“Seaweed exudates and settlement behaviour of sedentary organisms"

The present investigation has looked exclusively into the aspect of the biological phenomenon of settling behaviour by two serious fouling offenders encountered in the tropical seas mainly on the hulls of ships and stationary structures in the harbours. The cue to study the behaviour was adopted from the observations so far made by scientists on the epizoic growth of these organisms on the surfaces of algal fronds of variegated shape, texture, size etc. The results do indicate that there are sufficient qualities of bioactive substances produced by plants occupying the lowest categories in organic evolution and curiously enough these substances have withstood the test of time.

Management approaches in organic potato and tomato production

The presented thesis considered three different system approach topics to ensure yield and plant health in organically grown potatoes and tomatoes. The first topic describes interactions between late blight (Phytophthora infestans) incidence and soil nitrogen supply on yield in organic potato farming focussing in detail on the yield loss relationship of late blight based on results of several field trials. The interactive effects of soil N-supply, climatic conditions and late blight on the yield were studied in the presence and absence of copper fungicides from 2002-2004 for the potato cultivar Nicola. Under conditions of central Germany the use of copper significantly reduced late blight in almost all cases (15-30 %). However, the reductions in disease through copper application did not result in statistically significant yield increases (+0 – +10 %). Subsequently, only 30 % of the variation in yield could be attributed to disease reductions. A multiple regression model (R²Max), however, including disease reduction, growth duration and temperature sum from planting until 60 % disease severity was reached and soil mineral N contents 10 days after emergence could explain 75 % of the observed variations in yield. The second topic describes the effect of some selected organic fertilisers and biostimulant products on nitrogen-mineralization and efficiency, yield and diseases in organic potato and tomato trials. The organic fertilisers Biofeed Basis (BFB, plant derived, AgroBioProducts, Wageningen, Netherlands) and BioIlsa 12,5 Export (physically hydrolysed leather shavings, hair and skin of animals; ILSA, Arizignano, Italy) and two biostimulant products BioFeed Quality (BFQ, multi-compound seaweed extract, AgroBioProducts) and AUSMA (aqueous pine and spruce needle extract, A/S BIOLAT, Latvia), were tested. Both fertilisers supplied considerable amounts of nitrogen during the main uptake phases of the crops and reached yields as high or higher as compared to the control with horn meal fertilisation. The N-efficiency of the tested fertilisers in potatoes ranged from 90 to 159 kg yield*kg-1 N – input. Most effective with tomatoes were the combined treatments of fertiliser BFB and the biostimulants AUSMA and BFQ. Both biostimulants significantly increased the share of healthy fruit and/or the number of fruits. BFQ significantly increased potato yields (+6 %) in one out of two years and reduced R. solani-infestation in the potatoes. This suggests that the biostimulants had effects on plant metabolism and resistance properties. However, no effects of biostimulants on potato late blight could be observed in the fields. The third topic focused on the effect of suppressive composts and seed tuber health on the saprophytic pathogen Rhizoctonia solani in organic potato systems. In the present study 5t ha-1 DM of a yard and bio-waste (60/40) compost produced in a 5 month composting process and a 15 month old 100 % yard waste compost were used to assess the effects on potato infection with R. solani when applying composts within the limits allowed. Across the differences in initial seed tuber infestation and 12 cultivars 5t DM ha-1 of high quality composts, applied in the seed tuber area, reduced the infestation of harvested potatoes with black scurf, tuber malformations and dry core tubers by 20 to 84 %, 20 to 49 % and 38 to 54 %, respectively, while marketable yields were increased by 5 to 25 % due to lower rates of wastes after sorting (marketable yield is gross yield minus malformed tubers, tubers with dry core, tubers with black scurf > 15% infested skin). The rate of initial black scurf infection of the seed tubers also affected tuber number, health and quality significantly. Compared to healthy seed tubers initial black scurf sclerotia infestation of 2-5 and >10 % of tuber surface led in untreated plots to a decrease in marketable yields by 14-19 and 44-66 %, a increase of black scurf severity by 8-40 and 34-86 % and also increased the amount of malformed and dry core tubers by 32-57 and 109-214 %.

Does fertilizer type and method of application cause significant differences in essential oil yield and composition in rosemary (Rosmarinus officinalis L.)?

