IMPROVED MILBOND-TX® “IN-VITRO AFLATOXIN B1 BINDING CAPABILITY”
Milwhite, Inc. introduced Improved Milbond-TX® to the animal and feed industry as a mycotoxin binder in 1992. Since then it has proven to be an efficient and safe product that continues to gain acceptance by the industry and provide assurance that when animals are fed diets containing aflatoxin (AF), IMTX has the ability to prevent the toxic effects associated with this mycotoxin. It is vital, when selecting any mycotoxin binder which is intended to be used in the feed, to demand to see both in-vitro and in-vivo data with regards to the ability to bind a specific mycotoxin. No mycotoxin binder should ever be purchased if it has not been tested under “conditions of actual use” which is in the animal. The reason for this is because some mycotoxin binders that have been tested have the ability to bind 100% of a specific mycotoxin in the test tube, but in the animal the same binder has 0% binding capability. This does not mean that in-vitro testing is unimportant. So, one might ask: Why go to all the trouble of testing a binder in-vitro if in-vivo testing is much more important? The reason is because in-vitro testing is a very good laboratory tool to use in order to screen different mycotoxin binders.
Thus, a high ability to bind a specific mycotoxin in the test tube wouldopen the window for further testing as an effective mycotoxin binder in the animal. Also, in-vivo testing is much more expensive and time consuming than in-vitro testing and separating out the binding potential of a product by first conducting in-vitro screening saves a lot of time, effort and cost.
During its development, IMTX was tested extensively “inhouse” by Milwhite, Inc. in their laboratory. When convinced it had promise as a binder, Dr. David Ledoux at the University of Missouri (USA) was contacted to further test IMTX because his laboratory has the ability to test a mycotoxin binder in-vitro and in-vivo. Also, being tested independently in well-designed and controlled experiments would provide consumers with more confidence and assurance that IMTX is a safe and effective mycotoxin binder. The information in this issue of Milwhite’s Journal briefly summarizes the in-vitro research that Dr. Ledoux conducted prior to his testing of IMTX invivo using broilers. A complete and detailed description of the materials and methods used in this in-vitro testing of IMTX to bind AFB1 can be found in the publication of this research (Ledoux et al., 1999).
In his laboratory and working cooperatively with the Veterinary Medical Diagnostic Laboratory at the University of Missouri, Dr. Ledoux purchased purified Aflatoxin B1 (AFB1) from Sigma Chemical Company. The binding ability of IMTX was tested in four solutions with each solution having a different pH. This was done because in the animal’s digestive tract the pH can range from very acid to basic as material moves from the anterior to the posterior of the tract. IMTX was tested for its ability to bind AFB1 in 50 ml of a buffered solution at pH of 3, 5, 7 and 9 each of which contained 2 micrograms of AFB1 per ml. To each of these pH solutions containing AFB1, 0.5 grams of IMTX was added and stirred continuously for 30 minutes. Samples of these solutions were then tested for AFB1 using HPLC.
The ability of IMTX to bind AFB1 in each of the above mentioned solutions at pH of 3, 5, 7 and 9 was found to be 100%. Furthermore, in subsequent in-vitro testing at a pH of 3 to 9, IMTX was also shown to be able to bind 100% of the AFB1 in solutions containing 250, 500 and 1,250 micrograms of AFB1. The data collected in this laboratory experiment with IMTX provided the needed evidence showing that IMTX was, indeed, a prime candidate for further in-vivo research using the broiler as the experimental animal. The in-vivo research conducted by Dr. David Ledoux and his colleagues at the University of Missouri documenting the ability of IMTX to bind AFB1 will be the focus of the next issue of Milwhite’s Journal.
Note: A complete description of the in-vitro experiment conducted at the University of Missouri and the data collected in the experiment can be found in the referenced publication located in the footnote below. The information presented in this issue of Milwhite’s Journal was compiled by Dr. Orlando Osuna, Director of Health Science at Milwhite, Inc. and Dr. Richard Miles, Professor Emeritus, University of Florida, Gainesville, FL, USA.
Ledoux, D.R., G.E. Rottinghaus, A.J. Bermudez and M. Alonso-Debolt. 1999. Efficacy of a hydrated sodium calcium aluminosilicate to ameliorate the toxic effects of aflatoxin in broiler chicks. Poultry Science. 78:204-210.