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Introduction:  In 2020, a 26,849-ha state park in Mato Grosso do Sul state, Brazil, had 30% of its area damaged by fire. A homeopathic complex formulation was applied at strategic point locations in the park's springs or watercourses, aiming to mitigate the fire damage to the flora and fauna as quickly as possible. The duration of the homeopathic signal at each point was assessed using an established solvatochromic dye technique., Objective:  To evaluate the timing and the nature of the signal at each of nine point locations. We could thus identify the presence of any signal variations due to specified environmental features within the park., Methods:  Water samples were harvested from each intervention point at different times, filtered, frozen, and sent to the laboratory, where they were prepared to 1cH using filtered 30% ethanol. Methylene violet was chosen among six dyes since it was found in preliminary tests that it could trace the homeopathic complex used. In addition to simple sample testing, samples were submitted to a static and unidirectional magnetic field of 2400 Gauss (240 mT) for 15 minutes immediately before reading, which enhanced the method's sensitivity. One-way analysis of variance/Tukey test was used to identify dye absorbance changes following the analysis of water samples from the watercourse system. A correlation matrix and the Spearman r test were employed to evaluate any correlation between tracking and the pre-existing anthropic interventions at harvesting points. In all cases, α = 0.05., Results:  Four tracking patterns using the sample magnetization process were observed in relation to water samples and their effect on methylene violet solutions: no response (P2, P4), early transitory response (P5, P6, P8), late response (P1, P9), and constant response (P3, P7). P2 and P4, which could not be tracked, were correlated with permanent local anthropic disturbance., Conclusions:  Methylene violet was the best dye to track the homeopathic complex prepared specifically for this case. Tracking was facilitated by prior magnetic treatment of samples, but anthropic disturbances to the environment seem to interfere with it., Competing Interests: Sigo Homeopatia company donated the water samples in partnership with the governmental organization IMASUL. The researchers were not given any other resources from either organization., (Faculty of Homeopathy. This article is published by Thieme.)

Introduction:  In 2021, the area of CRAS (Centro de Reabilitação de Animais Selvagens - Wild Animal Rehabilitation Center), located in a state park in Campo Grande City, Mato Grosso do Sul, Brazil, was suffering from a tick infestation affecting wild animals that inhabit the area and humans that visited its trails. Following a formal technical-scientific cooperation agreement between IMASUL, an institute for the environment in Mato Grosso do Sul state, and SIGO Homeopatia, a formulated homeopathic complex ( Formula Parques Urbanos , FPU) was designed and prepared specifically to treat the animals. This environmental intervention used specially designed slow-release water biodegradable devices. Tracking the FPU signal in water was necessary to monitor and manage the intervention., Aims:  Our aims were (1) to evaluate, among six previously standardized solvatochromic dyes, which would serve as a marker for the homeopathic complex under study; and (2) to evaluate whether the chosen solvatochromic dye could map the propagation of the homeopathic complex activity throughout the stream system from water samples harvested at different locations over time., Method:  Water samples were harvested from each point at different times, filtered, frozen, and sent to the laboratory, where they were prepared at 1cH potency for analysis using 30% ethanol as the vehicle. Solvatochromic dyes were used to analyze the samples since they alter their absorbance when in contact with homeopathic potencies. Of the six dyes tested, Coumarin 7 was found to be the most suitable for tracking the FPU complex. A static and average unidirectional magnetic field of 2,400 Gauss (240 mT), generated by a neodymium magnet, was applied to the samples immediately before reading., Results:  There were significant differences in the delta absorbance of dyes when adding treated/potentized water samples, making it possible to map the propagation of the FPU signal throughout the park over time. The signals were identifiable at the same point 1 minute and 32 days after the insertion of the device into the water. These signals were also identifiable after 75 minutes and 8 days at a point far from the insertion place., Conclusion:  Coumarin 7 was the best marker for the homeopathic complex (FPU) used to treat the wild animals living in the park. The microplates/enzyme-linked immunosorbent assay reader method and the application of a magnetic field to samples were shown to be effective in tracing homeopathic signals by changes in dye absorbance ( p ≤ 0.02) in a real-life situation, with large volumes of water, involving many environmental variables, and over large distances., Competing Interests: The samples were made available for analysis by SIGO Homeopatia company in partnership with the governmental organization IMASUL (Instituto do Meio Ambiente do Mato Grosso do Sul). No other resource was given to the researchers., (Faculty of Homeopathy. This article is published by Thieme.)

