Triterpenes from Minquartia guianensis (Olacaceae) and in vitro antimalarial activity

Minquartia guianensis, popularly known as acariquara, was phytochemically investigated. The following triterpenes were isolated from the dichloromethane extract of leaves: lupen-3-one (1), taraxer-3-one (2) and oleanolic acid (3). The dichloromethane extract of branches yielded the triterpene 3β-methoxy-lup-20(29)-ene (4). The chemical structures were characterized by NMR data. Plant extracts, substance 3, squalene (5) and taraxerol (6), (5 and 6 previously isolated), were evaluated by in vitro assay against chloroquine resistant Plasmodium falciparum. The dichloromethane extract of leaves and the three triterpenes assayed have shown partial activity. Thus, these results demonstrated that new potential antimalarial natural products can be found even in partially active extracts.

Use of raw Euphorbia tirucalli extract for inhibition of ascitic Ehrlich tumor

Objective: to evaluate the effect of the Euphorbia tirucalli hydroalcoholic extract (ETHE) on the development of Ehrlich Tumor, in its ascitic form. Methods: we intraperitoneally inoculated 15 Swiss mice with 10.44 x 107 cells of Ehrlich Tumor and divided them in two groups one day after: ETHE Group (eight mice), treated with a dosage of 125 mg/kg/day of EHTE for five days; and Control Group (seven mice), treated only with 0.9% isotonic saline solution over the same period. The treatment was done by gavage. Ten days after inoculation, four mice from each group were sacrificed for quantification of tumor cell number, ascitic fluid volume and bone marrow cell number. The remaining animals were maintained to evaluate survival. Results: The ascitic fluid volume and the tumor cell number were decreased in the ETHE group when compared with the control group, but with no statistical significance. On the other hand, survival was higher in the ETHE group, as well as the number of bone marrow cells. Conclusion: Treatment with ETHE after inoculation of Ehrlich Tumor decreases its development and increases survival and the bone marrow cellularity, thus reducing the myelosuppression present in the Ehrlich Tumor bearing mice.

Allelopathic effect of black mustard tissues and root exudates on some crops and weeds

Laboratory and greenhouse experiments were conducted to evaluate the phytotoxic effect of black mustard extracts and root exudates on two crops: Trifolium alexandrinum and Triticum aestivum, and two weeds: Phalaris paradoxa and Sisymbrium irio. The seeds were treated with aqueous and ethanolic extracts and chloroform for eight days, or subjected to root exudates of just harvested mustard in a greenhouse for five weeks. High-performance liquid chromatography (HPLC) was used to quantify phytotoxins from plant tissues. Seed germination of P. paradoxa was reduced with the lowest concentration of the different extracts. However, the aqueous extract at 4% completely curtailed the germination of all the target species. In general, plant extracts had a concentration-dependent reduction of seedling growth of the target species. However, the ethanolic extract, at the lowest concentration, has stimulated the shoot length of both T. alexandrinum and T. aestivum, and the root length of the former. Mustard root exudates inhibited emergence and growth of the target species throughout the experiment. Ferulic and syringic acids were the dominant allelochemicals found when HPLC was used.

Allelopathic effects of Rhynchosia capitata on germination and seedling growth of mungbean

Experiments were conducted to evaluate the allelopathic influence of Rhynchosia capitata on germination and seedling growth of mungbean (Vigna radiate) along with identification of the phytotoxic substances responsible for this activity. Water extracts of root, shoot, leaf, fruit and whole plant were prepared by soaking them in water in a ratio of 1:20 (w/v) for 24 h. All the extracts affected germination and seedling growth of mungbean, but higher inhibition was seen with R. capitata leaf water extracts. A linear decrease in the germination characteristics of mungbean was observed with the decrease in the concentration of leaf extract from 5% to 1%. The soil-incorporated residues (1-4% w/w) of R. capitata stimulated the growth of root and hypocotyl at low concentrations, while it inhibited their growth at higher concentrations. Rhynchosia capitata soil-incorporated residues (4% w/w) significantly reduced the seedling vigour index of mungbean in addition to their significant effect on total germination. A significant amount of water-soluble phenolic acids were found in R. capitata plant extracts. The content of total phenolic acids was higher in the leaf extract compared to that of the stem, fruit or root extracts. Two phenolic acids including vanillic acid and 4‑(hydroxymethyl) benzoic acid were found in R. capitata leaf extracts.

