In Vitro Detoxification of Aflatoxin B1, Deoxynivalenol, Fumonisins, T-2 Toxin and Zearalenone by Probiotic Bacteria from Genus Lactobacillus and Saccharomyces cerevisiae Yeast

In Vitro Detoxification of Aflatoxin B1, Deoxynivalenol, Fumonisins, T-2 Toxin and Zearalenone by... The aim of the following research was to determine the detoxification properties of probiotic Lactobacillus sp. bacteria (12 strains) and S. cerevisiae yeast (6 strains) towards mycotoxins, such as aflatoxin B , deoxynivalenol, fumonisins, T-2 toxin and zearalenone, which pose as frequent feed contamination. The experiment involved analysing changes in concentration of mycotoxins in PBS solutions, after 6, 12 and 24 h of incubation with monocultures of tested microorganisms, measured by high-performance liquid chromatography (HPLC). We found that all strains detoxified the mycotoxins, with the highest reduction in concentration observed for the fumonisin B and B mixture, ranging between 62 and 77% for bacterial strains and 67–74% for 1 2 yeast. By contrast, deoxynivalenol was the most resistant mycotoxin: its concentration was reduced by 19–39% by Lactobacillus sp. strains and 22–43% by yeast after 24 h of incubation. High detoxification rates for aflatoxin B , T-2 toxin and zearalenone were also observed, with concentration reduced on average by 60%, 61% and 57% by Lactobacillus,respectively, and65%, 69% and 52% by yeast, respectively. The greatest extent of reduction in the concentration for all mycotoxins was observed after 6 h of incubation; however, a decrease in concentration was noted even after 24 h of incubation. Thus, the tested microorganisms can potentially be used as additives to decrease the concentrations of toxins in animal feed. . . . . Keywords Mycotoxins Detoxification Probiotics Lactobacillus Saccharomyces cerevisiae Introduction fumonisins (FUM), along with zearalenone (ZEN) are the most prevalent mycotoxin-related contamination found in Mycotoxins are secondary metabolites with low molecular fodder [5, 6]. AFB is most frequently found in feed in the mass (~ 700 Da), which are synthesised by filamentous fungi, European Union (> 98% of tested samples); however, DON belongingmostlytothe Ascomycota phylum. The most com- (~ 90% of tested samples) and ZEN (~ 70% of tested samples) mon source of food and feed contamination are mycotoxins are often detected as well. The presence of FUM and OTA, on produced by the fungi Aspergillus, Penicillium and Fusarium the other hand, are more sparsely observed [7]. genera [1–3]. Other mycotoxin-producing fungi include Plants are contaminated with mycotoxins, synthesised by Alternaria, Chaetomium, Cladosporium, Claviceps, filamentous fungi, most frequently at the time of cultivation in Diplodia, Myrothecium, Monascus, Phoma, Phomopsis, the fields (e.g. mycotoxin produced mostly by Fusarium sp.). Pithomyces, Trichoderma and Stachybotrys [4]. Aflatoxins Likewise, under favourable growth conditions of temperature (AF), ochratoxin (OT), trichotecens, including and humidity, mycotoxin-producing fungi, such as deoxynivalenol (DON) and T-2 toxin (T-2), as well as Aspergillus sp. and Penicillium sp., are also found in food and feed that are stored [2, 8, 9]. In stored grains, moisture content within 16–30%, high temperature reaching 25–30 °C, * Agnieszka Chlebicz and high relative air humidity (80–100%) are conditions that agnieszka.chlebicz@edu.p.lodz.pl stimulate growth of filamentous fungi and mycotoxin produc- * Katarzyna Śliżewska tion [10]. The concentration of toxins in input materials (e.g. katarzyna.slizewska@p.lodz.pl corn, grass, clover) is not reduced to a sufficient degree while they are being processed into feed, as these metabolites are Institute of Fermentation Technology and Microbiology, Department resistant to high and low temperatures, even after long storage of Biotechnology and Food Sciences, Lodz University of Technology, Wólczańska 171/173, 90-924 Łódź, Poland period [9, 11]. Therefore, these toxins constitute a threat, as Probiotics & Antimicro. Prot. they can enter the human food chain through products such as mycotoxins to less toxic metabolites, and the proper treatment milk, meat or eggs [2]. Furthermore, humans are exposed to of food or feed by fermentation are also used for biological mycotoxin-related intoxications while consuming foods of detoxification [9, 20–23]. In comparison to physical and plant origin, for instance hazelnuts, almonds, grains and fruits chemical methods, biological detoxification is more efficient, [8]. Therefore, European Union legislation specifies tolerable specific and safer for the environment [22]. daily intake (TDI) for a variety of mycotoxins, in addition to In 1998, El-Nazami performed pioneering in vitro studies providing guidance values for their concentrations in animal on the binding properties of mycotoxin by lactic acid bacteria, feedstuffs (Table 1)[18]. which has initiated a systematic search for microorganisms The regulation (EU) 2017/625 of the European Parliament having specific abilities to adsorb mycotoxins [24]. Among and of the Council of 15 March 2017, regarding official con- other microbes identified for biological detoxification pur- trols and other official activities performed to ensure the ap- poses, probiotic microorganisms, defined by FAO/WHO plication of food and feed law, rules on animal health and (2002) as ‘live microorganisms, which when administered in welfare, plant health and plant protection products will come adequate amounts confer a health benefit on the host’,were into effect on 14th December 2019. According to this regula- identified as microbes that bind and adsorb mycotoxins [25]. tion, Member States are obliged to establish multiannual plans Probiotic microorganisms, such as bacteria belonging to gen- and carry out food and feed controls, to ensure safety in the era Lactobacillus, Bifidobacterium,as well as Enterococcus agri-food chain, as well as animal welfare and health, thereby faecium,andtheyeasts Saccharomyces cerevisiae and providing safe food [19]. Saccharomyces boulardii have been shown to have mycotox- The process of detoxification or mycotoxin removal is in detoxification properties [26]. complicated, especially because of heat stability of these com- The aim of our study was to determine detoxification prop- pounds, and their breakdown into toxic products. erties of probiotic strains of Lactobacillus sp. and S. cerevisiae Nevertheless, detoxification may be accomplished by applica- towards mycotoxins, which often contaminate feed for live- tion of the following methods: physical (cooking, baking, mi- stock animals. This study was part of one of the stages of crowave heating, radiation, etc.), chemical (use of ammonia, strain selection for designing synbiotic preparations for poul- hydrochloric acid, salicylic, sulfamide, sulfosalicylic, try and swine. anthranilic, benzoic, boric, oxalic or propionic acid), and bio- logical [20]. For biological