In vitro efficacy of selected medicinal plants from Cholistan desert, Pakistan, against gastrointestinal helminths of sheep and goats

Gastrointestinal helminths are a major constraint to small ruminants in extensive husbandry systems of tropical regions. Yet, unavailability, high prices, side effects, and development of parasite resistance often limit the use of synthetic anthelmintics. Traditional medicinal plants might be an effective low-cost alternative. Therefore the in vitro anthelmintic activity of leaf extracts of the ligneous plants Capparis decidua, Salsola foetida, Suaeda fruticosa, Haloxylon salicornicum, and Haloxylon recurvum from Cholistan, Pakistan, was investigated against adult worms of Haemonchus contortus, Trichuris ovis, and Paramphistomum cervi. Various concentrations (from 7.8 to 500 mg dry matter ml^(−1)) of three extracts (aqueous, methanol, and aqueous-methanol) of each plant were tested at different time intervals for their anthelmintic activity via adult motility assay. Plant species (p<=0.01), extract type (p<=0.001), parasite species (p<=0.001), extract concentration (p<=0.001), time of exposure (p<=0.001) and their interactions (p<=0.001) affected the number of immobile or dead helminths. The 50% lethal concentration (LC_(50)) values indicated that the methanol and aqueous-methanol extracts of C. decidua, H. recurvum, and H. salicornicum as well as the methanol extract of S. fruticosa have the potential to be developed into plant-based remedies against the studied helminths. Further studies are needed to investigate the in vivo anthelmintic activity of these extracts, in order to develop effective, cheap and locally available anthelmintics for pastoralists in Cholistan and neighbouring desert regions.

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.

Urban Grass and Grass-Leaf Litter Mixtures as Source for Bioenergy Recovery

Städtische Biomassen der Grünflächen bilden eine potentielle, bisher weitgehend ungenutzte Ressource für Bioenergie. Kommunen pflegen die Grünflächen, lassen das Material aber verrotten oder führen es Deponien oder Müllverbrennungsanlagen zu. Diese Praxis ist kostenintensiv ohne für die Verwaltungen finanziellen Ausgleich bereitzustellen. Stattdessen könnte das Material energetisch verwertet werden. Zwei mögliche Techniken, um Bioenergie zu gewinnen, wurden mit krautigem Material des städtischen Straßenbegleitgrüns untersucht i) direkte anaerobe Fermentation (4 Schnitte im Jahr) und ii) „Integrierte Festbrennstoff- und Biogasproduktion aus Biomasse“ (IFBB), die Biomasse durch Maischen und mechanisches Entwässern in einen Presssaft und einen Presskuchen trennt (2 Schnitte im Jahr). Als Referenz wurde die aktuelle Pflege ohne Verwertungsoption mitgeführt (8faches Mulchen). Zusätzlich wurde die Eignung von Gras-Laub-Mischungen im IFBB-Verfahren untersucht. Der mittlere Biomasseertrag war 3.24, 3.33 und 5.68 t Trockenmasse ha-1 jeweils für die Pflegeintensitäten Mulchen, 4-Schnitt- und 2-Schnittnutzung. Obwohl die Faserkonzentration in der Biomasse der 2-Schnittnutzung höher war als im Material der 4-Schnittnutzung, unterschieden sich die Methanausbeuten nicht signifikant. Der Presskuchen aus dem krautigen Material des Straßenbegleitgrüns hatte einen Heizwert von 16 MJ kg-1 Trockenmasse, während der Heizwert des Presskuchens der Gras-Laub-Mischung in Abhängigkeit vom Aschegehalt zwischen 15 und 17 MJ kg-1 Trockenmasse lag. Der Aschegehalt der Mischungen war höher als der Grenzwert nach DIN EN 14961-6:2012 (für nicht-holzige Brennstoffe), was auf erhöhte Bodenanhaftung auf Grund der Erntemethoden zurückzuführen sein könnte. Der Aschegehalt des krautigen Materials vom Straßenrand hielt die Norm jedoch ein. Die Elementkonzentration (Ca, Cl, K, Mg, N, Na, P, S, Al, Cd, Cr, Cu, Mn, Pb, Si, Zn) im krautigen Material war generell ähnlich zu Landwirtschafts- oder Naturschutzgrünland. In den Mischungen nahm die Elementkonzentration (Al, Cl, K, N, Na, P, S, Si) mit zunehmendem Laubanteil ab. Die Konzentration von Ca, Mg und der Neutral-Detergenz-Fasern stieg hingegen an. Die IFBB-Technik reduzierte die Konzentrationen der in der Verbrennung besonders schädlichen Elemente Cl, K und N zuverlässig. Außer den potentiell hohen Aschegehalten, wurde während der Untersuchungen kein technischer Grund entdeckt, der einer energetischen Verwertung des getesteten urbanen Materials entgegenstehen würde. Ökonomische, soziale und ökologische Auswirkungen einer Umsetzung müssen beachtet werden. Eine oberflächliche Betrachtung auf Basis des bisherigen Wissens lässt hoffen, dass eine bioenergetische Verwertung städtischen Materials auf allen Ebenen nachhaltig sein könnte.