Allelopathic action of parthenium and its rhizospheric soil on maize as influenced by growing conditions

Biosynthesis and subsequent release of allelochemicals by a plant into the environment is supposed to be influenced by its growing conditions. To ascertain what will be the allelopathic action of plant parts and rhizospheric soils of parthenium (Parthenium hysterophorus) growing at various farm locations with varied growing conditions, germination and seedling growth of maize hybrid (DK 6142) were assayed by sowing its seeds in petri plates lined with filter paper and pots filled with soil. Minimum germination percentage (30.0%), germination index (2.01), germination energy (36.3), seedling length (3.3 cm), seedling biomass (10 mg) and seedling vigor index (99.0) of maize were observed in leaf extract followed by fruit and whole plant extracts of parthenium growing near the field border. Rhizospheric soil collected underneath parthenium growing near a water channel caused maximum reductions in germination index (30.8%), germination energy (40.6%), seedling length (32.6%), seedling biomass (35.1%) and seedling vigor index (34.3%) of maize compared with that soil without any vegetation. Phytotoxic inhibitory effects of both parthenium plant and rhizospheric soil were more pronounced on maize root than its shoot growth. The higher suppressive action against germination and seedling growth of maize was probably due to higher total phenolic concentrations (6678.2 and 2549.0 mg L-1) and presence of phenolic compounds viz., gallic, caffeic, 4-hydroxy-3-methoxy benzoic, p-coumaric and m-coumaric acids; and ferulic, vanillic, syringic and m-coumaric acids in aqueous leaf extract of parthenium uprooted near the field border and its rhizospheric soil collected near a water channel, respectively.

Estimation of weed dry biomass and grain yield as a function of growth and yield traits under allelopathic weed management in maize

Growing concerns about toxicity and development of resistance against synthetic herbicides have demanded looking for alternative weed management approaches. Allelopathy has gained sufficient support and potential for sustainable weed management. Aqueous extracts of six plant species (sunflower, rice, mulberry, maize, brassica and sorghum) in different combinations alone or in mixture with 75% reduced dose of herbicides were evaluated for two consecutive years under field conditions. A weedy check and S-metolachlor with atrazine (pre emergence) and atrazine alone (post emergence) at recommended rates was included for comparison. Weed dynamics, maize growth indices and yield estimation were done by following standard procedures. All aqueous plant extract combinations suppressed weed growth and biomass. Moreover, the suppressive effect was more pronounced when aqueous plant extracts were supplemented with reduced doses of herbicides. Brassica-sunflower-sorghum combination suppressed weeds by 74-80, 78-70, 65-68% during both years of study that was similar with S-metolachlor along half dose of atrazine and full dose of atrazine alone. Crop growth rate and dry matter accumulation attained peak values of 32.68 and 1,502 g m-2 d-1 for brassica-sunflower-sorghum combination at 60 and 75 days after sowing. Curve fitting regression for growth and yield traits predicted strong positive correlation to grain yield and negative correlation to weed dry biomass under allelopathic weed management in maize crop.

Interference of propylene glycol with the hole-board test

Experimental drugs and/or plant extracts are often dissolved in solvents, including propylene glycol. Nevertheless, there is evidence for psychoactive properties of this alcohol. In this study we found that in the hole-board test 10% propylene glycol did not modify the head-dipping behavior. However, 30% propylene glycol induced an increase in the number of head-dips (46.92 ± 2.37 compared to 33.83 ± 4.39, P<0.05, ANOVA/Student-Newman-Keuls), an effect comparable to that obtained with 0.5 mg/kg diazepam (from 33.83 ± 4.39 to 54 ± 3.8, P<0.01, ANOVA/Student-Newman-Keuls). These results demonstrate that 30% propylene glycol has significant anxiolytic effects in this model and therefore cannot be used as an innocuous solvent.

