The cimenol ring is an intestinal microbicide of a botanical origin with broad bactericidal and fungicidal activities that have been proved to be useful for the prevention and treatment of necrotic enteritis caused by Clostridium perfringens (CP).

The aim of this in vitro test, conducted by a European laboratory in 2019, was to prove the bactericidal effect of an additive based on cimenol ring on CP, the etiological agent of necrotic enteritis.

Methodology

1. Prepare the following dilutions of cimenol ring: 1:10000 (0,1 kg/t), 1:5000 (0,2 kg/t) and 1:2500 (0,4 kg/t). Prepare a control dilution too (without cimenol)
2. Add the different dilutions of cimenol ring to porcine enterocyte cultures (IPEC-J2 cell line) and incubate them for 90 minutes
3. Add CP to the cultures (inoculum with a MOI inferior to 0,5) and incubate them for 18 hours
4. Evaluate with the microscope the cytopathic-cytotoxic effect caused by the pathogen

Results

Images show enterocyte cultures with CP and cimenol ring at different concentrations. Fusiform formations correspond to the enterocytes and the ones with a shiny halo to CP.

Interpretation

• There is a higher integrity of the enterocyte monolayer (spindle-shaped) and a lower presence of CP in the cimenol ring culture at 0.4 kg / t.
• A dose-effect relationship is observed: the higher the concentration, the greater the biocidal effect of the product.

Conclusions

The cimenol ring has a biocidal effect on CP, which is observed by the decrease in the presence of this bacterium in enterocyte cultures and a greater integrity of the cell monolayer when there is cimenol in the medium.

At 0.4 kg / t, the presence of CP in the culture is significantly reduced, which confirms that it is an effective concentration of cimenol ring for the control of necrotic enteritis.

Cimenol ring is marketed under the commercial name Alquermold Natural by Biovet, S.A.