detoxification, plant extracts, such as piperine, lutein, carotenoids or essential oils, as well as Materials and Methods enzymes, such as AF decomposing peroxidase or laccase, and FUM degradative carboxylesterase or aminotransferase Biological Materials are used. Microorganisms that are capable of degrading The biological material included potential probiotic bacteria of Lactobacillus genus and strains of the yeast S. cerevisiae, Table 1 Optimal conditions for mycotoxins production, TDI in food which are deposited in the Łódź Collection of Pure Cultures products and guidance value in feedstuff in European Union [12–17] 105 of Institute of Fermentation Technology and Mycotoxin TDI in Guidance Optimum Optimum Microbiology at Technical University of Łódź (Table 2). food value in temperature water activity Five Lactobacillus (rhamnosus ŁOCK 1087, paracasei (μg/kg)* feedstuff (12% for mycotoxin for ŁOCK 1091, reuteri ŁOCK 1092, plantarum ŁOCK 0860, moisture) production mycotoxin pentosus ŁOCK 1094) and one S. cerevisiae (ŁOCK 0119) (mg/kg)** (°C) production strain have been documented in patent application no. 422603 AF*** 0.025–15.0 0.005–0.05 33 0.99 [27]. DON 200–1750 0.9–12 26–30 0.995 The detoxification activity of bacteria and yeast was deter- FUM**** 200–2000 5–60 15–30 0.9–0.995 mined against five mycotoxins, namely aflatoxin B1, OTA 0.50–10.0 0.05–0.25 25–30 0.98 fumonisin mixture of fumonisin B1 and B2 (FUM), T-2 toxin, T-2/HT-2 15–1000 0.25–220–30 0.98–0.995 zearalenone and deoxynivalenol (Table 3). Mycotoxins were ZEA 20–200 0.1–325 0.96 suspended in PBS buffer (Calbiochem®, Germany), and so- lutions of 100 μg/ml were prepared. *Depending on a product (e.g. nuts intended for processing or direct consumption, raw grains, milk, dried fruits, spices, infants’ formulas, Bacterial and Yeast Strain Cultivation and Sample processed foods based on cereals, wine, coffee, etc.) Preparation **With a distinction between feed materials and complementary and complete feed mixtures ***Depending on form Lactobacillus sp. were cultivated in de Man, Rogosa, and ****Sum of FB and FB Sharpe (MRS) broth (Merck, Germany) at 37 °C, while 1 2 Probiotics & Antimicro. Prot. Table 2 Strains whose Microorganism Collection number Source of isolation mycotoxin detoxification properties has been studied Bacteria Lact. brevis ŁOCK 1093 Plant silages Lact. casei ŁOCK 0911 Milk fermented beverages Lact. casei ŁOCK 0915 Milk fermented beverages Lact. paracasei ŁOCK 1091 Caecal content of sow Lact. pentosus ŁOCK 1094 Broiler chicken dung Lact. plantarum ŁOCK 0860 Plant silages Lact. plantarum ŁOCK 0862 Plant silages Lact. reuteri ŁOCK 1092 Piglet caecal content Lact. reuteri ŁOCK 1096 Winear pig’s intestinal content Lact. rhamnosus ŁOCK 1087 Turkey dung Lact. rhamnosus ŁOCK 1088 Broiler chicken’s intestinal content Lact. rhamnosus ŁOCK 1089 Broiler chicken’s intestinal content Yeast S. cerevisiae ŁOCK 0068 Forage S. cerevisiae ŁOCK 0113 Distillers’ yeast, potato with grain S. cerevisiae ŁOCK 0119 Distillers’ yeast, grain S. cerevisiae ŁOCK 0137 Baker’syeast S. cerevisiae ŁOCK 0140 Baker’syeast S. cerevisiae ŁOCK 0142 Baker’syeast S. cerevisiae was grown at 30 °C, in yeast extract–peptone– was used with a 250 × 4.6 mm size ACE 5 C18 column (Advanced glucose broth (YPG Broth, Merck, Germany). Both bacteria Chromatography Technologies (ACT), Scotland). Mycotoxins and yeast cultures were grown in normal oxygen conditions were identified by comparing the retention times of the peak with for 24 h. After 24 h of incubation, monocultures of the standard solutions. The mycotoxin concentrations were deter- analysed strains, in three repetitions, were centrifuged at rela- mined by correlation of peak area of the samples with the stan- tive centrifugal force (RCF) 3468×g for 10 min (Centrifuge dard curves, obtained by HPLC analysis of standard solutions. MPW-251; MPW, Poland). Subsequently, the supernatants were removed and the bacteria and yeast biomass were Statistical Analysis washed three times with PBS buffer to remove any residual culture medium. The cell pellets were again centrifuged under The results presented here constitute the arithmetic mean of the same conditions. Ten millilitres of mycotoxin solutions values from three repetitions, with standard deviation. All sta- were added to the prepared samples, with a defined concen- tistical analyses were carried out using the one-way ANOVA tration of 100 μg/ml for each mycotoxin. These samples were test, with a significance level of p < 0.05 (Origin 6.1 program, further incubated in normal oxygen conditions for 24 h at OriginLab). A comparative Duncan test was carried out at a 37 °C or 30 °C for lactic acid bacteria and yeast, respectively. significance level of p > 0.05 (STATISTICA 10, StatSoft). After 6, 12 and 24 h of incubation, 2 ml of each sample was collected, centrifuged at RCF of 3468×g for 10 min, and the supernatants were filtered with PTFE syringe filters with Results 0.22-μm-diameter pores (Millex-GS, Millipore, USA). As a positive control sample, a solution of analysed mycotoxin in AFB Microbial Detoxification PBS was used, and bacterial or yeast suspension served as negative control sample. Bacteria belonging to the Lactobacillus genus were characterised by their diverse ability to detoxify aflatoxin HPLC Analysis B . In merely after 6 h of incubation, a statistically significant reduction of AFB concentration was noticed, ranging from The prepared samples were subjected to high-performance liquid 35.33 to 79.65 μg/ml (20–65% reduction, 49% on average) chromatography (HPLC) analysis, the parameters of which are compared to the initial mycotoxin concentration of 100 μg/ml. presented in Table 4. Analysis was performed as previously de- In subsequent hours, further reduction in AFB concentration scribed by El-Nazami et al. [24], with modifications. For this pur- was observed. After 24 h of incubation, the concentration of pose, Surveyor liquid chromatography (Thermo Scientific, USA) mycotoxin in the samples was 28.96–55.80 μg/ml (mean, Probiotics & Antimicro. Prot. Table 3 Mycotoxins, that were detoxified by selected strains of potentially probiotic microorganisms (Sigma-Aldrich, available online on https://www. sigmaaldrich.com/, accessed on 21 March 2018 [28–33]) Mycotoxin Chemical structure Producer, catalogue number Producer, catalogue number of HPLC sample AFB Sigma, A6636 Supelco, 46323-u DON Sigma, 32943 Supelco, CRM46911 FUM mixture Sigma, 34143 Fumonisin B – Sigma, 34139 Fumonisin B Fumonisin B – Sigma, 34142 Fumonisin B T-2 Sigma, T4887 Sigma, 34071 ZEA Sigma, Z2125 Supelco, CRM46916 Probiotics & Antimicro. Prot. Table 4 HPLC analysis parameters Parameter Mycotoxins AFB DON FUM T-2 ZEN Column heating – 30 –– – Mobile phase Water/acetonitrile/methanol Water/acetonitrile Gradient methanol/water Methanol/water Methanol/water (60:30:10) (90:10) (70:30 and 80:20) (60:40) (70:30) Fluorescent detector λ (nm) (ex- 360 and 420 – 490 and 450 381 and 