Antiprotozoal and molluscicidal activities of five Brazilian plants

Leishmaniasis, Chagas' disease and schistosomiasis (bilharzia) are parasitic diseases with wide distribution on the American continent, affecting millions of people. In the present study, biological assays for antiprotozoal and molluscicidal activities were carried out with ethanolic extracts of plant species from the Brazilian part of the Upper Paraná River. Crude extracts were obtained by percolation with absolute ethanol from the leaves of Cayaponia podantha Cogn., Nectandra falcifolia (Nees) Castiglioni and Paullinia elegans Cambess., as well as from the aerial parts of Helicteres gardneriana St. Hil. & Naud. and Melochia arenosa Benth., all belonging to genera used in folk medicine. Trypanocidal activity of plants was assayed on epimastigote cultures in liver infusion tryptose. Anti-leishmanial activity was determined over cultures of promastigote forms of the parasite in Schneider's Drosophila medium. Microscopic countings of parasites, after their incubation in the presence of different concentrations of the crude extracts, were made in order to determine the percentage of growth inhibition. C. podantha and M. arenosa, at a concentration of 10 µg/mL, showed 90.4 ± 11.52 and 88.9 ± 2.20% growth inhibition, respectively, of epimastigote forms of Trypanosoma cruzi, whereas N. falcifolia demonstrated an LD50 of 138.5 µg/mL against promastigote forms of Leishmania (Viannia) braziliensis. Regarding molluscicidal activity, the acute toxicity of the extracts on Biomphalaria glabrata was evaluated by a rapid screening procedure. M. arenosa was 100% lethal to snails at 200 µg/mL and showed an LD50 of 143 µg/mL. Screening of plant extracts represents a continuous effort to find new antiparasitic drugs.

The mutagenic and antimutagenic effects of the traditional phytoestrogen-rich herbs, Pueraria mirifica and Pueraria lobata

Pueraria mirifica is a Thai phytoestrogen-rich herb traditionally used for the treatment of menopausal symptoms. Pueraria lobata is also a phytoestrogen-rich herb traditionally used in Japan, Korea and China for the treatment of hypertension and alcoholism. We evaluated the mutagenic and antimutagenic activity of the two plant extracts using the Ames test preincubation method plus or minus the rat liver mixture S9 for metabolic activation using Salmonella typhimurium strains TA98 and TA100 as indicator strains. The cytotoxicity of the two extracts to the two S. typhimurium indicators was evaluated before the mutagenic and antimutagenic tests. Both extracts at a final concentration of 2.5, 5, 10, or 20 mg/plate exhibited only mild cytotoxic effects. The plant extracts at the concentrations of 2.5, 5 and 10 mg/plate in the presence and absence of the S9 mixture were negative in the mutagenic Ames test. In contrast, both extracts were positive in the antimutagenic Ames test towards either one or both of the tested mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide and benzo(a)pyrene. The absence of mutagenic and the presence of anti-mutagenic activities of the two plant extracts were confirmed in rec-assays and further supported by a micronucleus test where both plant extracts at doses up to 300 mg/kg body weight (equivalent to 16 g/kg body weight plant tuberous powder) failed to exhibit significant micronucleus formation in rats. The tests confirmed the non-mutagenic but reasonably antimutagenic activities of the two plant extracts, supporting their current use as safe dietary supplements and cosmetics.

Inhibition de l’absorption intestinale du glucose par les produits naturels issus de la pharmacopée traditionnelle des Cris de la Baie James

Le diabète de type 2 et l'obésité sont des problèmes de santé majeurs et les peuples autochtones sont particulièrement à risque. Pour remédier à ce problème largement répandu dans les populations autochtones canadiennes pour qui la médication moderne n’est pas culturellement adaptée, notre équipe s’est donné comme objectif d’étudier les activités potentielles antidiabétique et anti-obésité de la pharmacopée traditionnelle des Cris de la Baie James. Le but de cette étude est de tester l’hypothèse selon laquelle certaines plantes médicinales pourraient inhiber l'absorption intestinale du glucose, une activité anti-hyperglycémique qui, par la même occasion, contribuerait à combattre l’obésité. Les extraits éthanoliques de dix-sept plantes médicinales de la forêt boréale ont été testés dans des cellules intestinales Caco-2 et comparés à l’effet d’inhibiteurs compétitifs connus, tels que la phlorizine et la phlorétine. Ces inhibiteurs sont des composés polyphénoliques qui partagent de nombreuses caractéristiques structurelles avec des constituants moléculaires de plusieurs plantes Cri. Les résultats démontrent que treize des dix-sept extraits de plantes ont inhibé de façon significative l'absorption intestinale du 3H-D-glucose. Pour valider ces effets in vivo, quatre extraits ont été administrés à des rats Wistar par gavage intragastrique (250 mg/kg) en même temps qu’un bolus de glucose (3 g/kg). Suite à ce gavage, deux de ces extraits ont restreint l’augmentation de la glycémie d'environ 40% par rapport à un contrôle sans extrait. Ces résultats indiquent qu’une inhibition compétitive de l'absorption intestinale du glucose peut être atteinte par des extraits bruts de plantes médicinales. La prise de ces plantes durant les repas aiderait à un meilleur contrôle post-prandial de la glycémie, particulièrement chez les personnes à risque.

