Ocimum selloi B. submitted to organic and mineral fertilization, obtained by hydrodistillation and supercritical fluid extraction (SFE) were compared. Essential oil was extracted in a Clevenger-type apparatus for 2 h 30 min and analyzed by GC-MS (Shimadzu, QP 5050-DB-5 capillary column - 30 m × 0.25 mm × 0.25 μm). Carrier gas was helium (1.7 ml/min); split ratio: 1:30. Temperature program: 50°C, rising to 180°C at 5°C/min, 180°C, rising to 280°C at 10°C/min. Injector temperature: 240°C and detector temperature: 230°C. Identifications of chemical compounds were made by matching their mass spectra and Kovat's indices (IK) values with known compounds reported in the literature. An Applied Separations-apparatus (Speed SFE, model 7071, Allentown, PA, EUA) was used for SFE extractions. They were conducted at pressure 200 bar and temperature 30°C (20 min in static mode and 40 min in dynamic mode). The supercritical CO2 flow rate was (6.8±0.7)×10-5 kg-CO2/s. The essential oil collected was immersed in ethylene glycol bath (5°C). The yield of essential oils obtained by SFE was larger than hydrodistillation in both fertilization treatments (279 and 333% for organic and mineral fertilizations, respectively). There were no differences between the fertilization treatments. The amount of the volatile components showed by GC-MS chromatogram was highest in the essential oil obtained by hydrodistillation than SFE. The main volatile constituents of the essential oils were trans-anethole (Hydrodistillation: organic - 52.4%; mineral - 55.0%/ SFE: Hydrodistillation - 62.8%; mineral - 66.8%) and methyl-chavicol (Hydrodistillation: organic - 37.3%; mineral - 38.3%/ SFE: organic - 8.4%; mineral - 4.3%). A reduction of methyl-chavicol relative proportion of essential oil obtained by SFE was observed. Cys-anethole, α-copaene, trans-cariofilene, germacrene-D, β-selinene, biciclogermacrene and spathulenol were expressed only in hydrodistillation. The extraction of essential oil by SFE presented larger yield of essential oil than hydrodistillation technique, presenting, however, these essential oils, different phytochemical profiles.

Biomass production and chemical composition of essential oil of Piper callosum as affected by spacing in Manaus, Amazonas State, Brazil

The family Piperaceae contains nearly 2000 species, most of them distributed in two genera, Piper and Peperomia. In Brazil circa 170 Piper species are found, mainly in tropical areas Their use ranges from flavoring and culinary to raw material for pharmaceutical and cosmetic industry. One of these species, Piper callosum, is used in folk medicine in the Amazon area. The objective of this study was to evaluate the production of biomass (aerial parts) as well as yield and composition of the essential oil from the leaves, according to different spacing between cultivated plants at Embrapa Western Amazon, in Manaus, State of Amazonas, Brazil. The experimental design was randomized blocks, with four treatments and seven replicates with six plants. Biomass production was inversely proportional to the spatial arrangements, with the greatest biomass production (1034.93 kg/ha) in the shortest spacing (E1), although no statistical difference was verified between E3 and E4. The same response was observed for the production of essential oil. The chemical composition of the oil was not affected by spacing, and major compounds found were safrole (59.1%), beta-pinene (8.3%), alpha-pinene (6.5%), methyl eugenol (6.3%) and 1,8-cineole (4.1).