A controlled in vitro study of the antimicrobial effectiveness of Colibacillinum against E. coli

Abstract

Background Due to the rise of antibiotic resistant bacterial infections, alternative methods of treatment need to be explored. Homoeopathic medicine is based on the principle of ‘like cures like’ (O’Reilly, 1996) i.e. the symptoms the substance can cause, it can cure which is the underlying rationale for this study. Colibacillinum is a remedy prepared from an enteropathogenic strain of E. coli, this remedy is already used in clinical practice against chronic cystitis (Leupen, 2010) caused by E. coli, however, an extensive literature search has revealed there to be no empirical investigation into its antibiotic capacity. This study aimed to show whether or not this remedy demonstrates antimicrobial capacity against E. coli in an in vitro setting. Parallell homoeopathic potencies of this remedy and one prepared from a uropathogenic strain of E. coli were tested for antimicrobial effects against enteropathogenic and uropathogenic strains of E. coli in vitro using the disc diffusion method. Aim of the study The aim of this controlled in vitro study is to determine the antimicrobial effectiveness of parallel potencies of the homoeopathic remedy Colibacillinum (manufactured from a uropathogenic strain and enteropathogenic strains respectively) against uropathogenic and enteropathogenic cultures of E. coli in vitro by means of the disc diffusion assay method. Methodology Measurements were by means of the disc diffusion essay. For this experiment thirty Mueller-Hinton plates were prepared and inoculated with each test bacteria in turn. Fifteen plates were inoculated with Uropathogenic strain of E. coli and the remaining 15 plates were inoculated with Uropathogenic strain of E. coli. A sterile 5mm Whatman® filter paper number 4 discs were individually inoculated with test substances 3CH,9CH,30CH and 200CH potencies and the controls, negative (43% ethanol) and positive control (Ciprofloxacin) using a micropipette, before being allowed to dry in the incubator. A Ciprofloxacin antibiotic (positive control) was included in the experiment with sole purpose of accounting for plate-plate variations in the pharmacological sensitivity of the same specie of bacteria. The plates were incubated at 37°C, and the zones of inhibition measured with a pair of Vernier calipers at 24 hours. Data entry was done using the SPSS statistical package. ANOVA was used to compare the differences between the test and control groups, Mauchly’s Test of Sphericity for Uropathogenic prepared strain, Mauchly’s Test of Sphericity for Enteropathogenic prepared strain, Normality test.

Results The results obtained from this study showed that the Homoeopathic remedy Colibacillinum prepared from both Uropathogenic and Enteropathogenic strains displayed inhibitory effects against Enteropathogenic and Uropathogenic strains of E. coli, and exhibited statistically significance. The control group (ciprofloxacin) had the highest inhibitory effect (42.3±0.58mm) against Enteropathogenic and Uropathogenic E. coli, while the negative control (43% ethanol) had the lowest inhibitory effect (0.67±1.15mm). Colibacillinun 200CH prepared from a Uropathogenic strain of E-coli (Coli-b_U 200CH) displayed statistically significant antimicrobial effects against uropathogenic E.coli; such antimicrobial effects were significantly greater than 43% ethanol (negative control); the antimicrobial effect was however inferior to Ciprofloxacin (positive control). Colibacillinum 9CH prepared from Enteropathogenic strain of E-coli (Coli-b_E 9CH) also displayed statistically significant antimicrobial effects against enteropathogenic E.coli which were significantly greater than 43% ethanol (negative control) but inferior to Ciprofloxacin.

Conclusion This study concluded that Colibacillinum prepared from Uropathogenic and Enteropathogenic strains of E. coli, are effective in inhibiting the in vitro growth of E.coli when evaluated by means of disc diffusion. The study further confirmed that the biological (anti-microbial) activity of an ultra-high homoeopathic dilution (Coli-b_U 200CH) (1:10400) and in the case of Coli-b_U the findings support existing literature which suggests that the anti-microbial properties of homeopathic nosodes increase with potency; all hypotheses for this remedy were thus accepted. This trend was not noted for Coli-b_E in which the potency with the greatest anti-microbial effect was the 9CH, thus Colibacillinum prepared from Enteropathogenic strain (Coli-b_E) did not conform with hypothesies one, two and four that were proposed in chapter one. Despite this the confirmation of significant antimicrobial effects of a substance at this level of deconcentration (1:1018) is noteworthy.