Évaluation de l’effet antidiabétique de plantes médicinales de la forêt boréale et identification des principes actifs de deux espèces prometteuses

Le diabète de type 2 est une maladie chronique dont l’incidence est en augmentation continuelle. Le risque de développer le diabète de type 2 chez les populations autochtones du Canada est de trois à cinq fois plus élevé que le reste de la population canadienne. La forêt boréale comporte plusieurs plantes médicinales ayant un potentiel pour le traitement ou la prévention du diabète. Certaines de ces plantes font partie de la médecine traditionnelle et alternative Crie. Des enquêtes ethnobotaniques ont amené notre équipe de recherche à identifier 17 extraits de plantes médicinales utilisées par les Cris d’Eeyou Istchee (Baie James, Québec) pour traiter les symptômes du diabète. Parmi ces extraits, certains ont montré des activités anti-diabétiques au niveau des cellules musculaires, des adipocytes et dans des études in vivo réalisées chez des animaux. Le but de cette thèse est d’élucider l’effet de ces 17 plantes sur l’homéostasie hépatique de glucose, d’identifier l’espèce la plus prometteuse et isoler ces constituants actifs. De même, le bleuet nain du genre Vaccinium angustifolium fait partie de la forêt boréale canadienne et est connu pour ses activités anti-diabétiques. Une biotransformation du jus de bleuet lui confère une activité antioxydante accrue et un profil biologique différent. Le deuxième but de cette thèse est d’élucider les mécanismes d’action par lesquels le jus de bleuet biotransformé (BJ) exerce son effet anti-diabétique et d’identifier ses principes actifs. Les résultats ont montré que trois extraits de plantes Cris se sont démarqués par leur effet sur l’homéostasie hépatique de glucose. Picea glauca exerce son effet en diminuant la production de glucose alors que Larix laricina agit en augmentant le stockage de glucose. Abies balsamea a montré le profil le plus prometteur, elle agit simultanément en diminuant l’activité de la Glucose-6-phosphatase (G6Pase) via la stimulation des voies insulino-dépendante et - indépendante et en augmentant l’activité de la Glycogène synthétase (GS) suite à la phosphorylation de la Glycogène synthase kinase-3. Le fractionnement de l’extrait d’Abies balsamea guidé par les deux bioessais a mené à l’isolation de trois composés actifs; l’acide abiétique (AA), l’acide déhydroabiétique (DAA) et le squalène (SQ). Les principes actifs ont montré le même mécanisme d’action que l’extrait brut en diminuant l’activité de la G6Pase et augmentant celle de la GS ainsi qu’en activant les voies de signalisation impliquées. Le DAA ii s’est démarqué par son effet le plus puissant et très comparable à celui de l’extrait d’Abies balsamea dans toutes les expériences. De son côté le BJ a montré un effet sur la diminution de la production hépatique de glucose, l’augmentation de son stockage ainsi que l’augmentation de son transport dans le muscle. Son fractionnement guidé par les bioessais a permis d’isoler sept fractions dont trois étaient les plus actives. L’identification des constituants de ces fractions actives a mené à isoler quatres composés phénoliques; l’acide chlorogénique, l’acide gallique, l’acide protocatéchique et le catéchol. Le catéchol s’est démarqué avec ses effets les plus puissants en diminuant l’activité de la G6Pase, augmentant celle de la GS et en stimulant le transport de glucose dans le muscle. Les résultats de cette thèse indiquent que la diminution de la production hépatique de glucose peut s’ajouter au profil anti-diabétique de certaines plantes médicinales Cries et surtout à celui d’A.balsamea dont les composés actifs peuvent aider dans le développement de nouvelles molécules anti-diabétiques. De plus, les résultats de cette thèse ont montré que l’activité antidiabétique du BJ implique le contrôle de l’homéostasie de glucose au niveau du foie et du muscle. L’identification du catéchol comme principe actif avec potentiel anti-diabétique prometteur pourra servir pour des fins thérapeutiques ultérieures.

