Fractionation and characterization of cell wall polysaccharides from coffee (Coffea arabica L.) pulp.

Cell wall polysaccharides were isolated by sequential extractions from coffee pulp, the main solid waste from coffee processing. Extractions were conducted with distilled water at room and boiling temperatures, 0.5 % ammonium oxalate and 0.05 M Na2CO3 to obtain pectic fractions. Hemicelluloses were extracted by using 2 M and 4 M NaOH. The composition of the hemicellulose fractions suggested the presence of xyloglucans, galactomannans and arabinogalactan-proteins (AGPs). The main part of the cell wall polysaccharides recovered from coffee pulp were pectins branched with arabinogalactans. Coffee pulp pectic fractions were low-methoxylated with various amounts of protein (0.5-8.4 %) and phenolics (0.7-8.5 %). Detection at 280 nm in the HPSEC analyses and radial gel diffusion assay using Yariv reagent indicated the presence of AGPs in most of these fractions. NMR analyses of chelating agent (CSP) and dialyzed water (WSPD) extracted pectins were carried out. The results demonstrated that CSP contains only AG I. On the other hand, AG I and AG II are present in WSPD, probably covalently linked to the pectic portion. Comparison with the literature indicated similarities between the cell wall polysaccharides from coffee pulp and green coffee beans.

Ocotea nutans (Nees) Mez: structural elucidation of C-hetorosides flavonoids and evaluation of their antioxidant and antibacterial properties from ethyl acetate extract.

The Ocotea genus exhibits a variety of pharmacological, antibacterial and antioxidant effects. The aim of the present study was to evaluate the antioxidant potential and antibacterial properties of the ethyl acetate fraction of Ocotea nutans leaves. Isolation and identification of the phenolic compounds from the fraction was also carried out. The isolated compounds were characterised by one- and two-dimensional nuclear magnetic resonance analysis and identified as vitexin (1) and isovitexin (2). The ethyl acetate fraction of Ocotea nutans leaves demonstrated considerable antioxidant potential. The observed minimum inhibitory concentration of 250 µg.mL-1 was classified as a moderate antibacterial activity against Enterococcus faecalis. Findings from this study demonstrate the utility of this plant as a potential source of antioxidant and antibacterial agents.

Preliminary evaluation of the larvicidal activity of extracts and fractions from Ocotea nutans (Nees) Mez against Aedes aegypti.

INTRODUCTION: Aedes aegypti is the main vector of dengue and yellow fever. Recently, the use of plant-sourced larvicides has gained momentum. METHODS: The hydroethanolic extracts and fractions ofOcotea nutansleaves and stems were bioassayed to determine the larvicidal efficacy of these samples. RESULTS: S-HEX (hexane fraction from the crude stem extract) demonstrated high potential for controlling third-stage larvae, with an LC50 of 14.14 µg.mL-1 (concentration required to inhibit 50% of the treated larvae). CONCLUSIONS: Extracts from O. nutans were effective against third-stage larvae ofA. aegyptiafter 24 h of exposure.

Preliminary evaluation of the larvicidal activity of extracts and fractions from Ocotea nutans (Nees) Mez against Aedes aegypti

Abstract INTRODUCTION: Aedes aegypti is the main vector of dengue and yellow fever. Recently, the use of plant-sourced larvicides has gained momentum. METHODS: The hydroethanolic extracts and fractions ofOcotea nutansleaves and stems were bioassayed to determine the larvicidal efficacy of these samples. RESULTS: S-HEX (hexane fraction from the crude stem extract) demonstrated high potential for controlling third-stage larvae, with an LC50 of 14.14 µg.mL-1 (concentration required to inhibit 50% of the treated larvae). CONCLUSIONS Extracts from O. nutans were effective against third-stage larvae ofA. aegyptiafter 24 h of exposure.

Chemical Characterization of Opuntia ficus-indica (L.) Mill. Hydroalcoholic Extract and Its Efficiency against Gastrointestinal Nematodes of Sheep.