470 280 and 460 citation and emission) UV detector λ (nm) – 218 –– – Flow (ml/min) 1 1 1 1 1 40.43 μg/ml). Therefore, there was a reduction of between 44 DON Microbial Detoxification and 71% (mean, 60%) compared to the initial concentration of the mycotoxin. Deoxynivalenol (DON) concentrations were significantly re- S. cerevisiae showed detoxification activity similar to that duced after 6 h of incubation in the presence of the bacteria of analysed strains of Lactobacillus sp. After 6hof incuba- monoculture, varying between 78.39 and 94.24 μg/ml (mean, tion, AFB concentrations were statistically significantly re- 84.30 μg/ml), indicating that these bacteria have the ability to duced by 47–66% (average 58%) and ranged from 33.64 to decrease DON concentration by an average of 16%. Further 53.27 μg/ml. In subsequent hours of incubation, the concen- decrease in DON concentration in the subsequent incubation tration of the mycotoxin further decreased, and after 24 h, the hours was also observed. After 24 h, DON concentrations mycotoxin concentrations were 32.48–4.45 μg/ml (average ranged between 60.63 and 80.72 μg/ml (mean, 70.45 μg/ml), reduction of AFB1 by 65%) (Table 5). thereby showing a reduction of 19–39% (mean, 30%). Table 5 Reduction of AFB concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B B C Lactobacillus brevis ŁOCK 1093 100 39.56 ± 0.99 (60) 37.13 ± 2.04 (63) 32.70 ± 0.59 (67) B B C casei ŁOCK 0911 51.30 ± 2.04 (49) 49.92 ± 1.36 (50) 45.13 ± 0.38 (55) B C D casei ŁOCK 0915 67.18 ± 0.63 (33) 58.98 ± 1.27 (41) 55.80 ± 1.51 (44) B C D paracasei ŁOCK 1091 57.44 ± 1.61 (43) 48.23 ± 1.27 (52) 42.21 ± 1.52 (58) B B B pentosus ŁOCK 1094 44.70 ± 0.41 (55) 38.73 ± 1.03 (61) 28.96 ± 0.58 (71) B B C plantarum ŁOCK 0860 47.93 ± 1.01 (52) 45.88 ± 0.88 (54) 40.62 ± 0.50 (59) B B B plantarum ŁOCK 0862 35.33 ± 0.95 (65) 34.82 ± 0.47 (65) 34.30 ± 1.06 (66) B B B reuteri ŁOCK 1092 45.19 ± 0.97 (55) 44.98 ± 0.60 (55) 43.79 ± 1.81 (56) B C D reuteri ŁOCK 1096 40.79 ± 0.74 (59) 38.41 ± 1.08 (62) 36.02 ± 0.57 (64) B B B rhamnosus ŁOCK 1087 41.42 ± 2.34 (59) 40.32 ± 0.34 (60) 40.19 ± 1.48 (60) B C D rhamnosus ŁOCK 1088 79.65 ± 0.96 (20) 61.04 ± 0.88 (39) 44.83 ± 1.49 (55) B C D rhamnosus ŁOCK 1089 56.24 ± 0.98 (44) 50.43 ± 0.46 (50) 40.61 ± 1.58 (59) Average concentration (μg/ml) (decrease (%)) 50.56 (49) 45.74 (54) 40.43 (60) B B C S. cerevisiae ŁOCK 0068 100 A 46.79 ± 0.62 (53) 44.43 ± 1.35 (56) 38.07 ± 1.35 (62) B C D ŁOCK 0113 53.27 ± 0.69 (47) 48.93 ± 1.75 (51) 41.45 ± 0.63 (59) B C D ŁOCK 0119 42.25 ± 0.73 (58) 34.12 ± 1.05 (66) 31.48 ± 1.05 (69) B B B ŁOCK 0137 34.99 ± 1.85 (65) 34.02 ± 2.30 (66) 32.70 ± 0.79 (67) B B C ŁOCK 0140 41.55 ± 1.62 (58) 39.58 ± 1.42 (60) 35.78 ± 1.39 (64) B B,C C ŁOCK 0142 33.64 ± 0.48 (66) 33.00 ± 1.41 (67) 32.48 ± 0.38 (68) Average concentration (μg/ml) (decrease (%)) 42.08 (58) 39.01 (61) 35.33 (65) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) Probiotics & Antimicro. Prot. S. cerevisiae strains used in the analysis also demonstrated 25.53 and 32.57 μg/ml, demonstrating an average reduction the ability to reduce the concentration of DON in suspension. of 72% of the initial concentration (Table 7). After 6 h, significant reductions of DON concentrations were observed, ranging between 12 and 22% (mean, 18%) of the initial concentration of DON. After a further 6 h, the concen- T-2 Microbial Detoxification trations were 65.57–80.81 μg/ml (mean, 73.20 μg/ml). After 24 h of incubation, the DON concentrations were reduced by After 6 h of incubation, significant reduction of T-2 toxin 22–40% (mean, 33%) relative to the initial concentration of concentrations by monocultures of analysed Lactobacillus mycotoxin (Table 6). sp. strains were observed ranging between 48.25 and 73.32 μg/ml (mean, 57.36 μg/ml). Continued incubation caused a further decrease in T-2 concentration, as a result of FUM Microbial Detoxification which after 24 h of incubation the concentrations of mycotox- in were 31.09–50.10 μg/ml (mean, 39.01 μg/ml). This shift in Bacteria belonging to Lactobacillus genus detoxified the mix- concentration values indicated a reduction of 50–69% (mean, ture of fumonisin B (FB )and B (FB )mycotoxins (FUMs). 1 1 2 2 61%) in relation to the initial quantity of T-2. After 6 h of incubation, the FUM concentration was reduced S. cerevisiae strains subjected to the analysis were by 36–64% (mean, 51%) compared to the initial concentration characterised by diverse T-2 detoxification activity. After 6-h of the mycotoxin mixture. Subsequent incubation resulted in a incubation, a statistically significant decrease in the concen- further significant decrease in FUM concentration, which after tration of T-2 to level of 46.92–54.98 μg/ml (average reduc- 24 h of incubation was reduced by an average of 70% and tion of 49% of initial concentration) was observed. In subse- ranged from 23.08 to 38.42 μg/ml (mean, 29.53 μg/ml). quent hours of incubation, T-2 concentration further de- S. cerevisiae also reduced the concentrations of FUM, sig- creased. The concentration of the mycotoxin after 24 h of nificantly reducing them by 29–60% (mean, 53%) to 40.15– incubation reduced by 60–63% (mean, 61%) of the initial 70.57 μg/ml (mean, 47.19 μg/ml). In subsequent hours of concentration and ranged between 37.36 and 40.40 μg/ml incubation, the concentrations of mycotoxin further declined. (mean, 38.68 μg/ml) (Table 8). Finally, after 24 h, the FUM concentration ranged between Table 6 Reduction of DON concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B C D Lactobacillus brevis ŁOCK 1093 100 94.24 ± 0.77 (6) 87.52 ± 0.95 (12) 80.72 ± 0.33 (19) B C D casei ŁOCK 0911 92.06 ± 1.10 (8) 84.06 ± 1.01 (16) 72.49 ± 0.67 (28) B C D casei ŁOCK 0915 88.73 ± 1.85 (11) 81.93 ± 0.75 (18) 78.06 ± 1.20 (22) B C D paracasei ŁOCK 1091 84.44 ± 1.75 (16) 73.75 ± 3.09 (26) 67.30 ± 1.46 (33) B C D pentosus ŁOCK 1094 78.53 ± 1.19 (21) 72.49 ± 1.88 (27) 66.82 ± 0.65 (33) B C D plantarum ŁOCK 0860 80.34 ± 0.49 (20) 74.12 ± 0.83 (26) 70.25 ± 1.01 (30) B C D plantarum ŁOCK 0862 83.78 ± 0.20 (16) 78.34 ± 1.27 (22) 74.53 ± 1.04 (25) B C D reuteri ŁOCK 1092 79.92 ± 1.02 (20) 69.05 ± 0.44 (31) 60.63 ± 0.59 (39) B C D reuteri ŁOCK 1096 78.36 ± 0.30 (22) 70.58 ± 1.38 (29) 61.31 ± 1.45 (39) B C D rhamnosus ŁOCK 1087 87.85 ± 2.94 (12) 79.04 ± 1.13 (21) 75.05 ± 1.43 (25) B C D rhamnosus ŁOCK 1088 83.04 ± 2.78 (17) 78.14 ± 0.86 (22) 73.74 ± 0.30 (26) B C D rhamnosus ŁOCK 1089 80.35 ± 0.49 (20) 74.01 ± 2.61 (26) 64.55 ± 0.70 (35) Average concentration (μg/ml) (decrease (%)) 84.30 (16) 76.92 (23) 70.45 (30) B C D S. cerevisiae ŁOCK 0068 100 A 77.68 ± 1.53 (22) 70.62 ± 1.17 (29) 64.57 ± 0.57 (35) B C D ŁOCK 0113 88.32 ± 0.88 (12) 80.81 ± 1.45 (19) 78.03 ± 0.82 (22) B C D ŁOCK 0119 80.87 ± 0.85 (19) 65.57 ± 1.52 (34) 57.50 ± 0.83 (43) B B C ŁOCK 0137 84.16 ± 1.40 (16) 80.41 ± 2.27 (20) 76.10 ± 1.32 (24) B C D ŁOCK 0140 80.64 ± 1.71 (19) 71.97 ± 2.04 (20) 63.07 ± 1.49 (37) B C D ŁOCK 0142 80.28 ± 2.07 (20) 69.80 ± 1.61 (30) 60.91 ± 1.24 (39) Average concentration (μg/ml) (decrease (%)) 81.99 (18) 73.20 (27) 66.70 (33) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) Probiotics & Antimicro. Prot. Table 7 Reduction of FUM concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B C D Lactobacillus brevis ŁOCK 1093 100 50.14 ± 1.26 (50) 43.87 ± 1.01 (56) 34.27 ± 1.26 (66) B C C casei ŁOCK 0911 47.80 ± 3.20 (52) 40.63 ± 0.46 (59) 36.32 ± 3.27 (64) B C D casei ŁOCK 0915 48.52 ± 3.21 (51) 41.98 ± 1.81 (58) 33.50 ± 1.11 (67) B C D paracasei ŁOCK 1091 55.98 ± 3.55 (44) 45.56 ± 1.57 (54) 38.42 ± 1.39 (62) B C D pentosus ŁOCK 1094 45.24 ± 0.41 (55) 38.61 ± 1.02 (61) 29.61 ± 1.04 (70) B C D plantarum ŁOCK 0860 35.52 ± 1.92 (64) 30.88 ± 0.69 (70) 25.11 ± 1.67 (75) B C D plantarum ŁOCK 0862 38.72 ± 1.04 (61) 30.54 ± 0.08 (69) 23.19 ± 1.60 (77) B C D reuteri ŁOCK 1092 42.64 ± 1.18 (57) 37.38 ± 1.34 (63) 27.23 ± 1.17 (73) B C D reuteri ŁOCK 1096 41.83 ± 1.20 (58) 34.62 ± 2.32 (65) 28.22 ± 1.05 (72) B C D rhamnosus ŁOCK 1087 56.53 ± 2.13 (43) 45.39 ± 0.36 (55) 24.00 ± 0.69 (76) B C D rhamnosus ŁOCK 1088 63.64 ± 1.98 (36) 53.87 ± 1.27 (46) 31.43 ± 0.95 (69) B C D rhamnosus ŁOCK 1089 58.75 ± 1.35 (41) 38.67 ± 1.08 (61) 23.08 ± 0.08 (77) Average concentration (μg/ml) (decrease (%)) 48.77 (51) 40.17 (60) 29.53 (70) A B C D S. cerevisiae ŁOCK 0068 100 70.57 ± 0.64 (29) 56.03 ± 1.13 (44) 32.57 ± 2.21 (67) B C D ŁOCK 0113 48.73 ± 0.32 (51) 40.12 ± 1.08 (60) 27.12 ± 1.06 (73) B C D ŁOCK 0119 54.98 ± 1.73 (45) 43.91 ± 1.15 (56) 26.96 ± 1.00 (73) B C D ŁOCK 0137 40.15 ± 1.07 (50) 34.38 ± 2.11 (66) 26.88 ± 1.54 (73) B C D ŁOCK 0140 58.13 ± 0.86 (42) 47.09 ± 1.84 (53) 29.29 ± 1.50 (71) B C D ŁOCK 0142 44.29 ± 0.97 (56) 37.41 ± 0.76 (63) 25.53 ± 0.65 (74) Average concentration (μg/ml) (decrease (%)) 47.19 (53) 43.16 (57) 28.06 (72) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) ZEN Detoxification the highest detoxification activity towards FUM. The most resistant mycotoxin was DON. Lactobacillus sp. showed varied ability for zearalenone (ZEN) We found that the detoxification of AFB , DON, FUM and detoxification. After only 6 h of incubation, a statistically sig- ZEN mycotoxins by the tested Lactobacillus sp. and nificant reduction of ZEN concentration was observed, which S. cerevisiae strains were similar and did not show significant ranged between 28 and 59% (mean, 43%) relative to the initial differences. In contrast, the T-2 compound was more suscep- mycotoxin concentration of 100 μg/ml. In subsequent hours tible to removal from the mixture by yeast monocultures at a of incubation, a further reduction in the mycotoxin concentra- significance level of p < 0.05comparedtobacterial tion was noted. After 24 h of incubation, the concentration of monocultures. ZEN was 27.39–60.05 μg/ml (mean, 40.43 μg/ml). Therefore, The analysis allowed selection of four bacterial strains and a reduction of 40–73% (mean, 57%) in relation to the initial two yeast strains characterised by the best detoxification ca- concentration of the ZEN was observed in monocultures in- pabilities of all analysed mycotoxins. These include Lact. cubated with Lactobacillus sp. rhamnosus ŁOCK 1087, Lact. reuteri ŁOCK 1092, Lact. S. cerevisiae also demonstrated the ability to reduce the plantarum ŁOCK 0860, Lact. pentosus ŁOCK 1094, concentration of ZEN, and after 6 h of incubation, a significant S. cerevisiae ŁOCK 0119 and S. cerevisiae ŁOCK 1042. decrease of 24–42% (average 34%) to level of 57.64– 75.60 μg/ml (mean, 65.55 μg/ml) was observed. In subse- quent hours of incubation, the concentration of the mycotoxin further reduced, and after 24 h, ZEN concentrations ranged Discussion between 41.88 and 55.84 μg/ml (average reduction of 52% of the initial concentration) (Table 9). Mycotoxin detoxification methods can be classified by modes Based on these results, we concluded that mycotoxin of action applied before or after harvesting raw plant mate- (AFB ,DON, FUM,T-2,ZEN) detoxification properties of rials, which are used for human and animal nutrition. During Lactobacillus sp. and S. cerevisiae were strain-dependent. tillage process of plants, good agricultural practice (GAP) Both bacteria and yeast strains, subjected to analysis, showed should be maintained, including crop rotation, cultivation of Probiotics & Antimicro. Prot. Table 8 Reduction of T-2 concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B C D Lactobacillus brevis ŁOCK 1093 100 73.32 ± 3.18 (27) 60.74 ± 0.41 (39) 50.10 ± 1.32 (50) B C D casei ŁOCK 0911 54.11 ± 1.21 (46) 45.04 ± 1.59 (55) 36.83 ± 0.46 (63) B C D casei ŁOCK 0915 51.86 ± 3.06 (48) 43.24 ± 0.96 (57) 35.90 ± 0.48 (64) B C D paracasei ŁOCK 1091 60.17 ± 1.62 (40) 53.57 ± 1.83 (46) 48.24 ± 0.83 (52) B C D pentosus ŁOCK 1094 59.06 ± 2.45 (41) 43.47 ± 1.43 (57) 39.24 ± 1.41 (61) B C D plantarum ŁOCK 0860 48.62 ± 0.03 (52) 39.08 ± 0.17 (61) 31.46 ± 0.71 (69) B C D plantarum ŁOCK 0862 61.76 ± 3.04 (38) 48.28 ± 1.48 (52) 42.71 ± 1.07 (57) B C D reuteri ŁOCK 1092 53.86 ± 3.18 (46) 39.99 ± 1.20 (60) 31.09 ± 0.69 (69) B C D reuteri ŁOCK 1096 55.93 ± 3.57 (44) 42.39 ± 1.11 (58) 36.41 ± 0.72 (64) B C D rhamnosus ŁOCK 1087 48.25 ± 1.24 (52) 38.18 ± 1.37 (62) 26.76 ± 1.30 (73) B C D rhamnosus ŁOCK 1088 58.30 ± 4.13 (52) 49.88 ± 1.20 (51) 38.10 ± 1.42 (62) B C D rhamnosus ŁOCK 1089 63.08 ± 1.45 (37) 51.25 ± 1.99 (49) 45.78 ± 0.56 (54) Average concentration (μg/ml) (decrease (%)) 57.36 (43) 46.66 (54) 39.01 (61) A B C D S. cerevisiae ŁOCK 0068 100 53.66 ± 1.37 (46) 38.96 ± 2.43 (61) 31.83 ± 1.64 (68) B C D ŁOCK 0113 47.78 ± 1.34 (52) 40.04 ± 1.44 (60) 32.25 ± 0.77 (68) B C D ŁOCK 0119 46.92 ± 2.32 (53) 37.57 ± 0.53 (62) 31.06 ± 1.23 (69) B C D ŁOCK 0137 51.51 ± 1.34 (49) 37.73 ± 1.30 (62) 27.71 ± 1.40 (72) B C D ŁOCK 0140 54.98 ± 1.56 (45) 40.44 ± 1.75 (60) 32.62 ± 0.63 (67) B C D ŁOCK 0142 52.23 ± 0.92 (48) 37.36 ± 0.67 (62) 29.48 ± 0.86 (71) Average concentration (μg/ml) (decrease (%)) 51.18 (49) 38.68 (61) 30.54 (69) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) resistant plants, ploughing, irrigation, chemical and biological The AFB concentration in PBS solution was reduced by control of plant diseases and proper use of chemicals (e.g. the tested strains of bacteria to varying degrees, on average fungicides) [22, 34, 35]. When crops have already been har- 49.44% after the first 6 h of incubation, by another 4.82% vested, mycotoxin concentration can be reduced by adjust- during the next 6 h of incubation, and maintained within a ment of appropriate storage conditions (i.e. humidity and tem- range of 44.20–71.04 μg/ml (average reduction of 59.97%) perature), or by using detoxification treatments (physical, after 24 h of incubation. The greatest amount of AFB was chemical, biological) that can degrade, inactivate or decrease bound by bacteria strains after 6 h of incubation, suggesting the toxicity level of mycotoxins and ensure the nutritional that the adsorption of the mycotoxin by Lactobacillus is an value of