Effect of leaf extract of Aegle marmelose in diabetic rats

Alloxan induced animal model was used to evaluate the potential antidiabetic effect of A . marmelose leaf extract. The diabetic animals were given insulin injection and another group A . marmelose leaf extract orally. It maintained the weight of the animals near to the control rats but a significant decrease in weight was noted it diabetic animals without any treatment . The blood elucose level in 'treated animals were near to that of control ones. Also a significantly increased glucose tolerance was observed in animals orally given the leaf extract prior to the experiment . A significant decrease in liver glycogen ( 1.24 -.07 s!/ 100 g of wet tissue) was observed in diabetic rats which was brought to almost the normal level (I.84- . 14 g; 100 g) with Icaf extract treatment. Blood urea and serum cholesterol increased ( 62.66 - 3.50 and 192.67± 13.64 mg' dl) significantly in alloxxan diabetic rats . The leaf extract treatment decreased the blood urea and serum . cholesterol (37.83 - 3.97 and 99?0±8.43 mg/dl ) to that of control ones . A similar effect was seen with insulin treatment . The results indicate that the active principle in .4. marmelose leaf extract has similar hypoglycaemic activity to insulin treatment.

“Effect of spice oleoresins in microbial decontamination and their potential application in quality stabilization of Tuna (Euthynnus affinis) during storage”

Unprocessed seafood harbor high number of bacteria, hence are more prone to spoilage. In this circumstance, the use of spice in fish for reduction of microorganism can play an important role in seafood processing. Many essential oils from herbs and spices are used widely in the food, health and personal care industries and are classified as GRAS (Generally regarded as safe) substances or are permitted food additives. A large number of these compounds have been the subject of extensive toxicological scrutiny. However, their principal function is to impart desirable flavours and aromas and not necessarily to act as antimicrobial agents. Given the high flavour and aroma impact to plant essential oils, the future for using these compound as food preservatives lies in the careful selection and evaluation of their efficacy at low concentrations but in combination with other chemical preservatives or preservation processes. For this reason they are worth of study alone or in combination with processing methods in order to establish if they could extend the shelf-life of foods. In this study, the effect of the spices, clove, turmeric, cardamom, oregano, rosemary and garlic in controlling the spoilage and pathogenic bacteria is investigated. Their effect on biogenic amine formation in tuna especially, histamine, as a result of bacterial control is also studied in detail. The contribution of spice oleoresin in the sensory and textural parameters is investigated using textural profile analysis and sensory panel. Finally, the potential of spices in quality stabilization and in increasing the shelf–life of tuna during frozen storage is analysed

Development and Integration of Biocontrol Products in Branched Broomrape (Phelipanche ramosa) management in Tomato

