Biological Photonic Crystals: Diatoms

The aim of this work was to produce a variety of fluorescent diatom cell wall material as a basis for spectroscopic investigations of the influence of the photonic structure on the emission of an incorporated laser dye. This goal was achieved by the method of in vivo-fluorochromation, in which the fluorescence dyes are incorporated by the diatom cells during cell wall formation. Several fluorescent dyes (mostly rhodamines) known as strong laser dyes, were tested for a possible application within this method. The results of this work show that half of the tested rhodamines can be applied for an in vivo-fluorochromation of diatom cells. For a successful incorporation into the diatom cell wall, a relatively low toxicity to diatom cells is necessary. Replacement of the carbon acid function at the carboxyphenyl ring of the rhodamine by a methyl or ethylester function showed to convert a rhodamine of relatively low toxicity to a rhodamine leading to severe lethal effects within the cells. In contrast to their carbon acid forms, which posses a net neutral charge of the molecule, rhodamine esters exhibit a net positive charge. The enhanced toxicological effects seem to be due to an increased accumulation of positive charged rhodamines within the mitochondria, an increased hydrophobicity due to the attachment of an alkyl substituent, an increased retention time of the dyes within the mitochondria and a therefore stronger negative effect on the mitochondrial membrane bound energy processes of the diatom cell. Therefore rhodamines with a positive net charge deriving from a methyl or ethylester function at the carboxy phenyl ring instead of a carbon acid substituent showed not to be suitable for long-term investigations/ biomineralization studies of diatoms. Investigations performed on diatom species of different orders showed that rhodamine 19, rhodamine B, and rhodamine 101 can presumably be successfully applied for in vivo-fluorochromation to all diatom species. The results obtained here can help to find further laser dyes for an in vivo-fluorochromation of diatom cells and therefore for the production of fluorescent nanostructural elements for a detailed optical investigation of the diatom cell wall. First optical measurements performed on in vivo-fluorochromated cell walls did not give any hints concerning the photonic structure of the diatom cell. Cell wall parts with different nanostructural elements were investigated and by comparison of the obtained fluorescence emission spectra, no special features that might derive from photonic structural effects could be observed. Results concerning the concentration dependent shifts within the emission spectra, as well as the decrease of fluorescence intensity of the stained cell wall structures with increasing dye concentration, depict that several effects occurring by interaction of the molecules within the cell wall can have an impact on the technical application of fluorescent cell walls. It can be assumed that the investigation of the photonic crystal behaviour and the possibility to achieve laser action within the diatom cell wall can be hampered by molecular interactions. The results give hints to prevent such obstacles. Comparison of the recent findings and state of the art of in vivo-fluorochromation of diatom cell wall material, make clear that the here presented results are of importance and can offer a considerable contribution to the development and establishment of new biosilification markers, for diatoms as well as for other biosilifying organisms.

Effects of Different Feeding Regimes on the Digestibility and Faecal Excretion of Nitrogen, Soluble Carbohydrates and Fibre Fractions in Water Buffaloes kept under Subtropical Conditions

In the course of the ‘Livestock Revolution’, extension and intensification of, among others, ruminant livestock production systems are current phenomena, with all their positive and negative side effects. Manure, one of the inevitable secondary products of livestock rearing, is a valuable source of plant nutrients and its skillful recycling to the soil-plant interface is essential for soil fertility, nutrient - and especially phosphorus - uses efficiency and the preservation or re-establishment of environmentally sustainable farming systems, for which organic farming systems are exemplarily. Against this background, the PhD research project presented here, which was embedded in the DFG-funded Research Training Group 1397 ‘Regulation of soil organic matter and nutrient turnover in organic agriculture ’ investigated possibilities to manipulate the diets of water buffalo (Bubalus bubalis L.) so as to produce manure of desired quality for organic vegetable production, without affecting the productivity of the animals used. Consisting of two major parts, the first study (chapter 2) tested the effects of diets differing in their ratios of carbon (C) to nitrogen (N) and of structural to non-structural carbohydrates on the quality of buffalo manure under subtropical conditions in Sohar, Sultanate of Oman. To this end, two trials were conducted with twelve water buffalo heifers each, using a full Latin Square design. One control and four tests diets were examined during three subsequent 7 day experimental periods preceded each by 21 days adaptation. Diets consisted of varying proportions of Rhodes grass hay, soybean meal, wheat bran, maize, dates, and a commercial concentrate to achieve a (1) high C/N and high NDF (neutral detergent fibre)/SC (soluble carbohydrate) ratio (HH), (2) low C/N and low NDF/SC ratio (LL); (3) high C/N and low NDF/SC ratio (HL) and (4) low C/N and high NDF/SC (LH) ratio. Effects of these diets, which were offered at 1.45 times maintenance requirements of metabolizable energy, and of individual diet characteristics, respectively, on the amount and quality of faeces excreted were determined and statistically analysed. The faeces produced from diets HH and LL were further tested in a companion PhD study (Mr. K. Siegfried) concerning their nutrient release in field experiments with radish and cabbage. The second study (chapter 3) focused on the effects of the above-described experimental diets on the rate of passage of feed particles through the gastrointestinal tract of four randomly chosen animals per treatment. To this end, an oral pulse dose of 683 mg fibre particles per kg live weight marked with Ytterbium (Yb; 14.5 mg Yb g-1 organic matter) was dosed at the start of the 7 day experimental period which followed 21 days of adaptation. During the first two days a sample for Yb determination was kept from each faecal excretion, during days 3 – 7 faecal samples were kept from the first morning and the first evening defecation only. Particle passage was modelled using a one-compartment age-dependent Gamma-2 model. In both studies individual feed intake and faecal excretion were quantified throughout the experimental periods and representative samples of feeds and faeces were subjected to proximate analysis following standard protocols. In the first study the organic matter (OM) intake and excretion of LL and LH buffaloes were significantly lower than of HH and HL animals, respectively. Digestibility of N was highest in LH (88%) and lowest in HH (74%). While NDF digestibility was also highest in LH (85%) it was lowest in LL (78%). Faecal N concentration was positively correlated (P≤0.001) with N intake, and was significantly higher in faeces excreted by LL than by HH animals. Concentrations of fibre and starch in faecal OM were positively affected by the respective dietary concentrations, with NDF being highest in HH (77%) and lowest in LL (63%). The faecal C/N ratio was positively related (P≤0.001) to NDF intake; C/N ratios were 12 and 7 for HH and LL (P≤0.001), while values for HL and LH were 11.5 and 10.6 (P>0.05). The results from the second study showed that dietary N concentration was positively affecting faecal N concentration (P≤0.001), while there was a negative correlation with the faecal concentration of NDF (P≤0.05) and the faecal ratios of NDF/N and C/N (P≤0.001). Particle passage through the mixing compartment was lower (P≤0.05) for HL (0.033 h-1) than for LL (0.043 h-1) animals, while values of 0.034 h-1 and 0.038 h-1 were obtained for groups LH and HH. At 55.4 h, total tract mean retention time was significantly (P≤0.05) lower in group LL that in all other groups where these values varied between 71 h (HH) and 79 h (HL); this was probably due to the high dietary N concentration of diet LL which was negatively correlated with time of first marker appearance in faeces (r= 0.84, P≤0.001), while the dietary C concentration was negatively correlated with particle passage through the mixing compartment (r= 0.57, P≤0.05). The results suggest that manure quality of river buffalo heifers can be considerably influenced by diet composition. Despite the reportedly high fibre digestion capacity of buffalo, digestive processes did not suppress the expression of diet characteristics in the faeces. This is important when aiming at producing a specific manure quality for fertilization purposes in (organic) crop cultivation. Although there was a strong correlation between the ingestion and the faecal excretion of nitrogen, the correlation between diet and faecal C/N ratio was weak. To impact on manure mineralization, the dietary NDF and N concentrations seem to be the key control points, but modulating effects are achieved by the inclusion of starch into the diet. Within the boundaries defined by the animals’ metabolic and (re)productive requirements for energy and nutrients, diet formulation may thus take into account the abiotically and biotically determined manure turnover processes in the soil and the nutrient requirements of the crops to which the manure is applied, so as to increase nutrient use efficiency along the continuum of the feed, the animal, the soil and the crop in (organic) farming systems.

