Droplet evaporation patterns of wheat seed leakages differ following treatments with arsenic at ultra-high dilution level

Introduction “Agro-homeopathy” plays an important role in sustainable agriculture providing cost-saving and residue-free plant treatments for the improvement of yield and the management of diseases and pests [1]. However, one of the main difficulties in “agro-homeopathy” is the exact choice of the right remedy and dilution, which requires the performance of many time-consuming trials. The droplet evaporation method could constitute a screening method for evaluating the efficacy of extremely low doses and high dilutions on plants. Droplet patterns prepared out of wheat seed leakages show differences in their complexity in relation to the seed vigour [2]. Here we show that this structure-forming capacity of seed leakages prepared in water or in ultra-high diluted (UHD) solutions may give a reliable picture of the treatment’s stimulating or inhibiting influence on plant vitality. Materials and Methods Part of the seeds was stressed with 0.1% As2O3 (As) aqueous solution [3]. The droplet patterns were prepared out of four samples: non-stressed (ns) seeds in water, ns-seeds in As 45x, stressed (s) seeds in water, s-seeds in As 45x. The experiment was performed in a 3 day repetition, 3 replicates per day, and 5 droplets per each replicate. The resulting patterns were evaluated by means of the ImageJ software [4] for their local connected fractal dimension (LCFD) values. The plant vigour of the four samples was evaluated by means of in vitro wheat growth [5, 6]. Results The mean LCFD of patterns deriving from s-seeds was significantly lower (1.4) vs. ns-seeds (1.7). Moreover, for both ns- and s-seeds, the LCFD was significantly higher with treatment As 45x vs. water (1.8 vs. 1.7 for ns-seeds, and 1.5 vs. 1.2 for s-seeds). The results of the growth test showed a significant vigour decrease caused by As stress (shoot length 19.7mm) vs. ns-seeds (22.4mm); a significant stimulating effect following As 45x treatment was observed for both s-seeds (25.1mm) and ns-seeds (31.4). Discussion The droplet patterns seem to be a sensitive tool for visualizing the effectiveness of UHD on seeds. The experiment showed that the pattern complexity increased with the application of the treatment for both ns- and s-seeds. The growth test provided a further verification of these results. Conclusions Further confirmations are required, but even as it is now the droplet evaporation method may be considered a promising tool for “agro-homeopathic” screening tests. Authors declare that there is no conflict of interest and that this study received no funding. References Betti L, Trebbi G, Majewsky V, Scherr C, Shah-Rossi D, Jaeger T, Baumgartner S. Use of homeopathic preparations in phytopathological models and in field trials: a critical review. Homeopathy 2009; 98: 244-266. Kokornaczyk MO, Dinelli G, Marotti I, Benedettelli S, Nani D, Betti L. Self-organized crystallization patterns from evaporating droplets of Common wheat grain leakages as a potential tool for quality analysis. TheScientificWorldJ. 2011; 11: 1712-1725. Brizzi M, Elia V, Trebbi G, Nani D, Peruzzi M, Betti L. The efficacy of ultra molecular aqueous dilutions on a wheat germination model as a function of heat and aging-time. e-CAM 2011; Article ID 696298: 11p. doi: 10.1093/ecam/nep217. Collins TJ. ImmageJ for microscopy. BioTechniques. 2007; 43: 25-30. Betti L, Brizzi M, Nani D, Peruzzi M. Effect of high dilutions of Arsenicum album on wheat seedlings from seed poisoned with the same substance. Br. Homeopath. J. 1997; 86: 86-89. Brizzi M, Lazzarato L, Nani D, Borghini F, Peruzzi M, Betti L. A biostatistical insight into the As2O3 high dilution effects on the rate and variability of wheat seedling growth. Forsch. Komplementmed. 2005; 12: 277-283.