Direct anthelmintic effects of condensed tannins from diverse plant sources against Ascaris suum

Ascaris suum is one of the most prevalent nematode parasites in pigs and causes significant economic losses, and also serves as a good model for A. lumbricoides, the large roundworm of humans that is ubiquitous in developing countries and causes malnutrition, stunted growth and compromises immunity to other pathogens. New treatment options for Ascaris infections are urgently needed, to reduce reliance on the limited number of synthetic anthelmintic drugs. In areas where Ascaris infections are common, ethno-pharmacological practices such as treatment with natural plant extracts are still widely employed. However, scientific validation of these practices and identification of the active compounds are lacking, although observed effects are often ascribed to plant secondary metabolites such as tannins. Here, we extracted, purified and characterised a wide range of condensed tannins from diverse plant sources and investigated anthelmintic effects against A. suum in vitro. We show that condensed tannins can have potent, direct anthelmintic effects against A. suum, as evidenced by reduced migratory ability of newly hatched third-stage larvae and reduced motility and survival of fourth-stage larvae recovered from pigs. Transmission electron microscopy showed that CT caused significant damage to the cuticle and digestive tissues of the larvae. Furthermore, we provide evidence that the strength of the anthelmintic effect is related to the polymer size of the tannin molecule. Moreover, the identity of the monomeric structural units of tannin polymers may also have an influence as gallocatechin and epigallocatechin monomers exerted significant anthelmintic activity whereas catechin and epicatechin monomers did not. Therefore, our results clearly document direct anthelmintic effects of condensed tannins against Ascaris and encourage further in vivo investigation to determine optimal strategies for the use of these plant compounds for the prevention and/or treatment of ascariosis.

Assessment of the anthelmintic activity of medicinal plant extracts and purified condensed tannins against free-living and parasitic stages of Oesophagostomum dentatum

Background: Plant-derived condensed tannins (CT) show promise as a complementary option to treat gastrointestinal helminth infections, thus reducing reliance on synthetic anthelmintic drugs. Most studies on the anthelmintic effects of CT have been conducted on parasites of ruminant livestock. Oesophagostomum dentatum is an economically important parasite of pigs, as well as serving as a useful laboratory model of helminth parasites due to the ability to culture it in vitro for long periods through several life-cycle stages. Here, we investigated the anthelmintic effects of CT on multiple life-cycles stages of O. dentatum. Methods: Extracts and purified fractions were prepared from five plants containing CT and analysed by HPLC-MS. Anthelmintic activity was assessed at five different stages of the O. dentatum life cycle; the development of eggs to infective third-stage larvae (L3), the parasitic L3 stage, the moult from L3 to fourth-stage larvae (L4), the L4 stage and the adult stage. Results: Free-living larvae of O. dentatum were highly susceptible to all five plant extracts. In contrast, only two of the five extracts had activity against L3, as evidenced by migration inhibition assays, whilst three of the five extracts inhibited the moulting of L3 to L4. All five extracts reduced the motility of L4, and the motility of adult worms exposed to a CT-rich extract derived from hazelnut skins was strongly inhibited, with electron microscopy demonstrating direct damage to the worm cuticle and hypodermis. Purified CT fractions retained anthelmintic activity, and depletion of CT from extracts by pre-incubation in polyvinylpolypyrrolidone removed anthelmintic effects, strongly suggesting CT as the active molecules. Conclusions: These results suggest that CT may have promise as an alternative parasite control option for O. dentatum in pigs, particularly against adult stages. Moreover, our results demonstrate a varied susceptibility of different life-cycle stages of the same parasite to CT, which may offer an insight into the anthelmintic mechanisms of these commonly found plant compounds.

Anthelmintic activity of trans-cinnamaldehyde and A- and B-type proanthocyanidins derived from cinnamon (Cinnamomum verum)

Cinnamon (Cinnamomum verum) has been shown to have anti-inflammatory and antimicrobial properties, but effects on parasitic worms of the intestine have not been investigated. Here, extracts of cinnamon bark were shown to have potent in vitro anthelmintic properties against the swine nematode Ascaris suum. Analysis of the extract revealed high concentrations of proanthocyanidins (PAC) and trans-cinnamaldehyde (CA). The PAC were subjected to thiolysis and HPLC-MS analysis which demonstrated that they were exclusively procyanidins, had a mean degree of polymerization of 5.2 and 21% of their inter-flavan-3-ol links were A-type linkages. Purification of the PAC revealed that whilst they had activity against A. suum, most of the potency of the extract derived from CA. Trichuris suis and Oesophagostomum dentatum larvae were similarly susceptible to CA. To test whether CA could reduce A. suum infection in pigs in vivo, CA was administered daily in the diet or as a targeted, encapsulated dose. However, infection was not significantly reduced. It is proposed that the rapid absorption or metabolism of CA in vivo may prevent it from being present in sufficient concentrations in situ to exert efficacy. Therefore, further work should focus on whether formulation of CA can enhance its activity against internal parasites.

