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. 2023 Jul 31;13(8):1667.
doi: 10.3390/life13081667.

Box-Behnken Design for Assessing the Efficiency of Aflatoxin M1 Detoxification in Milk Using Lactobacillus rhamnosus and Saccharomyces cerevisiae

Mounir M Salem-Bekhit  1 , Omnia Karem M Riad  2 , Heba Mohammed Refat M Selim  2   3 , Sally Tohamy Kamal Tohamy  2 , Ehab I Taha  4 , Saleh A Al-Suwayeh  4 , Gamal A Shazly  4
Affiliations

Box-Behnken Design for Assessing the Efficiency of Aflatoxin M1 Detoxification in Milk Using Lactobacillus rhamnosus and Saccharomyces cerevisiae

Mounir M Salem-Bekhit et al. Life (Basel). .

Abstract

Milk contaminated with aflatoxin can lead to liver cancer. Aflatoxin B1 (AFB1), a serious animal feed contaminant, is transformed into Aflatoxin M1 (AFM1) and secreted in milk. In this study, a biological method using probiotic bacteria, Lactobacillus rhamnosus (L. rhamnosus) in combination with Saccharomyces cerevisiae (S. cerevisiae), was used to assess their antiaflatoxigenic effect in animal milk. A Box-Behnken design was used to establish the optimal ratio of L. rhamnosus and S. cerevisiae, incubation time, and temperature for efficient AFM1 detoxification from milk. To achieve this, the primary, interaction, and quadratic effects of the chosen factors were investigated. To investigate the quadratic response surfaces, a second-order polynomial model was built using a three-factor, three-level Box-Behnken design. The quantity of AFM1 was detected by the ELISA technique. The results of these experiments obtained an optimum condition in AFM1 detoxification of the three tested factors in order to maximize their effect on AFM1 detoxification in milk. The model was tested in three highly contaminated milk samples to assure the efficacy of the model. AFM1 detoxification was up to 98.4% in contaminated milk samples. These promising results provide a safe, low-cost, and low-time-consuming solution to get rid of the problem of milk contamination with AFM1.

Keywords: Box–Behnken design; L. rhamnosus; S. cerevisiae; aflatoxin M1; animal milk; antiaflatoxigenic.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The levels of AFM1 (ng/L) in ten tested raw milk samples.
Figure 2
Figure 2
Expected and observed percent AFM1 reduction values (p ≤ 0.05).
Figure 3
Figure 3
Response surface plot illustrates the effect of LR:SC ratio (X1) and incubation temperature (X2) on % AFM1 reduction.
Figure 4
Figure 4
Response surface plot illustrates the effect of LR:SC ratio (X1) and incubation time (X3) on % AFM1 reduction.
Figure 5
Figure 5
Response surface plot illustrates the effect of incubation temperature (X2) and incubation time (X3) on % AFM1 reduction.
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