Introduction:  Cyanobacteria are microorganisms found in many parts of the world and several genera, such as Raphidiopsis raciborskii , are producers of cyanotoxins. Homeopathic potencies have been found to modulate toxicity in different biological models, and the present study endeavors to discover whether this might also be the case with cyanobacteria., Objectives:  Our objective was to investigate the possible effects of homeopathic potencies on the resilience of Artemia franciscana (brine shrimp) embryos to saxitoxin (STX; cyanotoxin) and on controlling the growth of R. raciborskii in vitro ., Method:   A. franciscana cysts were cultivated in seawater in 96-well plates to evaluate the hatching rate and vitality, plus the gene expression of heat shock proteins (HSPs), after being challenged with R. raciborskii extract containing 2.5 µg/L of STX and treated with different homeopathic potencies. Untreated wells were used as controls ("base-line"). Potencies were chosen from a screening process based on seven selected homeopathic preparations according to the similitude of STX symptoms ( Sulphur , Zincum metallicum , Nitric acidum , Plumbum metallicum , Mercurius solubilis , Phosphoric acidum , Isotherapic from R. raciborskii extract; all at 6cH, 30cH and 200cH). Cultures of R. raciborskii maintained in an artificial seawater medium were equally treated with screened homeopathic potencies selected from the same list but specifically for their growth control as a function of time., Results:  A 15% lower rate of hatching of A. franciscana cysts was observed after treatment with Nitric acidum 6cH in comparison with baseline ( p  = 0.05). A complete toxicity reversal was seen after treatment with Isotherapic 200cH, with a 23-fold increase of Hsp 26 gene expression ( p  = 0.023) and a 24-fold increase of p26 gene expression ( p ≤ 0.001) in relation to baseline. Nitric acidum 200cH and Mercurius solubilis 30cH limited the exponential growth of cyanobacteria up to 95% and 85% respectively ( p ≤ 0.003) in relation to baseline. Succussed water presented only a transitory 50% inhibition effect., Conclusion:  Isotherapic 200cH improved A. franciscana bioresilience to STX; Nitric acidum 200cH and Mercurius solubilis 30cH showed the optimal performance on limiting R. raciborskii growth. The results point to the potential of homeopathic potencies to mitigate environmental problems related to water quality., Competing Interests: None declared., (Faculty of Homeopathy. This article is published by Thieme.)

Introduction:  The control of cyanobacterial toxicity and growth by homeopathic potencies was described in Part 1 of this two-part report. Here, a parallel approach characterized the physico-chemical features of the potencies used and the liquid media treated with them, correlating these results with their respective biological effects., Objectives:  Our objective was to establish if physico-chemical parameters can track homeopathic potencies in seawater or artificial seawater medium (ASM)-1 and to discover whether these parameters correlate with previously described biological effects., Method:   Artemia franciscana (brine shrimp) cysts were cultivated in seawater challenged with Raphidiopsis raciborskii extract and treated with different homeopathic potencies chosen from a screening process. Cultures of R. raciborskii maintained in ASM-1 were also treated with previously screened homeopathic potencies, and their growth was monitored as a function of time. The physico-chemical properties of the treated media (seawater or ASM-1) were evaluated by their interaction with solvatochromic dyes and changes in pH, conductivity and temperature., Results:  Coumarin 7 was found to be a marker for Nitric acidum 6cH and Isotherapic ( R. raciborskii extract) 200cH in seawater (analysis of variance [ANOVA], p  = 0.0015). Nile red was found to be a marker for Nitric acidum 200cH and Mercurius solubilis 30cH in ASM-1 (ANOVA, p ≤ 0.001). An increase in pH of ASM-1 and endothermic effects were observed after these treatments (two-way ANOVA, p  = 0.0001). Seawater and ASM-1 to which potencies had been added were also subjected to a constant unidirectional 2,400 Gauss static magnetic field and found to have enhanced effects on the solvatochromic dyes tested., Conclusion:  Homeopathic potencies were specifically traceable in aqueous media using solvatochromic dyes, especially when the samples were subjected to a magnetic field. Results from monitoring other physical parameters, such as pH and temperature, were less specific in relation to potency tracking. However, potency-induced endothermic effects might provide valuable thermodynamic data relating to the nature of potencies., Competing Interests: None declared., (Faculty of Homeopathy. This article is published by Thieme.)

