Effectiveness of ultra high diluted arsenic is a function of succussion number as evidenced by wheat germination test and droplet evaporation method

  • Lucietta Betti
  • Grazia Trebbi
  • Maria Olga Kokornaczyk
  • Daniele Nani
  • Maurizio Peruzzi
  • Maurizio Brizzi
Keywords: Succussion number, ultra high dilutions, arsenic, wheat germination, droplet evaporation method

Abstract

Background: In the preparation of ultra high dilutions (UHD) each dilution step is followed by a succussion. Whereas the physical basis of succussion have been well studied [1], there are only few papers regarding the effects of succussion number (NS) [2]. In previous studies [3, 4] we showed that arsenic at UHD applied on arsenic stressed wheat seeds stimulates the germination rate. Aims: Investigate whether NS applied between the dilution-steps influences the UHD effectiveness. Methodology: Plant material and stress treatment: Wheat seeds (Triticum aestivum L.) of cv. Pandas were stressed by 30min of poisoning with 5mM As2O3 aqueous solution [3, 4]. Classes of treatment: Undiluted and unsuccussed H2O (C), H2O and As2O3 both at the 45th decimal dilution/succussion (W45x and As45x, respectively). The succussion was handmade and NS differed for each treatment (NS= 4, 8, 16, 32, 40, 70, 100). Biological model: In each Petri dish 36 seeds were placed and watered with 20ml of treatment. After 96 hours non-germinated seeds were counted. The experiment was repeated twice and each one consisted in 6 Petri dishes per treatment and 6 trials. The results were analyzed by Poisson test [3]. Droplet evaporation method: The droplet patterns [5] were prepared out of the stressed wheat samples treated either with As45x, W45x (NS=8, 32, 70) or C. The experiment was performed in a 3 day repetition, 3 replicates per day, 5 droplets per replicate. The patterns were evaluated by means of the ImageJ software for their local connected fractal dimension (LCFD) values. The data was analyzed by means of the analysis of variance. Multiple mean comparison was carried out by Turkey’s HSD test. Results: NS strongly influenced both the biological effectiveness of the treatment (i) and the LCFD of the DEM patterns (ii). In particular: (i) the rate of germinated seeds significantly increased vs. C following treatments with NS≥32 for both As45x and W45x; (ii) all As45x (NS=8, 32, 70) and W45x with NS=70 significantly increased the LCFD of DEM patterns vs. C. Both approaches showed that UHD effects increase with the growing NS, even if there was a non complete correspondence of the significance levels. Conclusions: Our results show that NS is an important parameter influencing the UHD effectiveness.
Published
2013-08-26
Section
Plant and Microorganism Research