Homeopathy for brine shrimp
Int J Mol Sci. 2023 May 30;24(11):9478.
Highly Diluted Glyphosate Mitigates Its Effects on Artemia salina: Physicochemical ImplicationsMirian Yaeko Dias de Oliveira Nagai (1), Suham Nowrooz Mohammad (1), Andreia Adelaide G Pinto (1), Ednar Nascimento Coimbra (2) , Giovani Bravin Peres (1), Ivana Barbosa Suffredini (1) , Maria Martha Bernardi (1), Alexander L Tournier (3), Igor Jerman (4), Steven John Cartwright (5), Leoni Villano Bonamin (1)
1 Research Center, Universidade Paulista, São Paulo 04026002, Brazil.
2 Faculty of Medicine, Universidade Federal de Alagoas, Maceió 57072900, Brazil.
3 Institute of Complementary and Integrative Medicine, University of Bern, 3012 Bern, Switzerland.
4 BION Institute, 1000 Ljubljana, Slovenia.
5 Cherwell Laboratory for Fundamental Research in Homeopathy, Oxford OX2 8NU, UK.
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.
Free full text:
https://www.mdpi.com/1422-0067/24/11/9478Excerpts:
“One of the ways to understand the toxic potential of substances is a set of tests with Artemia salina [2,3], also called brine shrimp.”
“As part of its reproduction cycle, Artemia salina produces cysts in its habitat. Metabolically, the cysts are inactive while they are dry, but after being immersed in saline water, they hydrate and change from biconcave to spherical shape. Under favorable environmental conditions, the first layer of the cyst opens and exposes the embryo, which is surrounded by a membrane attached to the cyst (umbrella stage). At this stage, the nauplii complete their development and emerge as free swimmers [4]. Artemia salina is sensitive to several chemicals, such as phenolic compounds, nitrite, mercury [5], and organophosphate [6]. In a stressful environment, the cyst hatching is arrested, and the embryo remains in a quiescent state called diapause [7].”
“Glyphosate [N-(phosphonomethyl) glycine] is a non-specific organophosphate that inhibits the enzyme 5-enolpyruvylshikimate-3-phosphate synthase, promoting the interruption of the essential aromatic amino acid syntheses in plants [10]. It is one of the most common herbicides used worldwide, traces of which can be found widespread in many different species [11,12,13], raising concerns about its possible action as an endocrine disruptor when present in commercial preparations at low concentrations [13,14,15].”
“The use of potentized high dilutions (serial highly diluted substances submitted to vigorous agitation) in aquatic animals has been reported in clinical and experimental studies over the last 20 years; a recent review of the field was published recently [17]. However, the use of potentized preparations in assisting living systems in recovering from exposure to chemical agents is still a matter of speculation.”
“This approach opens new possibilities for the rational use of potentized high dilutions as a valuable tool for the mitigation of the effects of environmental toxins. In addition, results from the use of solvatochromic dyes may provide support for the quantum electrodynamic (QED) based hypothesis of water electric dipole aggregates being a key element for the mechanisms of action of potentized high dilutions [24].”
“The results show protective effects of Gly 6 cH [a 100-fold dilution done 6 times, or 10^12 fold] after hatching, with an increase in vitality and an improved healthy/defective nauplii ratio. The hatching rate was also lower, with retention of nauplii development in the umbrella stage. Such a reduction in the hatching rate suggests this is a survival mechanism of nauplii in non-ideal environments. The result in this study mirrors the results previously described in nauplii exposed to mercuric chloride LC10 and treated with homeopathic potencies of mercuric chloride in a similar experimental model [8].”
“The salinity level seems significant concerning the degree of effectiveness of Gly 6 cH since the biological effects were more evident at 80% seawater salinity. It was also seen in the interaction between water samples and solvatochromic dyes, with a maximum concentration-dependent effect at 80% salinity. The presence of ionic species in solution has been observed to affect NMR relaxation values associated with homeopathic potencies in a previous study [27]. As yet, no satisfactory explanation for this phenomenon has been established other than a suggested mechanism [28].”
