Intestinal optimizer pronutrients are active molecules from plant extracts that, when fed to animals and once inside the intestine tract, they physiologically stimulate intestinal local immunity. Thanks to the activation of the local immune system, immune cells are ready to interrupt the life cycle of coccidia and limit the infestation. Intestinal polymorphonuclears (PMNs) are immune cells with an essential role in the anti-coccidia reactions.

OBJECTIVE

To evaluate the effect of intestinal optimizer pronutrients on PMNs isolated from the intestine of a chicken through the study of interleukin expression with and without coccidia.

EXPERIMENTAL DESIGN

Cell culture: PMNs from a chicken intestine

Coccidia collection: Sporozoites (asexual stage of the coccidia’s life cyce) of Eimeria acervulina, E. maxima, E. praecox and E. tenella at a concentration of 1.75×10⁵ spor./ml, approximately.

Treatments: Four PMNs suspensions were used:

• Control: PMNs were incubated for 3 hours without any treatment nor coccidia
• T1: PMNs were incubated for 3 horas with intestinal optimizer pronutrients
• T2: PMNs were incubated for 3 hours with coccidia at a rate 10:1
• T3: PMNs were incubated for 1 hour with intestinal optimizer pronutrients and then 2 more hours with coccidia at a ratio 10:1

Evaluation of interleukin expression: PMNs’ RNA was isolated after each cellular suspension and cDNA was synthesized to evaluate interleukin expression.

Evaluated interleukin and general functions

IL-1: It promotes chemokine and cytokine secretion by fibroblasts, macrophages, and epithelial cells; it attracts other inflammatory cells, such as macrophages, heterophiles and lymphocytes, and amplifies the immune response.

IL-12: It triggers Th1 immune response (T lymphocytes), which also plays an important role in the fight against coccidiosis (adaptative immunity), NK cell (cytotoxicity) and macrophage activation. It also increases the duration of the cellular immune response.

IL-18: It induces the activation of NK cells and cytotoxic T lymphocytes; it stimulates production of gamma interferon (activates macrophages and Th1 immune response) and increases the effects of IL-12.

RESULTS

IL-1 expression, related to the immune reaction and chemotaxis, got increased in the cells with pronutrients. This increase in interleukin expression was greater in cells with pronutrients and coccidia. This proves these plant-based molecules stimulate the immune response, particularly when there is a challenge.

IL-12 expression was the lowest in the control group, followed by cells with pronutrients and cells with coccidia. Cells with pronutrients and coccidia were the ones with greater IL-12 expression, which proves that pronutrients stimulate Th1 and cellular response, essential to fight coccidiosis, particularly when there is a challenge.

Regarding IL-18 expression, a marked increase was observed in cells with pronutrients (with and without coccidia) was observed, while the increase in cell with coccidia (only) was smaller. This proves that pronutrients enhance cellular immune response (NK and cytotoxic T lymphocytes) via IL-18, too.

CONCLUSIONS

Intestinal optimizer pronutrients stimulate the local immune system thanks to, partly, stimulating IL-1, IL-12, and IL-18 interleukin expression in intestinal PMNs.

These interleukins play an essential role in the elimination of intestinal protozoa such as Eimeria, since they enhance the immune response in a way that the immune system is capable to eliminate coccidia during the first stage of their life cycle.

This increase in interleukin expression is greater when, after the stimulation by pronutrients, cells are exposed to coccidia.

Intestinal optimizer pronutrients are a technology developed and marketed by Biovet S.A. under the trademark Alquernat® Zycox, which is available in powder premix and in liquid.