Opuntia ficus-indica (L.) Mill. is a xerophylous plant that originated in tropical and subtropical America. This plant is popularly known in Brazil as "palma forrageira" (cactus pear) and plays a fundamental role in animal nutrition, mainly in the Northeastern semi-arid region of the country. The plant has several uses since it presents bioactive compounds that confer biological and pharmacological properties. In this context, the cactus pear can also be considered a potential product to combat parasite infections. The objective of this study was to chemically characterize the O. ficus-indica hydroalcoholic extract (OFIEOH) and to determine its efficacy against gastrointestinal parasites using in vitro tests. Initially, the hydroalcoholic extract from cladode peels of O. ficus-indica was produced by maceration for 21 days. For the chemical characterization, colorimetric dosages were performed for carbohydrates, proteins, phenols and condensed tannins. Liquid chromatography coupled to mass spectrometry/electron spray ionization (LC-MS/ESI) was used to characterize the polyphenolic profile of the OFIEOH extract. Fifteen compounds were identified in the OFIEOH extract, such as methyl, glycosylated and aglycone quercetin derivatives and aglycone and glycosylated kaempferol derivatives. Tri-glycosylated methyl quercetin derivatives were the main compounds identified. In vitro egg hatch (EHT) and larval migration tests (LMT) were used in a range of concentrations of OFIEOH from 12.5 to 100 mg/mL for EHT and 12.5 to 200 mg/mL for LMT. In addition, the LMT was used to test ivermectin (IVM) (from 11.4 to 57.1 µM), associated with the inhibitory concentration of 50% (IC50) for OFIEOH. The combination of OFIEOH (12.5 to 200 mg/mL) plus the IC50 of IVM was also tested. The efficacy of OFIEOH alone varied from 19.33 to 90.0% using the EHT. The LMT revealed an efficacy of 5.78 to 77.26% for the extract. Both tests showed a concentration-dependence inhibitory effect. We found a drug-extract antagonistic neutralizing effect when doses of IVM were added to OFIEOH (maximum efficacy of 73.78%), while a positive additive effect was observed when OFIEOH was added to the IC50 of IVM (IC50 of 82.79 for OFIEOH alone against an IC50 of 55.08 of OFIEOH + IVM). The data from this work indicate that OFIEOH alone may be considered as a suitable ecofriendly product to control gastrointestinal parasites of sheep, offering a more holistic approach to improve animal farming and welfare. The drug-extract interaction is also a promising therapeutic alternative, reducing the final dose to the host, with an optimum combination effect.

Assessment of anthelmintic activity and bio-guided chemical analysis of Persea americana seed extracts.

The aim of this study was to characterize the extracts and fractions of Persea americana Mill. (Avocado) seeds and to determine the composition and the in vitro anthelmintic activity against third-stage larvae (L3) of Haemonchus contortus. The fresh (F) and dried (H) avocado seeds (PA) were subjected to extraction with hot water (W-PAF, W-PAH), ethanol (E-PAF, E-PAH) or methanol 70% (v/v), and partition with solvents of increasing polarity [n-hexane (H-PAF, H-PAH), chloroform (C-PAF, C-PAH), ethyl acetate (Ea-PAF, Ea-PAH), and n-butanol (B-PAF, B-PAH)], yielding a total of 14 extracts/fractions. After considering the yield, water solubility, and the preliminary results of the larval migration test (LMT), the E-PAF, E-PAH, H-PAF, and H-PAH were selected for further experiments. E-PAH presented an efficiency concentration of 50% (EC50) of 36 µg/mL on the LMT. E-PAH showed the greatest efficiency when its EC50 was compared to the other fractions (E-PAF = 147 µg/mL; H-PAF = 801 µg/mL; H-PAH = 77 µg/mL). After that, the E-PAH was chemically characterized, considering its quantitative polyphenolic and flavonoid contents by colorimetric and chromatographic techniques. E-PAH presented 50, 38, and 24 mg/g of dry matter of total phenol, condensed tannins (CT), and flavonoid contents, respectively. Using high performance liquid chromatography (HPLC) analysis, E-PAH had shown to have epicatechin (4.7 µg/mL), rutin (2.8 µg/mL), and chlorogenic acid (1.4 µg/mL) as its main constituents besides quercetin. These isolated compounds were evaluated using the LMT in order to relate the composition to the anthelmintic activity observed for E-PAH. Quercetin (EC50 = 7.8 µg/mL) and epicatechin (EC50 = 10 µg/mL) presented a higher efficiency than rutin (EC50 = 30 µg/mL). Chlorogenic acid was also tested with the LMT but did not present a significant efficiency. According to the results, the phenolic composition of E-PAH and the EC50 values obtained for the isolated phenols, it can be suggested that, besides the CT content, the presence of epicatechin and rutin contributed to the larvicidal activity of E-PAH. In conclusion, avocado seeds may be used as a source of polyphenols with promising anthelmintic applications.