Parasitic weeds of the genera Striga, Orobanche, and Phelipanche pose a severe problem for agriculture because they are difficult to control and are highly destructive to several crops. The present work was carried out during the period October, 2009 to February, 2012 to evaluate the potential of arbuscular mycorrhizal fungi (AMF) to suppress P. ramosa on tomatoes and to investigate the effects of air-dried powder and aqueous extracts from Euphorbia hirta on germination and haustorium initiation in Phelipanche ramosa. The work was divided into three parts: a survey of the indigenous mycorrhizal flora in Sudan, second, laboratory and greenhouse experiments (conducted in Germany and Sudan) to construct a base for the third part, which was a field trial in Sudan. A survey was performed in 2009 in the White Nile state, Sudan to assess AMF spore densities and root colonization in nine fields planted with 13 different important agricultural crops. In addition, an attempt was made to study the relationship between soil physico-chemical properties and AMF spore density, colonization rate, species richness and other diversity indices. The mean percentage of AMF colonization was 34%, ranging from 19-50%. The spore densities (expressed as per 100 g dry soil) retrieved from the rhizosphere of different crops were relatively high, varying from 344 to 1222 with a mean of 798. There was no correlation between spore densities in soil and root colonization percentage. A total of 45 morphologically classifiable species representing ten genera of AMF were detected with no correlation between the number of species found in a soil sample and the spore density. The most abundant genus was Glomus (20 species). The AMF diversity expressed by the Shannon–Weaver index was highest in sorghum (H\= 2.27) and Jews mallow (H\= 2.13) and lowest in alfalfa (H\= 1.4). With respect to crop species, the genera Glomus and Entrophospora were encountered in almost all crops, except for Entrophospora in alfalfa. Kuklospora was found only in sugarcane and sorghum. The genus Ambispora was recovered only in mint and okra, while mint and onion were the only species on which no Acaulospora was found. The hierarchical cluster analysis based on the similarity among AMF communities with respect to crop species overall showed that species compositions were relatively similar with the highest dissimilarity of about 25% separating three of the mango samples and the four sorghum samples from all other samples. Laboratory experiments studied the influence of root and stem exudates of three tomato varieties infected by three different Glomus species on germination of P. ramosa. Root exudates were collected 21or 42 days after transplanting (DAT) and stem exudates 42 DAT and tested for their effects on germination of P. ramosa seeds in vitro. The tomato varieties studied did not have an effect on either mycorrhizal colonization or Phelipanche germination. Germination in response to exudates from 42 day old mycorrhizal plants was significantly reduced in comparison to non-mycorrhizal controls. Germination of P. ramosa in response to root exudates from 21 day old plants was consistently higher than for 42 day-old plants (F=121.6; P<.0001). Stem diffusates from non-mycorrhizal plants invariably elicited higher germination than diffusates from the corresponding mycorrhizal ones and differences were mostly statistically significant. A series of laboratory experiments was undertaken to investigate the effects of aqueous extracts from Euphorbia hirta on germination, radicle elongation, and haustorium initiation in P. ramosa. P. ramosa seeds conditioned in water and subsequently treated with diluted E. hirta extract (10-25% v/v) displayed considerable germination (47-62%). Increasing extract concentration to 50% or more reduced germination in response to the synthetic germination stimulants GR24 and Nijmegen-1 in a concentration dependent manner. P. ramosa germlings treated with diluted Euphorbia extract (10-75 % v/v) displayed haustorium initiation comparable to 2, 5-Dimethoxy-p-benzoquinon (DMBQ) at 20 µM. Euphorbia extract