Introduction

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

Aim of the trial

The aim of this trial was to evaluate the microbicidal effect of cimenol ring against Clostridium perfringens (CP) by observing the reduction of CFU in culture plates.

Methodology

1. Prepare the following dilutions of cimenol ring: 1:10000, 1:5000 and 1:2500, corresponding to 100, 200 and 400 ppm, respectively
2. Add these dilutions to porcine enterocyte cultures (IPEC-J2 cells)
3. Add CP to the cultures and incubate them
4. Aspirate the culture medium and resuspend the cellular fraction
5. Perform serial dilutions with the suspensions
6. Deep seed the dilutions in plates with agar TSC and incubate them
7. Plates:

• Negative control: without CP and without cimenol
• Positive control: without CP, without cimenol
• AN 1:10000: lysed with CP and cimenol at 100 ppm
• AN 1:5000: lysed with CP and cimenol at 200 ppm
• AN 1:2500: lysed with CP and cimenol at 400 ppm

Observe CP CFU (CFU/plate)

Results

Images show CP growth in the different plates.

Interpretation

• There is no CP growth in the negative control plate.
• CP growth is greater in the positive control (CP without treatment).
• CP growth is lower with cimenol ring. There is a dose-effect relation (at higher doses of cimenol, smaller CP growth).
• The plate corresponding to cimenol at 400 ppm only has 4 CP colonies. This means that CP growth is almost completely inhibited.

Conclusions

Cimenol ring has a biocidal effect on Clostridium perfringens, which is observed by a reduction in the CFU of intracellular bacteria in enterocytes in contact with the product.

This decrease is particularly significant when the product is administered at 400 ppm.

In conclusion, the cimenol ring can be used as an intestinal biocide to prevent Clostridium infections.

The cimenol ring is marketed under the name Alquermold Natural by Biovet, S.A.