Multi-site time-trend analysis of soil fertility management effects on crop production in sub-Saharan West Africa

Soil fertility constraints to crop production have been recognized widely as a major obstacle to food security and agro-ecosystem sustainability in sub-Saharan West Africa. As such, they have led to a multitude of research projects and policy debates on how best they should be overcome. Conclusions, based on long-term multi-site experiments, are lacking with respect to a regional assessment of phosphorus and nitrogen fertilizer effects, surface mulched crop residues, and legume rotations on total dry matter of cereals in this region. A mixed model time-trend analysis was used to investigate the effects of four nitrogen and phosphorus rates, annually applied crop residue dry matter at 500 and 2000 kg ha^-1, and cereal-legume rotation versus continuous cereal cropping on the total dry matter of cereals and legumes. The multi-factorial experiment was conducted over four years at eight locations, with annual rainfall ranging from 510 to 1300 mm, in Niger, Burkina Faso, and Togo. With the exception of phosphorus, treatment effects on legume growth were marginal. At most locations, except for typical Sudanian sites with very low base saturation and high rainfall, phosphorus effects on cereal total dry matter were much lower with rock phosphate than with soluble phosphorus, unless the rock phosphate was combined with an annual seed-placement of 4 kg ha^-1 phosphorus. Across all other treatments, nitrogen effects were negligible at 500 mm annual rainfall but at 900 mm, the highest nitrogen rate led to total dry matter increases of up to 77% and, at 1300 mm, to 183%. Mulch-induced increases in cereal total dry matter were larger with lower base saturation, reaching 45% on typical acid sandy Sahelian soils. Legume rotation effects tended to increase over time but were strongly species-dependent.

Effects of time-controlled and continuous grazing on total herbage mass and ground cover

Grazing practices in rangelands are increasingly recognized as a management tool for environmental protection in addition to livestock production. Long term continuous grazing has been largely documented to reduce pasture productivity and decline the protective layer of soil surface affecting environmental protection. Time-controlled rotational grazing (TC grazing) as an alternative to continuous grazing is considered to reduce such negative effects and provides pasture with a higher amount of vegetation securing food for animals and conserving environment. To research on how the grazing system affects herbage and above ground organic materials compared with continuous grazing, the study was conducted in a sub-tropical region of Australia from 2001 to 2006. The overall results showed that herbage mass under TC grazing increased to 140% in 2006 compared with the first records taken in 2001. The outcomes were even higher (150%) when the soil is deeper and the slope is gentle. In line with the results of herbage mass, ground cover under TC grazing achieved significant higher percentages than continuous grazing in all the years of the study. Ground cover under TC grazing increased from 54% in 2003 to 73%, 82%, and 89% in 2004, 2005, and 2006, respectively, despite the fact that after the high yielding year of 2004 herbage mass declined to around 2.5 ton ha^(−1) in 2005 and 2006. Under continuous grazing however there was no significant increase over time comparable to TC grazing neither in herbage mass nor in ground cover. The successful outcome is largely attributed to the flexible nature of the management in which grazing frequency, durations and the rest periods were efficiently controlled. Such flexibility of animal presence on pastures could result in higher water retention and soil moisture condition promoting above ground organic material.