“The way of action of Gly 6 cH on nauplii development seems to be related to mechanisms of environmental adaptation, such as the gene expression of bio-resilience markers, as described in other aquatic species [32]. Nevertheless, the final glyphosate concentration in Gly 6 cH is too low (5.9 × 10-13 M) to produce classical biochemical effects when diluted 1:10 and 1:100 in water (reaching about 6 × 10-14 M and 6 × 10-15 M), indicating the necessity to consider alternative hypotheses about its possible mechanism of action. In parallel, the influence of the salinity on the Gly 6 cH effects and the corresponding interaction between treated water and a specific solvatochromic dye (Coumarin 7) are two points suggesting that the physicochemical properties of highly diluted substances might be relevant aspects.
“Whilst the mechanism of action of potentized high dilutions is still unknown, several hypotheses have been proposed in the last decade. These include hypotheses based on pharmacologically active nanoparticles, nanobubbles, the exclusion zone of water, and so-called 'nano associates' [27,33,34,35].
“In particular, the hypothesis of coherence domains proposed by Del Giudice and collaborators in 1988 [36] could be considered one of the most plausible explanations. Following this hypothesis, it is assumed there are specifically ordered molecular aggregates that make up the mesoscopic phase of water [37,38,39]. This hypothesis is based on a quantum electrodynamic (QED) model, which predicts that quantum electrodynamic effects could lead to the formation of local resonance structures in water [36], which would be reinforced by the presence of ions [37]. This QED model could serve as a theoretical background for understanding the imprinting of molecular information from a starting solution (such as a glyphosate solution) in a polar liquid medium such as water via coherent domains of different types [40,41].
“Hence the dipole moment of water might be implicated in the formation of various types of coherent domains, depending on the solute present [24]. The formation of these structures would lock in electromagnetic frequencies coming from its environment, thereby providing a sort of fingerprint (specific memory) of the original substance [42,43,44,45,46,47]. The prevalence of a specific coherent domain pattern could be seen as a fingerprint of Gly 6 cH, for instance, and its presence, therefore, could be indirectly indicated by its specific interaction with the solvatochromic dye Coumarin 7.
“Some experimental evidence supports this theory. For example, solvatochromic dyes have been shown to act as probes that track the presence of homeopathic potencies [23,46]. Assays using solvatochromic dyes are well established, and several publications are available describing the evolution of the technique [47,48,49,50,51]. Solvatochromic dyes become more polarized in the presence of homeopathic potencies, and this appears to be a dye and potency-specific phenomenon, with some dyes responding to a particular potency and others not. Potencies may therefore have particular molecular fingerprints. The model described above is consistent with the biological particularities of high dilution effects. However, more work needs to be undertaken to confirm this hypothesis.
“The use of high dilutions to mitigate chronic or acute intoxication is not new as a therapeutic resource. References to this experimental approach are available in the literature [52,53,54,55,56].”
“Although the exact mechanism of action of homeopathic high dilutions is still unknown, it may involve electromagnetic fields or polar/charged imprints in water (or other polar solvents). This possibility, along with a previous study that seemed to show that dissolved salts affect the NMR relaxation values associated with homeopathic potencies [27], prompted an examination of the effect of varying the level of salinity (ionic strength) in solutions in which Artemia salina was challenged with GBH along with potencies thereof. The fact that mitigation of the effects of GBH by potentized highly diluted GBH was dependent on the level of salinity in solution may be important, not only in furthering our understanding of the mechanism of action of homeopathic high dilutions but also in designing future field studies in aquatic environments.”
“Conclusions
“Gly 6 cH treatment appears to produce protective effects on the development of Artemia salina exposed to GBH at low concentrations. These effects can be summarized as (a) an increase in nauplii vitality, (b) the selection of healthier nauplii during hatching, (c) an optimal protective effect in a specific range of water salinity, (d) the protective effect of Gly 6 cH can be seen as an increase in bio-resilience process (e) Coumarin 7 was identified as a potential physicochemical marker for Gly 6 cH activity in real-life conditions, (f) the theoretical concept of quantum coherence domains gives a plausible explanation to these findings.”