Efeito anticoccidiano de extrato hidroalcoólico de Artemisia annua em camas de aves contaminadas com Eimeria sp / Anticoccidial effect of Artemisia annua hydroalcoholic extract in poultry beds contaminated with Eimeria sp

O objetivo deste estudo foi avaliar a eficácia do extrato hidroalcoólico de Artemisia annua frente a oocistos de Eimeria sp. em camas contaminadas. O extrato foi produzido com 7 dias de percolação a 4°C, sendo posteriormente realizada a marcha fitoquímica; dosagem de fenóis totais, quantificação de artemisinina, ensaio antioxidante e teste de toxicidade. Para testar a atividade anticoccidiana, camas de aves compostas de cepilho de árvores foram contaminadas com 5000 oocistos. Foram formados quatro tratamentos, em triplicata, nos quais foram usadas diferentes concentrações, sendo G1: 12mg/mL, G2: 8mg/mL, G3: 4mg/mL e C-: água. Após a contaminação, foram aspergidos, 800 mL dos extratos nas diferentes concentrações sobre as camas e coletadas, em triplicatas, 10 cm2 de cada local, aleatoriamente, nos tempos: 0, 3, 6, 24, 48, e 72 horas após a aplicação. Nas análises fitoquímicas, foram evidenciados diversos compostos com propriedades antiparasitárias, como flavonoides e taninos. O fitoterápico continha 59,409±1,47μg/dL de artemisinina. O produto na concentração de 12mg.mL-1 apresentou eficácia entre 45,5 e 42,1%. Os resultados dos testes bioquímicos, juntamente com os encontrados no teste anticoccidiano, evidenciaram que o extrato produzido possui alto potencial para combater Eimeria sp.

The aim of this study was to determine the efficacy of hydroalcoholic extract of Artemisia annua against oocysts of Eimeria sp. in contaminated poultry beds. The extract was produced after 7 days of storage at 4°C, which was used to perform the phytochemical screening; the artemisinin measurement; the total phenolic; antioxidant testing and toxicity test. To test the anticoccidial activity, the birds space composed of shaver trees, were contaminated with 5000 oocysts. Four treatment were formed, in triplicate, were used in different concentrations as G1: 12mg/mL, G2:= 9mg/mL, G3: 6mg/mL, and C-: water. After contamination 800 mL of the herbal at different concentrations were sprayed on the bed and collected, in triplicate, 10 cm2 each site, randomly, at times: 0, 3, 6, 24, 48, and 72 hours after application. In phytochemical analysis, were shown compounds with antiparasitic properties, such as flavonoids and tannins. The herbal contained 59.409±1.47mg/dL artemisinin. The product at a concentration of 12mg.mL-1 showed efficacy from 44.25 to 40.71%. The results of biochemical tests, with the in vitro test showed that the extract has produced high potential for combating Eimeria sp.

Efeito anticoccidiano de extrato hidroalcoólico de Artemisia annua em camas de aves contaminadas com Eimeria sp / Anticoccidial effect of Artemisia annua hydroalcoholic extract in poultry beds contaminated with Eimeria sp

O objetivo deste estudo foi avaliar a eficácia do extrato hidroalcoólico de Artemisia annua frente a oocistos de Eimeria sp. em camas contaminadas. O extrato foi produzido com 7 dias de percolação a 4°C, sendo posteriormente realizada a marcha fitoquímica; dosagem de fenóis totais, quantificação de artemisinina, ensaio antioxidante e teste de toxicidade. Para testar a atividade anticoccidiana, camas de aves compostas de cepilho de árvores foram contaminadas com 5000 oocistos. Foram formados quatro tratamentos, em triplicata, nos quais foram usadas diferentes concentrações, sendo G1: 12mg/mL, G2: 8mg/mL, G3: 4mg/mL e C-: água. Após a contaminação, foram aspergidos, 800 mL dos extratos nas diferentes concentrações sobre as camas e coletadas, em triplicatas, 10 cm2 de cada local, aleatoriamente, nos tempos: 0, 3, 6, 24, 48, e 72 horas após a aplicação. Nas análises fitoquímicas, foram evidenciados diversos compostos com propriedades antiparasitárias, como flavonoides e taninos. O fitoterápico continha 59,409±1,47μg/dL de artemisinina. O produto na concentração de 12mg.mL-1 apresentou eficácia entre 45,5 e 42,1%. Os resultados dos testes bioquímicos, juntamente com os encontrados no teste anticoccidiano, evidenciaram que o extrato produzido possui alto potencial para combater Eimeria sp...

