Thesis (M.Sc. Agriculture (Plant Production)) -- University of Limpopo, 2014 / The management of root-knot nematodes (Meloidogyne spp.) has become a challenging task in tomato (Solanum lycorpesicum) production, due to the withdrawal of effective chemical nematicides. Currently, crude extracts of different plant species are being researched as alternative to chemical nematicides, with promising results. The objective of this study was to determine the effect of crude extracts of wild garlic (Tulbaghia violacea) on the growth of tomato under greenhouse conditions, and the suppression of M. incognita race 2 population densities. Treatments consisted of four levels of crude extracts viz. 0, 2, 4 and 8 g per pot, were arranged in a randomised complete block design with 10 replicates. Seedlings were inoculated with 1000 juveniles of M. incognita race 2 at transplanting and treated with crude extracts two days later. At 56 days, the crude extract of T. violacea increased plant height, stem diameter, number of cluster, flowers, fruits and leaves by 43-73%, 108-200%, 57-81%, 55-110%, 170-223% and 51-66%, respectively. It also increased the root mass and shoot mass by 95% and 96%, respectively. Crude extracts of T. violacea did not have any effect on soil pH and electrical conductivity (EC). Crude extracts of T. violacea consistently reduced population densities of M. incognita race 2 by 50, 64 and 73% in roots at 2, 4 and 8 g crude extracts, respectively and by 21, 30 and 58% in soil at similar levels, respectively. In conclusion, crude extracts of T. violacea have the potential to improve growth of tomato plants and suppress population densities of M. incognita race 2 and could be used as botanical nematicide in tomato production.

<p>Cilj ove doktorske disertacije je bio odrediti niz tehnolo&scaron;kih postupaka koji bi omogućili iskori&scaron;ćenje sremu&scaron;a u prehrambene i farmaceutske svrhe u svežem ili suvom stanju, kao biljne droge za pripremu različitih formi ekstrakata bogatih bioaktivnim jedinjenjima izolovanih iz njega.<br />List sremu&scaron;a bere se u vrlo kratkom vremenskom periodu tokom proleća, a nakon branja njegovi listovi vrlo brzo venu i gube svoja senzorna, nutritivna i funkcionalna svojstva, zbog čega je dostupnost ove biljne sirovine vremenski vrlo ograničena. Zbog toga su ispitani uslovi skladi&scaron;tenja svežeg lista sremu&scaron;a kako bi se sagledala trajnost sremu&scaron;a za konzumiranje u svežem stanju, ali i mogućnost primene skladi&scaron;tenog sremu&scaron;a za ekstrakciju bioaktivnih jedinjenja. Za dobijanje osu&scaron;enog lista sremu&scaron;a ispitan je uticaj različitih tehnika su&scaron;enja (konvektivno i vakuumsko su&scaron;enje) i temperature su&scaron;enja (40, 50, 60 i 70 &deg;C) na sadržaj bioaktivnih komponenti u osu&scaron;enom proizvodu. Primenom odgovarajuće tehnike su&scaron;enja, povećava se stabilnost i dostupnost listova sremu&scaron;a za potrebe prehrambene i farmaceutske industrije. Primenom savremenih ekstrakcionih tehnika kao &scaron;to je ekstrakcije superkritičnim ugljen-dioksidom, ekstrakcije subkritičnom vodom i ultrazvučne ekstrakcije, ispitan je uticaj procesnih parametara različitih ekstrakcionih tehnika, poput ultrazvučne ekstrakcije (temperatura, vreme ekstrakcije, snaga ultrazvuka, uticaj rastvarača) i ekstrakcije subkritičnom vodom (temperatura, vreme ekstrakcije, dodatak modifikatora), superkritične ekstrakcije ugljen-dioksidom (pritisak i temperatura), dok je maceracija kao konvencionalna tehnika ekstrakcije primenjena za poređenje efikasnosti modernih ekstrakcionih postupaka. U dobijenim ekstraktima ispitan je sadržaj polifenolnih komponenti, antioksidantna aktivnost i sadržaj sumpornih jedinjenja u cilju optimizacije ekstrakcionih parametara i radi utvrđivanja uslova ekstrakcije pri kojima se postiže najveći kvalitet ekstrakata u pogledu sadržaja ciljanih bioaktivnih komponenti.&nbsp; Za odabrane ekstrakte ispitan je antimikrobni potencijal na različite Gram pozitivne i Gram negativne bakterijske sojeve. Za dobijanje stabilnijih formi odabranih ekstrakata primenjena je enkpasulacija spray drying i spray congeling tehnikom. Enkapsuliranim ekstraktima su određene fizičko-hemijske osobine (hemijski sastav, sadržaj vlage, higroskopnost, moć rehidratacije, WAI i WSI) i biolo&scaron;ka aktivnost (antimikrobna i antioksidativna aktivnost).