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.

Introduction: Finding solutions to mitigate the impact of pollution on living systems is a matter of great interest. Homeopathic preparations of toxic substances have been described in the literature as attenuation factors for intoxication. Herein, an experimental study using Artemia salina and mercury chloride was developed as a model to identify aspects related to bioresilience., Aims: The aim of the study was to describe the effects of homeopathic Mercurius corrosivus (MC) on Artemia salina cysts hatching and on mercury bioavailability., Methods: Artemia salina cysts were exposed to 5.0 µg/mL of mercury chloride during the hatching phase. MC potencies (6cH, 30cH, and 200cH) were prepared in sterile purified water and poured into artificial sea water. Different controls were used (non-challenged cysts and challenged cysts treated with water, succussed water, and Ethilicum 1cH). Four series of nine experiments were performed to evaluate the percentage of cyst hatching. Soluble total mercury (THg) levels and precipitated mercury content were also evaluated. Solvatochromic dyes were used to check for eventual physicochemical markers of MC biological activity., Results: Significant delay ( p  < 0.0001) in cyst hatching was observed only after treatment with MC 30cH, compared with controls. This result was associated with an increase of THg concentration in water ( p  = 0.0018) and of chlorine/oxygen ratio ( p  < 0.0001) in suspended micraggregates, suggesting changes in mercury bioavailability. A specific interaction of MC 30cH with the solvatochromic dye ET33 ( p  = 0.0017) was found., Conclusion: Changes in hatching rate and possible changes in Hg bioavailability are postulated as protective effects of MC 30cH on Artemia salina , by improving its natural bioresilience processes., Competing Interests: None declared., (Faculty of Homeopathy. This article is published by Thieme.)

Introduction: Encephalitozoon cuniculi ( E. cuniculi ), a fungus that acts as an intracellular pathogen, causes a marked neurological syndrome in many host species and is a zoonotic concern. Although no well-established treatment for this syndrome is known, previous successful clinical experience using homeopathic phosphorus has been described in which symptom remission with no mortality occurred in 40/42 animals by means of unknown immunological mechanisms. The latter observation was the main motivation for this study., Objective: To verify, in an in-vitro model, if macrophages infected with E. cuniculi can change in function after treatment with different potencies of phosphorus., Materials and Methods: RAW 264.7 macrophages were infected with E. cuniculi in-vitro and treated with various homeopathic potencies of phosphorus. The vehicle was used as a control solution (0.06% succussed ethanol). After 1 and 24 hours, the following parameters were analyzed: parasite internalization (by the Calcofluor staining method), lysosome activity (by the acridine orange method), cytokine/chemokine production (by the MAGPIX system), and cell ultrastructure. Automatic image analysis was used when applicable, and the experiments were performed in triplicate., Results: Treatment with vehicle alone increased interleukin (IL)-6, tumor necrosis factor alpha and monocyte chemotactic protein -1 production ( p  ≤ 0.05) and reduced the number of internalized parasites ( p  ≤ 0.001). A progressive and time-dependent increase in RANTES (regulated on activation, normal T-cell expressed and secreted) and lysosome activity ( p  ≤ 0.002) was observed only after treatment with the highest potency of phosphorus ( Phos 200cH), together with decreased apoptosis rate, intense parasite digestion, and the presence of non-internalized spores., Conclusions: Phos 200 cH has a modulatory action on the activity of infected macrophages, especially a specific increase in RANTES, a key element in the prognosis of E. cuniculi -infected and of immunosuppressed patients bearing infections., Competing Interests: There is no conflict of interest related to this project., (The Faculty of Homeopathy.)