food. Simultaneously, these detoxification methods immediate process. Haskard et al. (2001) tested the ability of should not introduce any major changes in production process eight Lactobacillus strains to bind AFB to bacterial surfaces technology [9, 34, 35]. using ELISA; their results were similar to ours [39]. Among other mycotoxin detoxification methods, microor- Liew et al. (2018), who also performed AFB binding as- ganisms, inter alia probiotic strains of Lactobacillus sp. and says with ELISA, confirmed results obtained by Haskard et al. S. cerevisiae, are used in case of mycotoxin contamination of (2001), and observed that live cells of Lactobacillus casei food and fodder [36]. Lactobacillus sp. are able to bind myco- Shirota were more efficient in binding mycotoxin, than heat- toxins mostly to cell wall peptidoglycans, polysaccharides and treated organisms [40]. Hernandez-Mendoza et al. (2009), teichoic acid, primarily through hydrophobic interactions, Huang et al. (2017) and Kumar et al. (2018) in their in vitro whereas S. cerevisiae are bind toxic metabolites of filamentous analysis using HPLC also showed the variable range of detox- fungi to the cell wall. In addition, microorganisms biodegrade ification level of AFB by Lactobacillus sp. In these studies, mycotoxins that prevent adsorption of these components inside AFB was bound by the tested strains of bacteria in the range the intestines on animals that feed on the food [24, 37, 38]. of 14–49%, 20.88–59.44% and 0–85%, respectively [41–43]. In this article, in vitro results demonstrated the ability to The variable binding ability of the Lactobacillus strains to reduce the concentration of mycotoxins: AFB , DON, FUM, AFB could be the result of differences in cell wall structure, 1 1 T-2, ZEN in PBS solution by 12 strains of bacteria from especially in terms of teichoic acid and peptidoglycan content Lactobacillus genus, and 6 S. cerevisiae strains. [44]. On the basis of the studies conducted, Gratz et al. (2005) Probiotics & Antimicro. Prot. Table 9 Reduction of ZEN concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B C D Lactobacillus brevis ŁOCK 1093 100 63.61 ± 3.73 (36) 51.71 ± 2.57 (48) 43.17 ± 1.36 (57) B C D casei ŁOCK 0911 64.77 ± 3.78 (35) 57.41 ± 1.95 (43) 49.91 ± 2.33 (50) B C C casei ŁOCK 0915 63.69 ± 2.31 (36) 55.07 ± 1.82 (45) 51.22 ± 2.54 (49) B B C paracasei ŁOCK 1091 55.86 ± 2.35 (44) 50.67 ± 2.33 (49) 45.97 ± 1.38 (54) B B B pentosus ŁOCK 1094 41.21 ± 1.90 (59) 36.26 ± 3.88 (63) 32.22 ± 1.47 (67) B C D plantarum ŁOCK 0860 44.36 ± 1.29 (56) 35.42 ± 0.93 (65) 27.39 ± 1.46 (73) B C D plantarum ŁOCK 0862 69.93 ± 1.29 (30) 62.47 ± 1.73 (38) 56.63 ± 1.21 (43) B C D reuteri ŁOCK 1092 47.48 ± 1.04 (53) 38.26 ± 0.81 (62) 32.84 ± 1.25 (67) B B C reuteri ŁOCK 1096 72.45 ± 3.98 (28) 65.37 ± 2.31 (35) 60.06 ± 0.64 (40) B C D rhamnosus ŁOCK 1087 60.66 ± 1.78 (39) 52.93 ± 2.36 (47) 45.64 ± 1.54 (54) B C D rhamnosus ŁOCK 1088 58.00 ± 3.42 (42) 48.56 ± 0.65 (51) 41.25 ± 1.57 (58) B C D rhamnosus ŁOCK 1089 45.17 ± 1.70 (55) 35.05 ± 1.38 (65) 30.84 ± 0.82 (69) Average concentration (μg/ml) (decrease (%)) 57.27 (43) 49.10 (51) 43.10 (57) A B C D S. cerevisiae ŁOCK 0068 100 75.60 ± 2.88 (24) 55.78 ± 2.10 (44) 49.99 ± 1.25 (50) B C D ŁOCK 0113 68.47 ± 1.52 (32) 62.14 ± 1.80 (38) 55.84 ± 2.72 (44) B C D ŁOCK 0119 57.64 ± 1.67 (42) 47.16 ± 1.61 (53) 41.88 ± 1.21 (58) B C C ŁOCK 0137 58.95 ± 1.68 (41) 50.70 ± 2.14 (49) 47.47 ± 1.18 (53) B C D ŁOCK 0140 68.24 ± 2.23 (32) 55.59 ± 3.32 (44) 47.59 ± 1.31 (52) B C D ŁOCK 0142 64.38 ± 0.80 (36) 52.35 ± 2.83 (48) 46.03 ± 0.26 (54) Average concentration (μg/ml) (decrease (%)) 65.55 (34) 53.95 (46) 48.13 (52) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) showed that the binding of AFB by Lactobacillus sp. is a detoxification activity of 137 Lactobacillus strains, which rapid process, which was also confirmed in research conduct- wasalsoobservedinthestrainsusedinour study [49]. ed by Kumar et al.(2018), as well as in our studies [43, 45]. Similar results were obtained by Franco et al. (2011) and Moreover, tested S. cerevisiae strains varied in their reduction Zou et al. (2012), whereas in a study by García et al. (2018), of AFB concentration (on average 57.92% after 6 h of incu- even though they did not observe DON binding by bation, another 3.07% after 12 h of incubation, with a final Lactobacillus rhamnosus RC007, based on their results, they 3.69% reduction after 24 h), which is consistent with the re- concluded that Lact. rhamnosus contributed to counteract tox- sults obtained by Pizzolitto et al. (2012a) and Poloni et al. ic effect of DON and helped to maintain healthy gastrointes- (2017) [46, 47]. Strain-dependent AFB detoxification, by tinal tract of pigs [50–52]. In our studies, similarly low levels bacteria and yeast, was also observed in the results of studies of activity of S. cerevisiae were demonstrated, decreasing the by Pizzolitto et al. (2011) [48]. The detoxification activity of concentration of DON by 18.01%, on average, after 6 h of yeast strains to reduce the concentration of AFB was similar incubation, by 8.80% after another 6 h and an average of to that of the tested Lactobacillus, and what is also known in 33.03% of the initial concentration after 24 h of incubation. the case of yeast is that the cell wall components are also These results are in line with results obtained by Campagnollo responsible for the binding of the AFB mycotoxin [48]. et al. (2015) [53]. Based on these data, a significantly weaker On the basis of the results obtained, we conclude that DON binding activity by the analysed strains of microorgan- strains of Lactobacillus isolated from the intestinal content isms was also observed in comparison to other mycotoxins, of monogastric animals showed higher binding activity to- namely AF, ZEN or FUM, as also observed by Niderkorn wards DON, than did strains derived from plant silages. et al. (2007) and Campagnollo et al. (2015) [49, 53]. After 6 h of incubation, DON concentration was reduced by The 12 Lactobacillus strains analysed reduced FB and the bacterial strains tested by 5.76–21.64% (mean, 15.70%); FB mixture concentrations by the most significant amounts after 12 h, it was reduced by an average of another 7.39%; and among all mycotoxins tested. The FUM concentrations de- after 24 h, the concentration of DON was reduced by 19.28– creased on average by over 51.23% after 6 h of incubation, 39.37% (mean, 29.55%) of the initial concentration of the and a further 8.61% after 12 h. After 24 h of incubation, the mycotoxin. Niederkorn et al. (2007) showed low concentrations were maintained at an average level of Probiotics & Antimicro. Prot. 29.53 μg/ml (reduction by an average of 70.47% of the initial detoxification by S. cerevisiae strains in their studies and FUM concentration). The greatest reduction in concentration found the yeast’s reduction of the concentration of T-2 toxin occurred after 6 h, while the FUM binding process was