Calcium requirements are critical for layers from the pre-laying period to after the laying peak due to the high calcium demand derived from egg production. They are also still very important after week 40 of production due to skeletal calcium depletion and poor calcium absorption that takes place during that period.

Because of it, it is essential to supply a proper concentration of calcium in the diets, but also to make sure its optimal absorption in the gut.

Objective

In a study conducted during June 2016, a natural additive based on mineral conditioner pronutrients was tested in vitro to evaluate its capacity to increase calcium absorption in the gut.

Methodology

Cell culture: IPEC-J2 (porcine enterocytes.

Calcium marker: Fluo-4, a permeable derivative of the radiometric probe for calcium (Ca²⁺) to measure calcium cytosolic concentration ([Ca²⁺]c) by fluorescence. When added to the cells it crosses the cellular membrane and, once inside, cellular esterases eliminate acetoxymethyl ester groups, generating Fluo-4, which cannot cross the cellular membrane again.

Evaluation of calcium absorption: Normalized fluorescence measurements give intracellular Ca²⁺ concentration.

Treatments: for each of the following treatments (with/without pronutrients), 2 different mediums were used, one with calcium phosphate and the other without calcium (EGTA). This way, the basal concentration of intracellular calcium (EGTA cultures) could be compared with the intracellular calcium concentration after the absorption (calcium medium).

• Row 1: 12 wells with pronutrients (dilution 1:10000)
  • 6 wells with calcium phosphate 2mM
  • 6 wells with EGTA 1mm
• Row 2: 12 wells with pronutrients (dilution 1:1000)
  • 6 wells with calcium phosphate 2mM
  • 6 wells with EGTA 1mm
• Row 3: 12 wells with pronutrients (dilution 1:100)
  • 6 wells with calcium phosphate 2mM
  • 6 wells with EGTA 1mm
• Row 4: 12 wells with no treatment
  • 6 wells with calcium phosphate 2mM
  • 6 wells with EGTA 1mm
• Row 5: 12 wells with carbacol 1mM (positive control, to test that the technique is working)

Procedure: After incubation, cells were washed and incubated with Fluo-4.  Plates were incubated 1again and fluorescence was read.

Results

In those cells treated with mineral conditioner pronutrients and in presence of calcium in the medium, there is an increase in calcium intracellular concentration (greater calcium absorption).

The increase in fluorescence in treated cells was statistically significant, as showed in the picture (Row 3).

Conclusions

This trial proves that mineral conditioner pronutrients increase calcium absorption in the gut.

Thanks to it, it is easier to supply hens with the necessary calcium requirements, particularly in highly productive periods, when increasing calcium concentration in the diet is not enough.

These active principles help avoiding eggshell problems and cage fatigue syndrome.

Mineral conditioner pronutrients are marketed under the name Alquernat Magacal by Biovet S.A.