Organic fertilizers based on seaweed extract potentially have beneficial effects on many crop plants. Herewe investigate the impact of organic fertilizer on Rosmarinus officinalis measured by both yield and oilquality. Plants grown in a temperature-controlled greenhouse with a natural photoperiod and a controlledirrigation system were treated with seaweed fertilizer and an inorganic fertilizer of matching mineralcomposition but with no organic content. Treatments were either by spraying on to the foliage or wateringdirect to the compost. The essential oil was extracted by hydro-distillation with a Clevenger apparatusand analysed by gas-chromatography mass-spectrometry (GC–MS) and NMR. The chemical composi-tions of the plants were compared, and qualitative differences were found between fertilizer treatmentsand application methods. Thus sprayed seaweed fertilizer showed a significantly higher percentage of�-pinene, �-phellandrene, �-terpinene (monoterpenes) and 3-methylenecycloheptene than other treat-ments. Italicene, �-bisabolol (sesquiterpenes), �-thujene, and E-isocitral (monoterpenes) occurred insignificantly higher percentages for plants watered with the seaweed extract. Each was significantly dif-ferent to the inorganic fertilizer and to controls. The seaweed treatments caused a significant increasein oil amount and leaf area as compared with both inorganic treatments and the control regardless ofapplication method.

Uso de Ascophyllum nodosum para o enraizamento de microestacas de eucalipto

Pós-graduação em Agronomia (Irrigação e Drenagem) - FCA

Aplicação de extratos de algas marinhas em cafeeiro sob deficiência hídrica e estresse salino

Pós-graduação em Agronomia (Irrigação e Drenagem) - FCA

Extratos de Ascophyllum nodosum no tratamento de sementes de milho e soja: avaliações fisiológicas e moleculares

A obtenção de uma lavoura com população adequada de plantas depende da utilização de diferentes práticas agronômicas, estando o sucesso condicionado ao uso de sementes de boa qualidade. No entanto, a semeadura dificilmente é realizada em condições ideais o que resulta em problemas na emergência das plantas. Grande é a procura por alternativas que melhorem a germinação e o desenvolvimento inicial dos cultivos resultando em uniformidade de emergência, garantindo o estande e culminando em produção. O tratamento de sementes com agroquímicos fitossanitários é uma solução parcial no combate de agentes fitopatológicos e pragas, mas poucas são as soluções adotadas para melhorar o desenvolvimento vegetal inicial, que pode levar a desuniformidade e falhas no estande gerando prejuízos econômicos. Os extratos de algas já demonstraram em diversos estudos sua eficiência no desenvolvimento vegetal quando aplicados em plantas. Porém poucos são os estudos voltados para os efeitos dos extratos de algas na germinação e emergência. Assim, o intuito deste trabalho foi testar o extrato comercial de Ascophyllum nodosum, e diferentes fracionamentos do mesmo, no tratamento de sementes de soja e milho. Avaliou-se o efeito de diferentes doses no desenvolvimento das plântulas e as doses de melhor resposta foram utilizadas no tratamento de sementes de soja a fim de associar as respostas obtidas à expressão gênica de 9 genes relacionados ao processo germinativo em 24 e 48h de embebição. Sementes de soja tratadas com o extrato comercial resultaram em plântulas menos desenvolvidas o que pode estar relacionado ao alto teor de sais contidos no produto. O tratamento com as demais frações favoreceu o desenvolvimento das plântulas, principalmente o desenvolvimento radicular. Sementes de milho tratadas não apresentaram desenvolvimento tão satisfatório quanto as sementes de soja tratadas. A análise da expressão gênica relativa demonstrou que o tratamento com frações do extrato comercial é capaz de regular algumas vias do metabolismo hormonal, como a isopentenil transferase e a GA20 oxidase 2, e do catabolismo de reservas, como a acil-CoA oxidase. Em condições ótimas, o tratamento de sementes de soja com frações do extrato comercial de A. nodosum favoreceu o desenvolvimento inicial das plântulas de soja, no entanto não ocasionou grandes alterações no desenvolvimento de milho. Este estudo demonstrou a possibilidade de utilização de frações do extrato de A. nodosum no favorecimento do desenvolvimento inicial de plântulas de soja. Maiores estudos são necessários quanto às respostas em campo e na atenuação de estresses para viabilizar seu uso como um bioestimulante em sementes.

Cellular and in-vitro models to assess antioxidant activities of seaweed extracts and the potential use of the extracts as ingredients