applied during conditioning reduced haustorium initiation in a concentration dependent manner. E. hirta extract or air-dried powder, applied to soil, induced considerable P. ramosa germination. Pot experiments were undertaken in a glasshouse at the University of Kassel, Germany, to investigate the effects of P. ramosa seed bank on tomato growth parameters. Different Phelipanche seed banks were established by mixing the parasite seeds (0 - 32 mg) with the potting medium in each pot. P. ramosa reduced all tomato growth parameters measured and the reduction progressively increased with seed bank. Root and total dry matter accumulation per tomato plant were most affected. P. ramosa emergence, number of tubercles, and tubercle dry weight increased with the seed bank and were, invariably, maximal with the highest seed bank. Another objective was to determine if different AM fungi differ in their effects on the colonization of tomatoes with P. ramosa and the performance of P. ramosa after colonization. Three AMF species viz. GIomus intraradices, Glomus mosseae and Glomus Sprint® were used in this study. For the infection, P. ramosa seeds (8 mg) were mixed with the top 5 cm soil in each pot. No mycorrhizal colonization was detected in un-inoculated control plants. P. ramosa infested, mycorrhiza inoculated tomato plants had significantly lower AMF colonization compared to plants not infested with P. ramosa. Inoculation with G. intraradices, G. mosseae and Glomus Sprint® reduced the number of emerged P. ramosa plants by 29.3, 45.3 and 62.7% and the number of tubercles by 22.2, 42 and 56.8%, respectively. Mycorrhizal root colonization was positively correlated with number of branches and total dry matter of tomatoes. Field experiments on tomato undertaken in 2010/12 were only partially successful because of insect infestations which resulted in the complete destruction of the second run of the experiment. The effects of the inoculation with AMF, the addition of 10 t ha-1 filter mud (FM), an organic residues from sugar processing and 36 or 72 kg N ha-1 on the infestation of tomatoes with P. ramosa were assessed. In un-inoculated control plants, AMF colonization ranged between 13.4 to 22.1% with no significant differences among FM and N treatments. Adding AMF or FM resulted in a significant increase of branching in the tomato plants with no additive effects. Dry weights were slightly increased through FM application when no N was applied and significantly at 36 kg N ha-1. There was no effect of FM on the time until the first Phelipanche emerged while AMF and N application interacted. Especially AMF inoculation resulted in a tendency to delayed P. ramosa emergence. The marketable yield was extremely low due to the strong fruit infestation with insects mainly whitefly Bemisia tabaci and tomato leaf miner (Tuta absoluta). Tomatoes inoculated with varied mycorrhiza species displayed different response to the insect infestation, as G. intraradices significantly reduced the infestation, while G. mosseae elicited higher insect infestation. The results of the present thesis indicate that there may be a potential of developing management strategies for P. ramosa targeting the pre-attachment stage namely germination and haustorial initiation using plant extracts. However, ways of practical use need to be developed. If such treatments can be combined with AMF inoculation also needs to be investigated. Overall, it will require a systematic approach to develop management tools that are easily applicable and affordable to Sudanese farmers. It is well-known that proper agronomical practices such as the design of an optimum crop rotation in cropping systems, reduced tillage, promotion of cover crops, the introduction of multi-microbial inoculants, and maintenance of proper phosphorus levels are advantageous if the mycorrhiza protection method is exploited against Phelipanche ramosa infestation. Without the knowledge about the biology of the parasitic weeds by the farmers and basic preventive measures such as hygiene and seed quality control no control strategy will be successful, however.