</p> / <p>The aim of this doctoral dissertation was to determine various technological procedures that would enable the use of fresh and dryed wild garlic leaves in foods and pharmaceutical industry, as a herbal drug or nutraceuticals.<br />The availability period of fresh wild garlic is very short and only during the spring season. After harvest, the leaves are quite perishable, wither very quickly and lose their sensory, nutritive and functional properties. This makes the availability time of this herbal raw material very limited. Therefore, the conditions for storing the fresh leaves of wild garlic and the influences of storing conditions on composition of bioactive compounds were examined in order to extend their use in fresh state. In order to obtain dry herbal drug for extraction, influence of various drying techniques (convection and vacuum drying) and drying temperatures (40, 50, 60, 70 &deg;C) were examinated. The stability and availability of wild garlic in food and pharmaceutical industries was extended by drying. The influence of process parameters of different modern extraction techniques - ultrasonic extraction (temperature, extraction time, ultrasonic power, solvent effect) and subcritical water extraction , extraction time, modifier addition), supercritical carbon dioxide extraction (pressure and temperature) - was examined, while maceration (as a conventional extraction technique) was used to evaluate the efficiency of modern extraction methods. The obtained extracts were analysed in terms of the polyphenolic content, the antioxidant activity and the content of sulfur compounds in order to optimize the extraction parameters and determine the extraction conditions for achieving the highest quality of extracts in terms of the content of the target bioactive components. Antimicrobial potential for different Gram positive and Gram negative bacterial strains was tested for selected<br />extracts. To obtain more stable forms of selected extracts encapsulation techniques - spray drying and spray congeling - were applied. Encapsulated extracts were evaluated in terms of physico-chemical properties (chemical composition, moisture content, hygroscopicity, rehydration power, WAI and WSI) and biological activity (antimicrobial and antioxidant activity).</p>

Allium sativum has a long tradition in medicine. While much is known about its potential healthy effects, nearly nothing is known about wild garlic (allium sativum, ramson), which is very common in the area of Leipzig and has been used as a herbal remedy since centuries. The goal of the present study was to assess a potential anti-platelet activity of these two allium species and to try to identify the chemical active principle. For that purpose various extracts (hydrophilic and lipophilic) were prepared from Allium sativum and Allium ursinum, and analysed using thin layer chromatography and HPLC. After identifying an active, i.e. antiaggregatory extract (see below), this was fractionated and the active fraction was further sub-fractionated for subsequent chemical analysis by mass spectroscopy, ESI (Electrospray ionization), and COSY (Correlation effect spectroscopy), and functional testing. Anti-platelet activity was assessed in human platelets (platelet rich plasma) using a classical turbidimetric method. Platelets were stimulated with various agonists (arachidonic acid, ADP, epinephrine, collagen, A23187) with and without the addition of the extracts or the fractions /sub-fractions. Both Allium Ursinum and Allium sativum extracts exert antiaggregatory effects with EC50 values around 0.1 mg/ml. The garlic extracts are acting by inhibition of the ADP pathway comparable as known from the clinically used drug clopidegrol.The pharmacological active antiaggregatory component of the extracts appears to be lipophilic rather than hydrophilic. This is the first report on an antiplatelet activity of Allium Ursinum. One final structure determined by HPLC, MS, ESI and COSY which exerts the antiplatelets inhibitory effect is β-sitosterol-3-O-β-D-glucoside of the fraction 7-14 crystals. It is considered that about three up to five grams of dried leaves might be enough to exert antiaggregatory