Seaweeds contain a range of antioxidant compounds such as polyphenols, carotenoids, sulphated polysaccharides and vitamins and have the potential to be used as ingredients in neutraceuticals. The antioxidant activity of crude 60% methanol extracts prepared from five Irish seaweeds, Ascophyllum nodosum, Laminaria hyperborea, Pelvetia canaliculata, Fucus vesiculosus and Fucus serratus were examined using in-vitro assays and a cell model system to determine the antioxidant activity of the extracts and their ability to protect against H2O2 and tert-BOOH-induced DNA damage and alterations in cellular antioxidant status in the human adenocarcinoma, Caco-2 cell line. To optimise the extraction of antioxidant compounds from seaweeds, an accelerated solvent extraction (ASE®) was used in combination with food grade solvents. The antioxidant activity of these extracts against H2O2 and tert-BOOH-induced DNA damage and alterations in cellular antioxidant status was also assessed. Extracts that exhibited the highest antioxidant activity, A. nodosum (100% water and 80% ethanol extracts) and F. vesiculosus (60% ethanol extract) were selected as ingredients for incorporation into fluid milk and yogurt at concentrations of 0.25% and 0.5%. The addition of the seaweed extracts to milk and yogurt did not affect the pH or shelf-life properties of the products. Seaweed addition did however significantly influence the colour properties of the milk and yogurt. Yellowness values were significantly higher in yogurts containing F. vesiculosus at both concentrations and A. nodosum (80% ethanol) at the 0.5% concentration. In milk, the F. vesiculosus (60% ethanol) and A. nodosum (80% ethanol) at both the 0.25% and the 0.5% concentrations had higher greenness and yellowness values than the milk containing A. nodosum (100% water). Sensory analysis revealed that appearance and flavour governed the overall acceptability of yogurts with the control yogurt, and yogurts containing A. nodosum (100% water) were the most preferred samples by panellists. However, in the milk trial the perception of a fishy taste was the determining factor in the negative perception of milk. The unsupplemented control and the milk containing A. nodosum (100% water) at a concentration of 0.5% were the most overall accepted milk samples by the sensory panellists. The antioxidant activity of the extracts in milk and yogurt remained stable during storage as determined by the in-vitro assays. Seaweed supplemented milk and yogurt were also subjected to an in-vitro digestion procedure which mimics the human digestive system. The milk and yogurt samples and their digestates were added to Caco-2 cells to investigate their antioxidant potential however neither the undigested or digested samples protected against H2O2-induced DNA damage in Caco-2 cells.

ANALYSIS OF IN VITRO AND IN VIVO ANTIOXIDANT PROPERTIES OF HYDROPHILIC FRACTIONS FROM THE SEAWEED HALIMEDA MONILE L.

In this work, in vitro and in vivo antioxidant properties of the marine algae Halimeda monile were assessed and the levels of some of its compounds likely to be responsible for such properties were determined. The estimated contents of total polyphenols, chlorophylls a and b and carotenoids were 179.5, 356.3, 452.8 and 42.2 mu g/g dry weight seaweed, respectively. The presence of terpenoids and flavonoids was also observed. The antioxidant activity of two polar fractions from H. monile (lyophilized aqueous extract and free phenolic acid fraction) was evaluated using three antioxidant assays: ferric reducing antioxidant power, 1,1-diphenyl-2-picrylhydrazyl and lipid peroxidation. Treatment of CCl4-induced liver damage in rats with extracts resulted in lower serum thiobarbituric acid-reactive substances levels and higher hepatic glutathione concentrations compared to those observed in the CCl4-treated group. Also, a significant increase in catalase activity was detected after treatment with the extracts. These results suggest that the seaweed H. monile could be a potential source for natural antioxidants.

Inhibition of LDL-oxidation and antioxidant properties related to polyphenol content of hydrophilic fractions from seaweed Halimeda Incrassata (Ellis) Lamouroux

LDL oxidation and oxidative stress are closely related to atherosclerosis. Therefore, natural antioxidants have been studied as promising candidates. In the present study, the LDL oxidation inhibition activity of bioactive compounds from Halimeda incrassata seaweed. associated to antioxidant capacity, was evaluated in vitro. Experimental work was conducted with lyophilized aqueous extract and phenolic-rich fractions of the seaweed and their effect on LDL oxidation was evaluated using heparin-precipitated LDL (hep-LDL) with exposure to Cu2+ ions and AAPH as the free radical generator. H. incrassata had a protective effect for hep-LDL in both systems and the presence of phenolic compounds contributed to the activity where phenolic-rich fractions showed significant capacity for inhibition of oxidation mediated by Cu2+ ions. The observed effect could be related to the antioxidant potential of polar fractions evidenced by reducing activity and DPPH center dot radical scavenging. The results obtained in vitro further support the antioxidant and LDL oxidation inhibition properties of H. incrassata and further knowledge toward future phytotherapeutic application of the seaweed.

