Table 3.

Summary of the main findings of the studies on the effect of microbes and enzymes on aflatoxin (AF) mitigation.

Microbes and Enzymes		In Vitro		In Vivo					Animal Health Status/Zootechnical Parameters	Other Experiments	Remarks
		AF Adsorption/Binding	Antifungal Activity AF Inhibition	AF Degradation/Detoxification in Feed	AF Degradation/Reduction in Animals 1	Extinguishing AF Immunomodulation	Extinguishing AF Genotoxic Effect	Extinguishing AF Cytotoxic Effect
Dogi et al. (2017) [73]	Saccharomyces cerevisiae RC016								+	AFB1 effect on S. cerevisiae cells—significant increase in cell diameter
Gonzales Pereyra et al. (2014) [74]	Saccharomyces cerevisiae RC016						++	0	0/+
Magnoli et al. (2016) [56]	Clavispora lusitaniae, Pichia kudriavzevii, Cyberlindnera fabianii, Candida tropicalis	+								AFB1 desorption study—irreversible binding was shown	All the tested strains were able to bind AFB1; however, the highest AFB1 affinity was observed for Cl. lusitaniae from feedstuff and the lowest value was observed for P. kudriavzevii from feedstuff. Cy. fabianii isolated from faeces and Ca. tropicalis isolated from the gut showed moderate affinity
Poloni et al. (2015) [75]	Saccharomyces cerevisiae strains RC009	0									Potentiation of a feed additive premix by different strains was investigated
Poloni et al. (2015) [75]	Saccharomyces cerevisiae strains RC012	++									Potentiation of a feed additive premix by different strains was investigated
Poloni et al. (2015) [75]	Saccharomyces cerevisiae strains RC016	++									Potentiation of a feed additive premix by different strains was investigated
Gonçalves et al. (2017) [76]	Saccharomyces cerevisiae				++				0		S. cerevisiae types studied were cell wall, dried yeast, autolyzed yeast and brewery yeast. Cell wall and autolyzed yeast showed the best results for aflatoxin reduction
Tayel et al. (2013) [68]	Pichia anomala ATCC 34080		+						+	Hydrolytic enzyme secretion experiment—β-1,3-glucanase and exo-chitinase activity
Dogi et al. (2015) [62]	Lactobacillus rhamnosus RC007 and Lactobacillus plantarum RC009		++								L. rhamnosus RC007 was the most efficient at inhibiting the three fungal species
Zielinska and Fabiszewska (2018) [63]	Lactobacillus buchneri A KKP 2047 p, Lactobacillus reuteri M KKP 2048 p, Lactobacillus plantarum K KKP 593 p, Lactobacillus plantarum S KKP 2021 p, Lactobacillus fermentum N KKP 2020		++							Studies relating to the synergistic activity of bacterial strains were also conducted on a production scale. It resulted in a decrease in mould count and a decrease in AFB1 levels in silages by 74% and 75%, respectively, compared to the negative control	The bacterial strains had a synergistic effect and decreased the AFB1 levels by about 80% compared to the control silage and by about 74% compared to the silage inoculated with only one strain (L. buchneri A KKP 2047 p)
Ying et al. (2017) [77]	Lactobacillus rhamnosus			++						Fermentation characteristics, in vitro digestibility—positive effects	Reduction of aflatoxin production in silage was investigated during exposure to air
Ma et al. (2017) [78]	Ten Lactobacillus species	++								Viability and pH studies on binding: the greatest binding of AFB1 within a bacterium was achieved by dead L. plantarum and L. buchneri and viable Pediococcus acidilactici at pH 2.5. Binding efficacy generally decreased in a quadratic manner as the acidity of the culture media decreased	When applied at 109 CFU/mL, all the 10 bacteria bound AFB1, but L. plantarum R2014 (Lp) and EQ12, L. buchneri R1102 (Lb) and Pediococcus acidilactici R2142 and EQ01 (Pa) had the greatest capacity
Drobná et al. (2017) [57]	Lactobacillus reuteri E and Lactobacillus mucosae D, Lactobacillus murinus C, Lactobacillus reuteri KO5, Lactobacillus reuteri KO4b, Lactobacillus reuteri KO4m, Lactobacillus plantarum KG1, Lactobacillus plantarum KG4	++	++							pH studies—the highest inhibition of fungal growth was observed at pH 4	The highest growth inhibition of A. flavus was shown by L. mucosae D. The best results concerning AFB1 reduction were obtained with the L. reuteri KO4b strain followed by L. plantarum KG4
Rather et al. (2014) [64]	Lactobacillus plantarum YML007		++	++					+