effects (comment: in pharmacy normally dried plant material is used in therapy). The second compound with antiaggregatory activity was identified as 1-β-D-galactopyranoside-2, 3-bis-linolenic glycerate. The problem of loosing the active volatile oily components by drying the leaves in future studies looking for the clinical use may be solved by looking for a raw or a refined extract which would be the form of a real phytomedical drug; for example capsules about 120 to 200 micrograms of an alcoholic or better an heptane / oily extract gained from wood garlic leaves would be an useful drug formulation to reach respective concentrations in blood. However, we have to admit that since our investigations were in-vitro, the in-vivo situation is somewhat different due to the metabolism, which is nearly unknown. Nevertheless, this study shows for the first time that allium ursinum does exert anti-platelet activity and that both allium species can unfold antiaggregatory effects which are worth to be investigated in subsequent in-vivo studies. β-sitosterol-3-O-β-D-glucoside and 1-β-D-galactopyranoside-2, 3-bis-linolenic glycerate could be identified as active antiaggregatory principals.

Allium sativum has a long tradition in medicine. While much is known about its potential healthy effects, nearly nothing is known about wild garlic (allium sativum, ramson), which is very common in the area of Leipzig and has been used as a herbal remedy since centuries. The goal of the present study was to assess a potential anti-platelet activity of these two allium species and to try to identify the chemical active principle. For that purpose various extracts (hydrophilic and lipophilic) were prepared from Allium sativum and Allium ursinum, and analysed using thin layer chromatography and HPLC. After identifying an active, i.e. antiaggregatory extract (see below), this was fractionated and the active fraction was further sub-fractionated for subsequent chemical analysis by mass spectroscopy, ESI (Electrospray ionization), and COSY (Correlation effect spectroscopy), and functional testing. Anti-platelet activity was assessed in human platelets (platelet rich plasma) using a classical turbidimetric method. Platelets were stimulated with various agonists (arachidonic acid, ADP, epinephrine, collagen, A23187) with and without the addition of the extracts or the fractions /sub-fractions. Both Allium Ursinum and Allium sativum extracts exert antiaggregatory effects with EC50 values around 0.1 mg/ml. The garlic extracts are acting by inhibition of the ADP pathway comparable as known from the clinically used drug clopidegrol.The pharmacological active antiaggregatory component of the extracts appears to be lipophilic rather than hydrophilic. This is the first report on an antiplatelet activity of Allium Ursinum. One final structure determined by HPLC, MS, ESI and COSY which exerts the antiplatelets inhibitory effect is β-sitosterol-3-O-β-D-glucoside of the fraction 7-14 crystals. It is considered that about three up to five grams of dried leaves might be enough to exert antiaggregatory effects (comment: in pharmacy normally dried plant material is used in therapy). The second compound with antiaggregatory activity was identified as 1-β-D-galactopyranoside-2, 3-bis-linolenic glycerate. The problem of loosing the active volatile oily components by drying the leaves in future studies looking for the clinical use may be solved by looking for a raw or a refined extract which would be the form of a real phytomedical drug; for example capsules about 120 to 200 micrograms of an alcoholic or better an heptane / oily extract gained from wood garlic leaves would be an useful drug formulation to reach respective concentrations in blood. However, we have to admit that since our investigations were in-vitro, the in-vivo situation is somewhat different due to the metabolism, which is nearly unknown. Nevertheless, this study shows for the first time that allium ursinum does exert anti-platelet activity and that both allium species can unfold antiaggregatory effects which are worth to be investigated in subsequent in-vivo studies. β-sitosterol-3-O-β-D-glucoside and 1-β-D-galactopyranoside-2, 3-bis-linolenic glycerate could be identified as active antiaggregatory principals.