ob- was also the result of mycotoxin binding to the yeast cell wall served up to 24 h of incubation, which is comparable to the [60]. results obtained by Abbès et al. (2016) [54]. Zhao et al. (2016) The ability of Lactobacillus strains to bind ZEN was sim- also showed that the process of FUM binding to the surface of ilar to that observed in the case of AFB and the detoxification Lactobacillus is rapid, as they observed the binding after 1 h level of the mycotoxin was mean of 56.90% after 24 h of of incubation [55]. In fact, Pizzolitto et al. (2012b) found that incubation. Our results showed that the greatest reduction in it was an immediate process, occurring after only a minute of ZEN concentrations, by 27.55–58.79% (mean, 42.73%), oc- incubation [56]. Deepthi et al. (2016) also reported high FB curred after 6 h of incubation. The ZEN concentrations were removal (61.7%) by Lact. plantarum MYS6, after 4 h of in- reduced further by an average of 8.17% after the next 6 h of cubation; furthermore, they observed that tested strain sup- incubation, finally reaching an average level of 43.10 μg/ml pressed fumonisin biosynthesis [57]. In our study, high after 24 h (an average 56.90% of the initial concentration of FUM detoxification levels exhibited by Lactobacillus sp., ZEN was bound by the bacterial strains). The capability of which was dependent on the strain tested and mycotoxin type, Lactobacillus strains to decrease ZEN concentration was also coincided with the results obtained by Niderkorn et al. (2007), found by Zhao et al. (2015); ZEN binding activities on the cell Abbès et al. (2016) and Zhao et al. (2016) [49, 54, 55]. walls of the 27 tested Lact. plantarum strains were weaker Moreover, Zhao et al. (2016) also noted that peptidoglycan compared to those of the strains we used in our study [61]. is primarily responsible for the binding of FUM, whereas Furthermore, Zhao et al. established that the binding of ZEN teichoic acid has little effect on mycotoxin binding [55]. On to the cell wall of Lactobacillus is an immediate process, the other hand, Martinez Tuppia et al. (2016), showed that FB which is consistent with the results we obtained [61]. detoxification might not only be a matter of adsorption, but Niderkorn et al. (2006), Čvek et al. (2012), Sangsila et al. also the result of biodegradation to other compounds (e.g. (2016) and Vega et al. (2017) showed comparable results from hydrolysed fumonisin B1, HFB ); however, this was not ob- their studies indicating great potential of Lactobacillus sp. to served for all analysed Lactobacillus sp. strains in their study reduce ZEN concentration [62–65]. We found that [58]. In our research, high and diverse activity of the analysed S. cerevisiae strains reduced ZEN concentration by 24.40– S. cerevisiae strains reducing FUM concentration was also 42.36% (average 34.45%) after 6 h of incubation, with an shown, and the results agree with those of Pizzolitto et al. additional 11.59% after 12 h and an average 5.82% after (2012b) [56]. Furthermore, Armando et al. (2013), who also 24 h. Similar results were obtained by Keller et al. (2015); observed high FB binding properties of S. cerevisiae yeast they concluded that at the beginning, large amounts of ZEN (strain RC016), observed that ability of binding the mycotoxin is bound to the yeast cell wall due to the higher concentration increases along with concentration [59]. Our results also of the mycotoxin in the environment [66]. The ZEN detoxifi- showed that the strains isolated from the feed were cation activity of the bacteria and yeast strains used in this characterised by a slightly lower ability to remove FUM study, was strain-dependent, a result also shown by (67.43% after 24 h of incubation) than achieved by baker’s Armando et al. (2012) and Zhao et al. (2015) [61, 67]. yeast (70.71–74.47% reduction of FUM concentration after 24 h of incubation) or strains isolated from a distillery envi- ronment (72.88% and 73.04% reduction of mycotoxin con- centration after 24 h of incubation). Conclusions The concentrations of T-2 toxin were decreased by the test- ed strains of Lactobacillus and S. cerevisiae to a high degree, Our data allowed us to observe the potential of Lactobacillus where reduction exceeded a mean of 60% after 24 h of incu- and S. cerevisiae to lower the concentrations of the myco- bation. After 6 h of incubation, the Lactobacillus strains re- toxins AFB , DON, FUM, T-2 and ZEN. We found that the duced the concentration of T-2 by 26.68–51.75% (average detoxification of these compounds is a rapid process. To a 42.64%), and after 24 h, it remained at a level of 31.09– great extent, the concentration of the toxins decreases within 50.10 μg/ml (average 39.01 μg/ml). Bacterial strains used in 6 h of incubation, and can last up to 24 h. Nevertheless, the our study showed higher T-2 binding affinity than shown by differences in concentrations were not so significant at a the four strains analysed by Zou et al. (2012) [51]. Only in prolonged period of 24-h incubation. relation to the T-2 toxin did the six S. cerevisiae strains show The Lactobacillus and S. cerevisiae strains analysed significantly stronger detoxification properties (average T-2 showed detoxification properties towards mycotoxins binding after 24 h of incubation was 69.18%) than the tested (AFB ,DON,FUM,T-2,ZEN)inthisinvitrostudy. bacterial strains (average decontamination rate after 24 h was Therefore, these strains, after further investigation, could be 61.45%). Zou et al. (2015) obtained a similar level of T-2 used as food and feed additives to detoxify contaminating Probiotics & Antimicro. Prot. 8. Ji C, Fan Y, Zhao L (2016) Review on biological degradation of mycotoxins, which pose potential threats to human and animal mycotoxins. Animal Nutr 2(3):127–133. https://doi.org/10.1016/j. health. aninu.2016.07.003 9. Zacharisova M, Dzuman Z, Veprikova Z, Hajkova K, Jiru M, Acknowledgements We would like to thank the National Centre for Vaclavikova M, Zacharisaova A, Pospichalova M, Florian M, Research and Development for the financial support of publication of this Hajslova J (2014) Occurrence of multiple mycotoxins in paper within the project PBS3/A8/32/2015 realised within the framework European feedingstuffs, assessment of dietary intake by farm ani- of the Program of Applied Studies. mals. Anim Feed Sci Technol 193:124–140. https://doi.org/10. 