Impact of chemical structure of flavanol monomers and condensed tannins on in vitro anthelmintic activity against bovine nematodes

Plants containing condensed tannins (CT) may have potential to control gastrointestinal nematodes (GIN) of cattle. The aim was to investigate the anthelmintic activities of four flavan-3-ols, two galloyl derivatives and 14 purified CT fractions, and to define which structural features of CT determine the anti-parasitic effects against the main cattle nematodes. We used in vitro tests targeting L1 larvae (feeding inhibition assay) and adults (motility assay) of Ostertagia ostertagi and Cooperia oncophora. In the larval feeding inhibition assay, O. ostertagi L1 were significantly more susceptible to all CT fractions than C. oncophora L1. The mean degree of polymerization of CT (i.e. average size) was the most important structural parameter: large CT reduced larval feeding more than small CT. The flavan-3-ols of prodelphinidin (PD)-type tannins had a stronger negative influence on parasite activity than the stereochemistry, i.e. cis- vs trans-configurations, or the presence of a gallate group. In contrast, for C. oncophora high reductions in the motility of larvae and adult worms were strongly related with a higher percentage of PDs within the CT fractions while there was no effect of size. Overall, the size and the percentage of PDs within CT seemed to be the most important parameters that influence anti-parasitic activity.

Galloylated proanthocyanidins from shea (Vitellaria paradoxa) meal have potent anthelmintic activity against Ascaris suum

Proanthocyanidins (PA) from shea (Vitellaria paradoxa) meal were investigated by thiolytic degradation with benzyl mercaptan and the reaction products were analysed by high performance liquid chromatography–mass spectrometry. These PA were galloylated (≈40%), contained only B-type linkages and had a high proportion of prodelphinidins (>70%). The mean degree of polymerisation was 8 (i.e. average molecular size was 2384 Da) and epigallocatechin gallate (EGCg) was the major flavan-3-ol subunit in PA. Shea meal also proved to be a potentially valuable source for extracting free flavan-3-ol-O-gallates, especially EGCg (575 mg/kg meal), which is known for its health and anti-parasitic benefits. Proanthocyanidins were isolated and tested for bioactivity against Ascaris suum, which is an important parasite of pigs. Migration and motility tests revealed that these PA have potent activity against this parasitic nematode.

Potent anthelmintic activity of galloylated proanthocyanidins from shea meal (Vitellaria paradoxa)

Proanthocyanindins (PAs) from shea meal (SM), a by-product obtained after lipid extraction of the nuts, contained B-type linkages, had a high ratio of prodelphinidins (73%) and were galloylated (42%). The average polymer size was 8 flavan-3-ol subunits (≈2384 Daltons) and epigallocatechin gallate was the major subunit. Purified PA fractions from SM were tested in vitro for anthelmintic properties against gastrointestinal nematodes from ruminants (H. contortus and T. colubriformis) [1] by the larval exsheathment inhibition assay and from pigs (A. suum) by the larval migration inhibition assay. Results showed that PAs from SM have a potent anthelmintic activity against those parasites similar to white clover (Trifolium repens) flowers (WCF) [1, 2] (EC50 µg/mL; SM: 55.1, 16.5, 75.9; WCF: 37.4, 14.5, 110.1 for A. suum, H. contortus and T. colubriformis respectively). WCF PAs are constituted almost exclusively of prodelphinidin (PD) compared to SM (98% vs. 73%) but do not contained galloylated PAs. Studies [1, 2] have shown that anthelmintic activity of PAs was mainly associated with their PD ratio but our current results suggest that galloylation can be a major factor to anthelmintic activity and SM as a potential nutraceutical anthelmintic feed for controlling parasitic nematodes.

