Highly Diluted Glyphosate Mitigates Its Effects on Artemia salina: Physicochemical Implications

Abstract

Glyphosate is an herbicide widely used in agriculture but can present chronic toxicity in low concentrations. Artemia salina is a common bio-indicator of ecotoxicity; it was used herein as a model to evaluate the effect of highly diluted-succussed glyphosate (potentized glyphosate) in glyphosate-based herbicide (GBH) exposed living systems. Artemia salina cysts were kept in artificial seawater with 0.02% glyphosate (corresponding to 10% lethal concentration or LC10) under constant oxygenation, luminosity, and controlled temperature, to promote hatching in 48 h. Cysts were treated with 1% (v/v) potentized glyphosate in different dilution levels (Gly 6 cH, 30 cH, 200 cH) prepared the day before according to homeopathic techniques, using GBH from the same batch. Controls were unchallenged cysts, and cysts treated with succussed water or potentized vehicle. After 48 h, the number of born nauplii per 100 µL, nauplii vitality, and morphology were evaluated. The remaining seawater was used for physicochemical analyses using solvatochromic dyes. In a second set of experiments, Gly 6 cH treated cysts were observed under different degrees of salinity (50 to 100% seawater) and GBH concentrations (zero to LC 50); hatching and nauplii activity were recorded and analyzed using the ImageJ 1.52, plug-in Trackmate. The treatments were performed blind, and the codes were revealed after statistical analysis. Gly 6 cH increased nauplii vitality (p = 0.01) and improved the healthy/defective nauplii ratio (p = 0.005) but delayed hatching (p = 0.02). Overall, these results suggest Gly 6cH treatment promotes the emergence of the more GBH-resistant phenotype in the nauplii population. Also, Gly 6cH delays hatching, another useful survival mechanism in the presence of stress. Hatching arrest was most marked in 80% seawater when exposed to glyphosate at LC10. Water samples treated with Gly 6 cH showed specific interactions with solvatochromic dyes, mainly Coumarin 7, such that it appears to be a potential physicochemical marker for Gly 6 cH. In short, Gly 6 cH treatment appears to protect the Artemia salina population exposed to GBH at low concentrations.

Environmental Homeopathy: Homeopathic Potencies Regulate the Growth and Toxicity of Raphidiopsis raciborskii (cyanobacteria) and Can be Tracked Physico-Chemically. Part 2: Physico-chemical Results

“…An essential detail of the method used is how samples are prepared before interacting with the dyes, and this has been described and standardized in previous studies. 18,19,22,26 For all samples, regardless of origin (potentized medicines, water, culture media), a further potency (1cH) of each sample is made in sterile pure water or 30% hydroalcoholic solution (according to the protocol), previously filtered in a 0.22-micrometer mesh filter to avoid eventual contamination. Therefore the sample added to dye solutions corresponds to 1% of the original content.…”

“…12 These physico-chemical changes can be monitored through interactions with solvatochromic dyes. [18][19][20][21][22][23][24][25] A recent systematic experimental study has also observed correspondence between biological effects and physico-chemical changes. 26 Solvatochromic dyes have been found to be probes able to track homeopathic potencies both in the laboratory [18][19][20][21][23][24][25][26] and in field studies in the natural environment.…”

“…[18][19][20][21][22][23][24][25] A recent systematic experimental study has also observed correspondence between biological effects and physico-chemical changes. 26 Solvatochromic dyes have been found to be probes able to track homeopathic potencies both in the laboratory [18][19][20][21][23][24][25][26] and in field studies in the natural environment. 22 The nature of the interaction between solvatochromic dyes and homeopathic potencies is yet to be elucidated, but the evidence so far indicates that the unique electronic structure and behavior of these dyes is crucial to the interaction and that the interaction involves an increase in their polarity, which can be measured spectroscopically in the UV-visible region of the electromagnetic spectrum.…”

“…[20][21][22][23][24][25] Different solvatochromic dyes respond to homeopathic potencies according to their structure, so selective detection and identification of various homeopathic potencies may be possible. 18,19,26 The present study aims, among others, to see if this possibility can be realized.…”

“…Indeed, this effect can be reflected in the absorbance measurement using a spectrophotometer, as seen previously. [18][19][20][21][22][23][24][25][26] In addition, a static magnetic field, being strong enough, can a priori provide a more stable magnetic interaction between dye and solvent molecules 24 through the alignment of the dipoles so that sharper differences between homeopathic potencies and controls may be identified. This fact would imply an increase in the method's sensitivity.…”