1016/j.anifeedsci.2014.02.007 10. Neme K, Mohammed A (2017) Mycotoxin occurrence in grains Funding This research was funded by the National Centre for Research and the role of postharvest management as a mitigation strategies. and Development within the project PBS3/A8/32/2015. A review. Food Control 78:412–425. https://doi.org/10.1016/j. foodcont.2017.03.012 Compliance with Ethical Standards 11. Kosicki R, Błajet-Kosicka A, Grajewski J, Twarużek M (2016) Multiannual mycotoxin survey in feed materials and feedingstuffs. Conflict of Interest The authors declare that they have no conflict of Anim Feed Sci Technol 215:165–180. https://doi.org/10.1016/j. interest. anifeedsci.2016.03.012 12. European Parliament and the Council (2002) Directive 2002/32/EC of the European Parliament and of the Council of 7 May 2002 on Ethical Approval This article does not contain any studies with human undesirable substances in animal feed - Council statement. https:// participants or animals performed by any of the authors. eur-lex.europa.eu/resource.html?uri=cellar:aca28b8c-bf9d-444f- Open Access This article is distributed under the terms of the Creative b470-268f71df28fb.0004.02/DOC_1&format=PDF. Accessed 06 Commons Attribution 4.0 International License (http:// April 2018 creativecommons.org/licenses/by/4.0/), which permits unrestricted use, 13. The Commission Of The European Communities (2006) distribution, and reproduction in any medium, provided you give appro- Commission Regulation (EC) No 1881/2006 of 19 December priate credit to the original author(s) and the source, provide a link to the 2006 setting maximum levels for certain contaminants in food- Creative Commons license, and indicate if changes were made. stuffs. http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri= CELEX:32006R1881&from=en. Accessed 06 April 2018 14. The Commission Of The European Communities (2006) Commission recommendation 2006/576/EC of 17 August 2006 Publisher’sNote Springer Nature remains neutral with regard to jurisdic- on the presence of deoxynivalenol, zearalenone, ochratoxin A, T- tional claims in published maps and institutional affiliations. 2 and HT-2 and fumonisins in products intended for animal feed- ing. http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri= CELEX:32006H0576&from=EN. Accessed 06 April 2018 15. Medina A, Magan N (2011) Temperature and water activity effects on production of T-2 and HT-2 by Fusarium langsethiae strains References from north European countries. Food Microbiol 28:392–398. ht tps://doi.org/10.1016/j.fm.2010.09.012 1. Smith MC, Madec S, Coton E, Hymery N (2016) Natural co- 16. Milani JM (2013) Ecological conditions affecting mycotoxin pro- occurrence of mycotoxins in foods and feeds and their in vitro duction in cereals: a review. Vet Med (Praha) 58(8):405–411. combined toxicological effects. Toxins 8(4):94. https://doi.org/10. https://doi.org/10.17221/6979-VETMED 3390/toxins8040094 17. The Commission Of The European Communities (2013) 2. Alshannaq A, Yu JH (2017) Occurrence, toxicity, and analysis of Commission Recommendation 2013/165/EU of 27 March 2013 major mycotoxins in food. Int J Environ Res Public Health 14(6): on the presence of t-2 and ht-2 toxin in cereals and cereal products. 632. https://doi.org/10.3390/ijerph14060632 http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri= 3. Liew WPP, Mohd-Redzwan S (2018) Mycotoxin: its impact on gut CELEX:32013H0165&from=EN. Accessed 06 April 2018 health and microbiota. Front Cell Infect Microbiol 8(60). https:// 18. Nugmanov A, Beishova I, Kokanov S, Lozowicka B, Kaczynski P, doi.org/10.3389/fcimb.2018.00060 Konecki R, Snarska K, Wołejko E, Sarsembayeva N, Abdigaliyeva 4. Gallo A, Giuberti G, Frisvad JC, Bertuzzi T, Nielsen KF (2015) T (2018) Systems to reduce mycotoxin contamination of cereals in Review on mycotoxin issues in ruminants: occurrence in forages, the agricultural region of Poland and Kazakhstan. Crop Prot 106: effects of mycotoxin ingestion on health status and animal perfor- 64–71. https://doi.org/10.1016/j.cropro.2017.12.014 mance and practical strategies to counteract their negative effects. 19. The European Parliament And The Council Of The European Toxins 7(8):3057–3111. https://doi.org/10.3390/toxins7083057 Union (2017) Regulation (EU) 2017/625 of the European 5. Pinotti L, Ottoboni M, Giromini C, Dell’Orto V, Cheli F (2016) Parliament and of the Council of 15 March 2017 on official controls Mycotoxin contamination in the EU feed supply chain: a focus on and other official activities performed to ensure the application of cereal byproducts. Toxins 8(2):45. https://doi.org/10.3390/ food and feed law, rules on animal health and welfare, plant health toxins8020045 and plant protection products. https://eur-lex.europa.eu/legal- 6. Freire L, Sant’Ana AS (2018) Modified mycotoxins: an updated content/EN/TXT/PDF/?uri=CELEX:32017R0625&from=EN. review on their formation, detection, occurrence, and toxic effects. Accessed 06 April 2018 Food Chem Toxicol 111:189–205. https://doi.org/10.1016/j.fct. 20. Aiko V, Mehta L (2015) Occurrence, detection and detoxification of 2017.11.021 mycotoxins. J Biosci 45(5):943–954. https://doi.org/10.1007/ s12038-015-9569-6 7. Streit E, Schatzmayr G, Tassis P, Tzika E, Marin D, Taranu I, Tabacu C, Nicolau A, Aprodu I, Puel O, Oswald IP (2012) 21. Loi M, Fanelli F, Liuzzi VC, Logrieco AF, Mulè G (2017) Current situation of mycotoxin contamination and co-occurrence Mycotoxin biotransformation by native and commercial enzymes: present and future perspectives. Toxins 9(4):111. https://doi.org/10. in animal feed-focus on Europe. Toxins 4(10):788–809. https:// doi.org/10.3390/toxins4100788 3390/toxins9040111 Probiotics & Antimicro. Prot. 22. Zhu Y, Hassan YI, Lepp D, Shao S, Zhou T (2017) Strategies and 40. Liew WPP, Nurul-Adilah Z, Than LTL, Mohd-Redzwan S (2018) methodologies for developing microbial detoxification systems to The binding efficiency and interaction of Lactobacillus casei mitigate mycotoxins. Toxins 9(4):130. https://doi.org/10.3390/ Shirota toward aflatoxin B . Front Microbiol 9(1503). https://doi. toxins9040130 org/10.3389/fmicb.2018.01503 41. Hernandez-Mendoza A, Garcia HS, Steele JL (2009) Screening of 23. Juodeikiene G, Bartkiene E, Cernauskas D, Cizeikiene D, Zadeike Lactobacillus casei strains for their ability to bind aflatoxin B . D, Lele V, Bartkevics V (2018) Antifungal activity of lactic acid Food Chem Toxicol 47:1064–1068. https://doi.org/10.1016/j.fct. bacteria and their application for Fusarium mycotoxin reduction in 2009.01.042 malting wheat grains. LWT-Food Sci Technol 89:307–314. https:// doi.org/10.1016/j.lwt.2017.10.061 42. Huang L, Duan C, Zhao Y, Gao L, Niu C, Xu J, Li S (2017) Reduction of aflatoxin b toxicity by lactobacillus plantarum c88: 24. El-Nezami H, Kankaanpaa P, Salminen S, Ahokas J (1998) Ability 1 a potential probiotic strain isolated from Chinese traditional of dairy strains of lactic acid bacteria to bind a common food car- fermented food Btofu^. PLoS One 12(1):e0170109. https://doi. cinogen, aflatoxin B1. Food Chem Toxicol 36:321–326 org/10.1371/journal.pone.0170109 25. FAO/WHO (2002) Guidelines for the evaluation of probiotics in 43. Kumar SS, Bashisht A, Venkateswaran G, Hariprasad P, Gayathri D food. Food and Agriculture Organization of the United Nations (2018) Characterization of novel Lactobacillus fermentum from and World Health Organization Working Group Report. http:// curd samples of indigenous cows from Malnad region, Karnataka, www.fao.org/3/a-a0512e.pdf. Accessed 24 March 2018 for their aflatoxin B binding and probiotic properties. Probiotics 26. Vinderola G, Ritieni A (2015) Role of probiotics against myco- Antimicrob Proteins. https://doi.org/10.1007/s12602-018-9479-7 toxins and their deleterious effects. J Food Res 4(1):10–21. 