Polymerization-dependent activation of porcine γδ T-cells by proanthocyanidins

Plant-derived proanthocyanidins (PAC) have been promoted as a natural method of improving health and immune function in livestock. It has previously been shown that PAC are effective agonists for activating ruminant γδ T-cells in vitro, however effects on other livestock species are not yet clear. Moreover, the fine structural characteristics of the PAC which contribute to this stimulatory effect have not been elucidated. Here, we demonstrate activation of porcine γδ T-cells by PAC via up-regulation of CD25 (IL-2Rα) and show that 1) activation is dependent on degree of polymerization (DP), with PAC fractions containing polymers with mean DP >6 significantly more effective than fractions with mean DP <6, whilst flavan-3-ol monomers (the constituent monomeric units of PAC) did not induce CD25 expression and 2) both procyanidin and prodelphinidin-type PAC are effective agonists. Furthermore, we show that this effect of PAC is restricted to the γδ T-cell population within porcine peripheral mononuclear cells as significant CD25 up-regulation was not observed in non γδ T-cells, and no activation (via CD80/86 up-regulation) was evident in monocytes. Our results show that dietary PAC may contribute to enhancement of innate immunity in swine via activation of γδ T-cells.

Förpackningsframställning för skönhetsvård

This degree project was intendet to construct and design packaging for beauty care. At the requestof AssiDomän Frövi the project group were instructed to innovate creative packagesmade of their cardboard Frövi Bright.Three different packages have been developed. Two of them are for child care products and onepackage is for several bath products. These results show that there are possibilities to create creativeand different packages of the cardboard Frövi Bright.The report also enters deeply into the special qualities of Frövi Bright and studies of packagingdesign.

Implementering av felpredikteringoch frekvenshopp i ett 2,4 GHzradiosystem

Maxicap AB är ett företag som utvecklar och tillverkar elektronisk utrustning för bland annat olika mätuppgifter. Målet med detta examensjobb var att undersöka och implementera en stabil trådlös dataöverföringsmetod med den utrustning som Maxicap ville använda. Detta skulle åstadkommas/testas genom att sända temperatursensorvärden trådlöst med Nordic Semiconductor’ssändtagare (eng. transceiver) styrda av Atmel’s mikrokontrollers. Arbetet fokuserade på att få till denna stabila trådlösa överföring genom att implementera frekvenshoppning och datapaketskontroll på datatrafiken inom det fria 2,4 GHz ISM-bandet. All den C-kod som utvecklades i detta arbete är skriven för att vara lättanpassad till flera olika modeller av mikrokontrollers inom Atmel’s produktserie samt att kunna användas till andra liknande projekt. Koden är väldokumenterad med kommentarer vid varje funktionsblock som beskriver vad funktionerna gör och hur de skall användas. Arbetet resulterade i två fungerande prototyper. En enhet känner av temperaturen i luften och skickar sedan detta temperaturvärde trådlöst till enhet nummer två. Detta värde samt några inställningsvärden för mottagaren visas sedan på den enhetens display.

Reflection of matter waves in potential structures

In this paper the behavior of matter waves in suddenly terminated potential structures is investigated numerically. It is shown that there is no difference between a fully quantum mechanical treatment and a semiclassical one with regards to energy redistribution. For the quantum case it is demonstrated that there can be substantial reflection at the termination. The neglect of backscattering by the semiclassical method brings about major differences in the case of low kinetic energies. A simple phenomenological model is shown to partially explain the observed backscattering using dynamics of reduced dimensionality.

Adiabatic propagation in potential structures

In this work the adiabatic approximation is applied to the propagation of matter waves in confined geometries like those experimentally realized in recent atom optical experiments. Adiabatic propagation along a channel is assumed not to mix the various transverse modes. Nonadiabatic corrections arise from the potential squeezing and bending. Here we investigate the effect of the former. Detailed calculations of two-dimensional propagation are carried out both exactly and in an adiabatic approximation. This offers the possibility to analyze the validity of adiabaticity criteria. A semiclassical (sc) approach, based on the sc Massey parameter is shown to be inadequate, and the diffraction due to wave effects must be included separately. This brings in the Fresnel parameter well known from optical systems. Using these two parameters, we have an adequate understanding of adiabaticity on the system analyzed. Thus quantum adiabaticity must also take cognizance of the intrinsic diffraction of matter waves.

Localization in splitting of matter waves

In this paper we present an analysis of how matter waves, guided as propagating modes in potential structures, are split under adiabatic conditions. The description is formulated in terms of localized states obtained through a unitary transformation acting on the mode functions. The mathematical framework results in coupled propagation equations that are decoupled in the asymptotic regions as well before as after the split. The resulting states have the advantage of describing propagation in situations, for instance matter-wave interferometers, where local perturbations make the transverse modes of the guiding potential unsuitable as a basis. The different regimes of validity of adiabatic propagation schemes based on localized versus delocalized basis states are also outlined. Nontrivial dynamics for superposition states propagating through split potential structures is investigated through numerical simulations. For superposition states the influence of longitudinal wave-packet extension on the localization is investigated and shown to be accurately described in quantitative terms using the adiabatic formulations presented here.