Potential preventive role of lactic acid bacteria against Aflatoxin M₁ immunotoxicity and genotoxicity in mice

Figures & data

Table 1. Effects of simulated gastric juice and bile salt on LAB strains, and Caco-2 cell adherence activity.

	Initial mean	Gastric juice pH		Bile salt level (w/v)				Caco-2 cell
Bacteria	counts^a	pH 2	pH 3	0.3%	0.5%	1%	BSH activity	adhesion^b
L. plantarum MON03	9.46 ± 0.15	9.16 ± 0.55	9.11 ± 0.51	7.14 ± 0.31	7.67 ± 0.32	7.73 ± 0.45	+	457.8 ± 106.2
L. rhamnosus GG	9.61 ± 0.12	9.20 ± 0.42	9.05 ± 0.56	7.14 ± 0.37	7.77 ± 0.41	7.41 ± 0.36	+	489.1 ± 98.5

^aMean number of lactobacilli as log CFU/ml.

^bTotal number of lactobacilli that adhered to Caco-2 cells in 20 random microscopic fields.

Each value represents mean (±SD) from three trials per assay condition.

+, Positive for BSH activity.

Table 2. Percentage AFM₁ removal in PBS by the presence of LP.

	AFM1 (50 µg/ml)
Treatments	0 h	12 h	24 h
PBS only	3.0 ± 0.2	2.4 ± 0.2	2.3 ± 0.2
Live LP	25.9 ± 5.8*	67.1 ± 6.2*	93.1 ± 5.2*
Heat-killed LP	25.1 ± 2.1	40.5 ± 2.6	55.1 ± 4.5

*Value significantly different from control (PBS-only and heated bacteria) at p < 0.05.

Table 3. Absolute body weight, percentage weight gain, and spleen and thymus weights and indices in mice.

Parameters	Control	LP	AFM1	LP + AFM1
Body weight (g)
initial	26.60 ± 1.20	29.80 ± 2.10	23.50 ± 1.90	31.50 ± 2.10
final	33.80 ± 1.70	42.34 ± 3.20*	26.34 ± 2.50	47.28 ± 4.60
Body weight gain (%)	27.10 ± 3.10	42.10 ± 2.20*	12.10 ± 1.00*	50.10 ± 5.10*
Spleen wet weight (g)	0.18 ± 0.20	0.19 ± 0.30	0.20 ± 0.20	0.22 ± 0.30
Spleen Index	0.53 ± 0.03	0.48 ± 0.04	0.75 ± 0.06*	0.46 ± 0.03
Thymus wet weight (g)	0.014 ± 0.003	0.016 ± 0.002*	0.013 ± 0.001*	0.022 ± 0.002
Thymus Index	0.041 ± 0.002	0.038 ± 0.001	0.049 ± 0.003*	0.046 ± 0.002*

All mice were orally exposed daily for 15 days to LP (10⁹CFU/kg), AFM₁ (100 mg/kg), or a co-treatment with LP + AFM₁ (at the respective doses noted here).

Values shown are mean (±SD) from 10 determinations per experimental scenario tested.

*Value significantly different from control at p ≤ 0.05.

Organ Indices calculated as 100 × [organ weight (g)/total body weight (g)].

Table 4. Effect of LP on total white blood cell levels and leukocyte differentials in mice.

Groups	WBC (×10^3/µl)	NE (×10^3/µl)	LY (×10^3/µl)	MO (×10^3/µl)	EO (×10^3/µl)
Control	8.02 ± 0.13	6.15 ± 0.07	1.41 ± 0.06	0.42 ± 0.09	0.14 ± 0.06
LP	8.11 ± 0.22	6.11 ± 0.07	1.43 ± 0.08	0.44 ± 0.06	0.08 ± 0.02
AFM1	12.13 ± 0.40*	9.74 ± 0.25*	0.84 ± 0.11*	1.86 ± 0.10*	0.65 ± 0.02*
LP + AFM1	8.86 ± 0.12	6.12 ± 0.15	1.36 ± 0.05	0.40 ± 0.21	0.15 ± 0.04

All treatment regimens are as outlined in the legend for Table 3.

Values shown are mean (±SD) from 10 determinations per experimental scenario tested.

*Value significantly different from control at p ≤ 0.05.

Figure 1. Changes in spleen and thymus cellularity. Mice were orally exposed daily to AFM₁ (100 mg/kg BW), LP (10⁹CFU/L, ∼1 mg/kg BW), or AFM₁ + LP for 15 days. Controls received vehicle only each day. Data shown are mean (±SD). ^aValue significantly different from vehicle control (p < 0.05).

Table 5. Mean number of micronuclei in polychromatic erythrocytes and chromosomal aberrations in bone marrow cells of mice.

Treatments	Micronucleus	Total aberrations
Control	1.66 ± 0.27^a	3.59 ± 0.32^a
Colchicine/Mitomycin C	67.21 ± 8.10^c	62.11 ± 5.22^c
LP	2.28 ± 0.45^a	3.45 ± 0.62^a
AFM1	51.27 ± 8.14^c	47.22 ± 4.12^c
LP + AFM1	7.11 ± 0.27^b	6.65 ± 0.81^b

All treatment regimens are as outlined in the legend for Table 3.

Values shown are mean (±SD) from 10 determinations per experimental scenario tested.

^a,b,cWithin each column, values with differing superscripts significantly differ at p < 0.05.