Prevalence of intestinal parasites in small ruminants and their senstivity to treatments with ethnobotanical remedies in Cholistan, Pakistan

Livestock production contributes substantially to the livelihoods of poor rural farmers in Pakistan; strengthening pastoral communities plays an imperative role in the country’s thrive for poverty alleviation. Intestinal helminths constitute a major threat for pastoral livestock keepers in the whole country because chronic infestation leads to distinct losses in livestock productivity, particularly the growth of young animals. Synthetic anthelmintics have long been considered the only effective way of controlling this problem but high prices, side effects and chemical residues/toxicity problems, or development of resistance, lead to their very limited use in many pastoral systems. Additionally, poor pastoralists in remote areas of Pakistan hardly have access to appropriate anthelmintic drugs, which are also relatively expensive due to the long routes of transportation. The search for new and more sustainable ways of supporting livestock keepers in remote areas has given rise to studies of ethno-botanicals or traditional plant-based remedies to be used in livestock health care. Plant-based remedies are cheap or free of cost, environmentally safe and generally create no problem of drug resistance; they thus might substitute allopathic drugs. Furthermore, these remedies are easily available in remote areas and simple to prepare and/or administer. Cholistan desert is a quite poor region of Pakistan and the majority of its inhabitants are practicing a nomadic life. The region’s total livestock population (1.29 million heads) is almost twice that of the human population. Livestock husbandry is the primordial occupation of the communities and traditionally wealth assessment was based on the number of animals, especially goats and sheep, owned by an individual. Fortunately, about 60% of this desert region is richly endowed with highly adapted grasses, shrubs and trees. This natural flora has a rich heritage of scientifically unexplored botanical pharmacopoeia. Against this background, the present research project that was conducted under the umbrella of the International Center for Development and Decent Work at Kassel University, focused on a development aspect: in the Cholistan desert region it was firstly examined how pastoralists manage their livestock, which major health problems they face for the different animal species, and which of the naturally occurring plants they use for the treatment of animal diseases (Chapter 2). For this purpose, a baseline survey was carried out across five locations in Cholistan, using a structured questionnaire to collect data from 100 livestock farmers (LF) and 20 local healers (LH). Most of LF and LH were illiterate (66%; 70%). On average, LH had larger herds (109 animals) than LF (85 animals) and were more experienced in livestock husbandry and management. On average LF spent about 163 Euro per year on the treatment of their livestock, with a huge variability in expenditures. Eighty-six traditional remedies based on 64 plants belonging to 43 families were used. Capparaceae was the botanical family with the largest number of species used (4), followed by Chenopodiaceae, Poaceae, Solanaceae and Zygophyllaceae (3). The plants Capparis decidua (n=55 mentions), Salsola foetida (n=52), Suaeda fruticosa (n=46), Haloxylon salicornicum (n=42) and Haloxylon recurvum (n=39) were said to be most effective against the infestations with gastrointestinal parasites. Aerial parts (43%), leaves (26%), fruits (9%), seeds and seed oils (9%) were the plant parts frequently used for preparation of remedies, while flowers, roots, bulbs and pods were less frequently used (<5%). Common preparations were decoction, jaggery and ball drench; oral drug administration was very common. There was some variation in the doses used for different animal species depending on age, size and physical condition of the animal and severity of the disease. In a second step the regionally most prevalent gastrointestinal parasites of sheep and goats were determined (Chapter 3) in 500 animals per species randomly chosen from pastoral herds across the previously studied five localities. Standard parasitological techniques were applied to identify the parasites in faecal samples manually collected at the rectum. Overall helminth prevalence was 78.1% across the 1000 animals; pure nematode infestations were most prevalent (37.5%), followed by pure trematode (7.9%), pure cestode (2.6%) and pure protozoa infestations (0.8%). Mixed infestations with nematodes and trematodes occurred in 6.4% of all animals, mixed nematode-cestode infestations in 3.8%, and all three groups were found in 19.1% of the sheep and goats. In goats more males (81.1%) than females (77.0%) were infested, the opposite was found in sheep (73.6% males, 79.5% females). Parasites were especially prevalent in suckling goats (85.2%) and sheep (88.5%) and to a lesser extent in young (goats 80.6%, sheep 79.3%) and adult animals (goats 72.8%, sheep 73.8%). Haemonchus contortus, Trichuris ovis and Paramphistomum cervi were the most prevalent helminths. In a third step the in vitro anthelmintic activity of C. decidua, S. foetida, S. fruticosa, H. salicornicum and H. recurvum (Chapter 2) was investigated against adult worms of H. contortus, T. ovis and P. cervi (Chapter 3) via adult motility assay (Chapter 4). Various concentrations ranging from 7.8 to 500 mg dry matter/ml of three types of extracts of each plant, i.e. aqueous, methanol, and aqueous-methanol (30:70), were used at different time intervals to access their anthelmintic activity. Levamisol (0.55 mg/ml) and oxyclozanide (30 mg/ml) served as positive and phosphate-buffered saline as negative control. All extracts exhibited minimum and maximum activity at 2 h and 12 h after parasite exposure; the 500 mg/ml extract concentrations were most effective. Plant species (P<0.05), extract type (P<0.01), parasite species (P<0.01), extract concentration (P<0.01), time of exposure (P<0.01) and their interactions (P<0.01) had significant effects on the number of immobile/dead helminths. From the comparison of LC50 values it appeared that the aqueous extract of C. decidua was more potent against H. contortus and T. ovis, while the aqueous extract of S. foetida was effective against P. cervi. The methanol extracts of H. recurvum were most potent against all three types of parasites, and its aqueous-methanol extract was also very effective against T. ovis and P. cervi. Based on these result it is concluded that the aqueous extract of C. decidua, as well as the methanol and aqueous-methanol extract of H. recurvum have the potential to be developed into plant-based drugs for treatment against H. contortus, T. ovis and P. cervi infestations. Further studies are now needed to investigate the in vivo anthelmintic activity of these plants and plant extracts, respectively, in order to develop effective, cheap and locally available anthelmintics for pastoralists in Cholistan and neighboring desert regions. This will allow developing tangible recommendations for plant-based anthelminthic treatment of sheep and goat herds, and by this enable pastoralists to maintain healthy and productive flocks at low costs and probably even manufacture herbal drugs for marketing on a regional scale.