The aim of this study was to determine the efficacy of hydroalcoholic extract of Artemisia annua against oocysts of Eimeria sp. in contaminated poultry beds. The extract was produced after 7 days of storage at 4°C, which was used to perform the phytochemical screening; the artemisinin measurement; the total phenolic; antioxidant testing and toxicity test. To test the anticoccidial activity, the birds space composed of shaver trees, were contaminated with 5000 oocysts. Four treatment were formed, in triplicate, were used in different concentrations as G1: 12mg/mL, G2:= 9mg/mL, G3: 6mg/mL, and C-: water. After contamination 800 mL of the herbal at different concentrations were sprayed on the bed and collected, in triplicate, 10 cm2 each site, randomly, at times: 0, 3, 6, 24, 48, and 72 hours after application. In phytochemical analysis, were shown compounds with antiparasitic properties, such as flavonoids and tannins. The herbal contained 59.409±1.47mg/dL artemisinin. The product at a concentration of 12mg.mL-1 showed efficacy from 44.25 to 40.71%. The results of biochemical tests, with the in vitro test showed that the extract has produced high potential for combating Eimeria sp...

The involvement of PUMP from mitochondria of Araucaria angustifolia embryogenic cells in response to cold stress.

In this study, the responses of plant uncoupling mitochondrial protein (PUMP) and alternative oxidase (AOX) in mitochondria from embryogenic cells of A. angustifolia subjected to cold stress (4°C for 24 h or 48 h) is reported. In the mitochondria of stressed cells, PUMP activity increased by approximately 45% (at 24h and 48 h), which was determined by measuring the oxygen consumption after the addition of linoleic acid and the inhibition by BSA and ATP. PUMP activation was confirmed using transmembrane electrical potential (Δψ) assays. Immunoblot assays showed an increase of PUMP expression by 40% and 150% after 24h and 48 h of cold stress, respectively. AOX activity, measured under conditions similar to those of the PUMP assays, was only slightly increased in the mitochondria from stressed cells (at 24h and 48 h), as demonstrated by oxygen consumption experiments. Cell viability was unaffected by cold stress, indicating that the effects on PUMP and AOX were not caused by cell death. These results show that the main response of this gymnosperm to cold stress is the activation of PUMP, which suggests that this protein may be involved in the control of reactive oxygen species generation, which has been previously associated with this condition.

Arabinogalactan-proteins from cell suspension cultures of Araucaria angustifolia.

Arabinogalactan-proteins (AGPs), found in the culture medium of suspension cells of Araucaria angustifolia grown in plant growth regulator-free and plant growth regulator-containing BM media, BM0 and BM2, respectively, were evaluated quantitatively and qualitatively. The concentrated extracellular fractions (CEFs), obtained from suspension cell cultures grown for 20 days in BM0 and BM2 media yielded two fractions, CEF-0 and CEF-2, respectively. CEF-0 and CEF-2 was submitted to selective precipitation using the beta-glucosyl Yariv reagent (beta-GlcY) to isolate AGPs for structural characterization; this yielded fractions designated CEF-0YPF and CEF-2YPF, respectively. The monosaccharide composition analysis established that samples were composed of Rha, Ara, Gal and uronic acid in a molar ratio 3:37:55:5 (CEF-0YPF) and 1:37:58:4 (CEF-2YPF), although trace amounts (<0.5 mol%) of Xyl were also found. Methylation analysis of CEF-YPF fractions showed similar results for both CEF-0YPF and CEF-2YPF, with non-reducing terminal units of Araf, Arap, Galp, Rhap and Xylp, as well as 3-O-substituted and 5-O-substituted Araf units and 3-O-substituted, 6-O-substituted and 3,6-di-O-substituted Galp units. The amino acid composition analysis established Ser, Ala, and Hyp as major amino acids in both samples. In conclusion, this investigation has shown that CEF-0YPF and CEF-2YPF contain macromolecules having typical AGP characteristics, including a Hyp/Ala/Ser-rich protein moiety, a (1-->3) and/or (1-->6) linked beta-d-galactopyranosyl main chain substituted by Gal, Ara, Rha and Xyl residues, and binding affinity for beta-GlcY and monoclonal anti-AGP antibodies.