Dogi et al. (2013) [61]	Lactobacillus rhamnosus RC007		++							Antibiotic resistance—no genes for resistance to the tested antibiotics
Dogi et al. (2013) [61]	Lactobacillus plantarum RC009		++		-						Inhibition only at pH 4
Nasrabadi et al. (2013) [79]	Lactobacillus casei Shirota				+			+	++
Jebali et al. (2015) [65]	Lactobacillus plantarum MON03	++				++			++
Zhang et al. (2019) [80]	Lactobacillus rhamnosus GG				++			+			Single dose of AFB1 administration
Ben Salah-Abbés et. al. (2015) [66]	Lactobacillus plantarum MON03	++				++	++	++			Live LP showed better binding percentages than heat-killed LP
Intanoo et al. (2018) [58]	Ruminal fluid isolates—Kluyveromyces marxianus and Pichia kudriavzevii (yeast); Enterococcus faecium, Corynebacterium phoceense and Corynebacterium vitaeruminis (bacteria)			++						Preliminary assessment on biomass production—the isolates could be produced in bulk for their potential use as feed supplements for dairy cattle	The best yeast isolates were identified as K. marxianus and P. kudriavzevii. Generally, yeasts showed better detoxifying performance than bacteria in liquid media and similar but faster detoxification rates in TMR
Wang et al. (2018) [69]	Microbial consortium TMDC (Geobacillus (12.3%), Tepidimicrobium (36.65%), Clostridium III (21.2%), Aeribacillus (8.84%), Cellulosibacter (5.1%), Desulfotomaculum (6.44%) and Tepidanaerobacter (3.14%))			++						Simultaneous degradation of AFB1 and ZEA was studied	Cell-free supernatants, cell pellets and intracellular extracts of TMDC were studied. Supernatants of TMDC played a dominant role in mycotoxin degradation by the microbial consortium. Geobacillus and Tepidimicrobium genera played important roles in mycotoxin degradation
Wang et al. (2019) [59]	Escherichia coli CG1061			++						Temperature studies—the active component might be heat-resistant; pH studies—degradation rates of alkaline conditions were higher than those of acidic conditions; toxicity studies—biotransformed AFB1 was less toxic	The culture supernatant showed a significantly higher degradation rate than that of intracellular extracts
Prettl et al. (2017) [60]	Rhodococcus pyridinivorans K408			++						Biomass growth—changed to a stagnant state after seven days of incubation in harmony with the mycotoxin degradation rate
Xu et al. (2017) [67]	Bacillus shackletonii LMG 18435			++						Enzyme characterization—thermostable enzyme named Bacillus aflatoxin-degrading enzyme (BADE) responsible for AFB1 degradation activity was purified and characterized	The culture supernatant of the tested isolate was more effective than viable cells and cell extracts
Scarpari et al. (2014) [70]	Trametes versicolor TF294, CF294		++	++						AFB1 degradation experiments with the laccase enzyme—significant decrease under in vitro and in vivo conditions (liquid culture and maize). Toxicity study of the AFB1 by-product of the laccase enzyme—no toxic effects were shown
Das et al. (2014) [71]	Pleurotus ostreatus MTCC 142 and Pleurotus ostreatus GHBBF10			++						Effect of metal ions and surfactants on degradation—enhanced degradation was noted for P. ostreatus MTCC 142 in the presence of Cu2+ and Triton X-100 at the toxin concentration of 5 µg/mL. P. ostreatus GHBBF10 showed the highest degradation in the presence of Zn2+ and Tween 80	The highest degradation was recorded for both strains at the 0.5 µg/mL initial concentration of AFB1. With an increase in AFB1 concentration, progressive decrease in degradation was encountered
Branà et al. (2017) [72]	Pleurotus eryngii		++	++						Translocation of AFB1 and aflatoxicol through the thallus to the basidiocarps (fruit bodies)—neither the biomass produced on the mushroom substrate nor the mature basidiocarps contained detectable levels of AFB1 or its metabolite aflatoxicol	The addition of 5% wheat straw to the culture medium increased the tolerance of P. eryngii to AFB1

Legend: ++—significant; +—not significant or not indicated in the study; 0—no change; -—negative effect; empty cell—not examined. Results of the same publication are indicated with a thick frame. ¹ Parameters such as carryover rate, aflatoxin excretion, aflatoxin levels in blood, urine, faeces, milk.