Inhibition of LDL-oxidation and antioxidant properties related to polyphenol content of hydrophilic fractions from seaweed Halimeda Incrassata (Ellis) Lamouroux

LDL oxidation and oxidative stress are closely related to atherosclerosis. Therefore, natural antioxidants have been studied as promising candidates. In the present study, the LDL oxidation inhibition activity of bioactive compounds from Halimeda incrassata seaweed. associated to antioxidant capacity, was evaluated in vitro. Experimental work was conducted with lyophilized aqueous extract and phenolic-rich fractions of the seaweed and their effect on LDL oxidation was evaluated using heparin-precipitated LDL (hep-LDL) with exposure to Cu2+ ions and AAPH as the free radical generator. H. incrassata had a protective effect for hep-LDL in both systems and the presence of phenolic compounds contributed to the activity where phenolic-rich fractions showed significant capacity for inhibition of oxidation mediated by Cu2+ ions. The observed effect could be related to the antioxidant potential of polar fractions evidenced by reducing activity and DPPH radical scavenging. The results obtained in vitro further support the antioxidant and LDL oxidation inhibition properties of H. incrassata and further knowledge toward future phytotherapeutic application of the seaweed.

How to extract and expand randomness : a summary and explanation of existing results

We examine the use of randomness extraction and expansion in key agreement (KA) pro- tocols to generate uniformly random keys in the standard model. Although existing works provide the basic theorems necessary, they lack details or examples of appropriate cryptographic primitives and/or parameter sizes. This has lead to the large amount of min-entropy needed in the (non-uniform) shared secret being overlooked in proposals and efficiency comparisons of KA protocols. We therefore summa- rize existing work in the area and examine the security levels achieved with the use of various extractors and expanders for particular parameter sizes. The tables presented herein show that the shared secret needs a min-entropy of at least 292 bits (and even more with more realistic assumptions) to achieve an overall security level of 80 bits using the extractors and expanders we consider. The tables may be used to �nd the min-entropy required for various security levels and assumptions. We also �nd that when using the short exponent theorems of Gennaro et al., the short exponents may need to be much longer than they suggested.

Kava Anxiety Depression Spectrum Study (KADSS): a mixed methods RCT using an aqueous extract of Piper methysticum

Objectives: To report on the design, significance and potential impacts of the first documented human clinical trial assessing the anxiolytic and thymoleptic efficacy of an aqueous monoextract of Piper methysticum (kava). The significance of the qualitative element of our clinical trial is also explored. The Kava Anxiety Depression Spectrum Study (KADSS) is a 3-week placebocontrolled, double-blind, cross-over trial involving 60 adult participants (18—65) with elevated stable anxiety and varying levels of depressive symptoms. Aims: The aims of KADSS are: (1) to determine whether an aqueous standardised extract of kava is effective for the treatment of anxiety; (2) to assess the effects of kava on differing levels of depression; and (3) to explore participants’ experience of taking kava via qualitative research. The study also provides preliminary assessment of the safety of an aqueous extract of kava in humans. Conclusion: If results reveal that the aqueous kava preparation exerts significant anxiolytic effects and appears safe, potentially beneficial impacts may occur. Data supporting a safe and effective kava extract may encourage a re-introduction of kava to Europe, UK and Canada. This may provide a major socioeconomic benefit to Pacific Island nations, and to sufferers of anxiety disorders.

The Kava Anxiety Depression Spectrum Study (KADSS): A randomized, placebo-controlled, cross-over trial using an aqueous extract of Piper methysticum.

Rationale: Piper methysticum (Kava) has been withdrawn in European, British, and Canadian markets due to concerns over hepatotoxic reactions. The WHO recently recommended research into “aqueous” extracts of Kava. Objective: The objective of this study was to conduct the first documented human clinical trial assessing the anxiolytic and antidepressant efficacy of an aqueous extract of Kava. Design and participants: The Kava Anxiety Depression Spectrum Study was a 3-week placebo-controlled, double-blind crossover trial that recruited 60 adult participants with 1 month or more of elevated generalized anxiety. Five Kava tablets per day were prescribed containing 250 mg of kavalactones/day. Results: The aqueous extract of Kava reduced participants' Hamilton Anxiety Scale score in the first controlled phase by −9.9 (CI = 7.1, 12.7) vs. −0.8 (CI = −2.7, 4.3) for placebo and in the second controlled phase by −10.3 (CI = 5.8, 14.7) vs. +3.3 (CI = −6.8, 0.2). The pooled effect of Kava vs. placebo across phases was highly significant (p < 0.0001), with a substantial effect size (d = 2.24, η² [sub]p[sub] = 0.428). Pooled analyses also revealed highly significant relative reductions in Beck Anxiety Inventory and Montgomery–Asberg Depression Rating Scale scores. The aqueous extract was found to be safe, with no serious adverse effects and no clinical hepatotoxicity. Conclusions: The aqueous Kava preparation produced significant anxiolytic and antidepressant activity and raised no safety concerns at the dose and duration studied. Kava appears equally effective in cases where anxiety is accompanied by depression. This should encourage further study and consideration of globally reintroducing aqueous rootstock extracts of Kava for the management of anxiety.