44. Hernandez-Mendoza A, Guzman-de-Peña D, Garcia HS (2009) https://doi.org/10.5539/jfr.v4n1p10 Key role of 383 teichoic acids on aflatoxin B 384 binding by 27. Śliżewska K, Chlebicz A (2017) Strains of lactic acid bacteria from probiotic bacteria. J Appl Microbiol 107:395–403. https://doi.org/ the genus Lactobacillus. Patent Application no 422603 10.1111/j.1365-3852672.2009.04217.x 28. Aflatoxin B from Aspergillus flavus, Sigma-Aldrich. https://www. 45. Gratz S, Mykkänen H, El-Nezami H (2005) Aflatoxin B binding sigmaaldrich.com/catalog/product/sigma/a6636?lang=pl&region= by a mixture of Lactobacillus and Propionibacterium: in vitro ver- PL. Accessed 21 March 2018 sus ex vivo. J Food Prot 68(11):2470–2474. https://doi.org/10.4315/ 29. Deoxynivalenol, Sigma-Aldrich. https://www.sigmaaldrich.com/ 0362-028X-68.11.2470 catalog/product/sial/32943?lang=pl&region=PL. Accessed 21 46. Pizzolitto RP, Armando MR, Combina M, Cavaglieri LR, Dalcero March 2018 AM, Salvano MA (2012) Evaluation of Saccharomyces cerevisiae 30. Fumonisin B , Sigma-Aldrich. https://www.sigmaaldrich.com/ strains as probiotic agent with aflatoxin B adsorption ability for use catalog/product/sial/34139?lang=pl&region=PL&cm_sp=Insite-_- in poultry feedstuffs. J Environ Sci Health B 47(10):933–941. prodRecCold_xorders-_-prodRecCold2-1. Accessed 21 https://doi.org/10.1080/03601234.2012.706558 March 2018 47. Poloni V, Salvato L, Pereyra L, Oliveira A, Rosa C, Cavaglieri L, 31. Fumonisin B , Sigma-Aldrich. https://www.sigmaaldrich.com/ Keller KM (2017) Bakery by-products based feeds borne- catalog/product/sial/34142?lang=pl&region=PL&cm_sp=Insite-_- Saccharomyces cerevisiae strains with probiotic and antimycotoxin prodRecCold_xorders-_-prodRecCold2-2. Accessed 21 effects plus antibiotic resistance properties for use in animal pro- March 2018 duction. Food Chem Toxicol 107:630–636. https://doi.org/10.1016/ 32. T-2 toxin, Sigma-Aldrich. https://www.sigmaaldrich.com/catalog/ j.fct.2017.02.040 product/sigma/t4887?lang=pl&region=PL. Accessed 21 48. Pizzolitto RP, Bueno DJ, Armando MR, Cavaglieri L, Dalcero AM, March 2018 Salvano MA (2011) Binding of aflatoxin B to lactic acid bacteria 33. Zearalenone, Sigma-Aldrich. https://www.sigmaaldrich.com/ and Saccharomyces cerevisiae in vitro: a useful model to determine catalog/product/sigma/z2125?lang=pl&region=PL. Accessed 21 the most efficient microorganism. In: Guevara-Gonzalez RG (ed) March 2018 Aflatoxins. Biochemistry and molecular biology, InTech. https:// 34. Devreese M, De Backer P, Croubels S (2013) Different methods to doi.org/10.5772/23717. https://www.intechopen.com/books/ counteract mycotoxin production and its impact on animal health. aflatoxins-biochemistry-and-molecular-biology/binding-of- Vlaams Diergeneeskd Tijdschr 82(4):181–190 aflatoxin-b1-to-lactic-acid-bacteria-and-saccharomyces-cerevisiae- 35. Karlovsky P, Suman M, Berthiller F, De Meester J, Eisenbrand G, in-vitro-a-useful-model. Accessed 28 March 2018 Perrin I, Oswald IPm Speijers G, Chiodini A, Recker T, Dussort P 49. Niderkorn V, Morgavi DP, Pujos E, Tissandier A, Boudra H (2007) (2016) Impact of food processing and detoxification treatments on Screening of fermentative bacteria for their ability to bind and mycotoxin contamination. Mycotoxin Res 32(4):179–205. https:// biotransform deoxynivalenol, zearalenone and fumonisins in an doi.org/10.1007/s12550-016-0257-7 in vitro simulated corn silage model. Food Addit Contam 24(4): 36. Moslehi-Jenabian S, Pedersen LL, Jespesen L (2010) Beneficial 406–415. https://doi.org/10.1080/02652030601101110 effects of probiotic and food borne yeasts on human health. 50. Franco TS, Garcia S, Hirooka EY, Ono YS, Santos JS (2011) Lactic Nutrients 2(4):449–473. https://doi.org/10.3390/nu2040449 acid bacteria in the inhibition of Fusarium graminearum and 37. Śliżewska K, Smulikowska S (2011) Detoxification of aflatoxin B deoxynivalenol detoxification. J Appl Microbiol 111:739–748. and change in microflora pattern by probiotic in vitro fermentation https://doi.org/10.1111/j.1365-2672.2011.05074.x of broiler feed. Anim Feed Sci Technol 20:300–309. https://doi.org/ 51. Zou ZY, He ZF, Li HJ, Han PF, Meng X, Zhang Y, Zhou F, Ouyang 10.22358/jafs/66187/2011 KP, Chen XY, Tang J (2012) In vitro removal of deoxynivalenol and 38. Vila-Donat P, Marin S, Sachis V, Ramos AJ (2018) A review of the T-2 toxin by lactic acid bacteria. Food Sci Biotechnol 21(6):1677– mycotoxin adsorbing agents, with an emphasis on their multi- 1683. https://doi.org/10.1007/s10068-012-0223-x binding capacity, for animal feed decontamination. Anim Feed 52. García GR, Payros D, Pinton P, Dogi CA, Laffitte J, Neves M, Sci Technol 114:246–259. https://doi.org/10.1016/j.fct.2018.02. González Pereyra ML, Cavaglieri LR, Oswald IP (2018) 044 Intestinal toxicity of deoxynivalenol is limited by Lactobacillus rhamnosus RC007 in pig jejunum explants. Arch Toxicol 92: 39. Haskard CA, El-Nezmi HS, Kankaanpää PE, Salminen S, Ahokas 983–993. https://doi.org/10.1007/s00204-017-2083-x JT (2001) Surface binding of aflatoxin B by lactic acid bacteria. 53. Campagnollo FB, Franco LT, Rottinghaus GE, Kobashigawa E, Appl Environ Microbiol 67(7):3086–3091. https://doi.org/10.1128/ Ledoux DR, Daković A, Oliveira CAF (2015) In vitro evaluation AEM.67.7.3086-3091.2001 Probiotics & Antimicro. Prot. of the ability of beer fermentation residue containing 60. Zou Z, Sun J, Huang F, Feng Z, Li M, Shi R, Ding J, Li H (2015) Saccharomyces cerevisiae to bind mycotoxins. Food Res Int 77: In vitro removal of T-2 toxin by yeasts. J Food Safety 35:544–550. 643–648. https://doi.org/10.1016/j.foodres.2015.08.032 https://doi.org/10.1111/jfs.12204 61. Zhao L, Jin H, Zhang R, Ren H, Zhang X, Yu G (2015) 54. Abbès S, Salah-Abbès JB, Jebali R, Younes RB, Oueslati R (2016) Detoxification of zearalenone by three strains of Lactobacillus Interaction of aflatoxin B and fumonisin B in mice causes 1 1 plantarum from fermented food in vitro. Food Control 54:158– immunotoxicity and oxidative stress: possible protective role using 164. https://doi.org/10.1016/j.foodcont.2015.02.003 lactic acid bacteria. J Immunotoxicol 13(1):46–54. https://doi.org/ 62. Niderkorn V, Boudra H, Morgavi DP (2006) Binding of Fusarium 10.3109/1547691X.2014.997905 mycotoxins by fermentative bacteria in vitro. J Appl Microbiol 101: 55. Zhao H, Wang X, Zhang J, Zhang J, Zhang B (2016) The mecha- 849–856. https://doi.org/10.1111/j.1365-2672.2006.02958.x nism of Lactobacillus strains for their ability to remove fumonisins 63. Čvek D, Markov K, Frece J, Friganović M, Duraković L, Delaš F B and B . Food Chem Toxicol 97:40–46. https://doi.org/10.1016/j. 1 2 (2012) Adhesion of zearalenone to the surface of lactic acid bacteria fct.2016.08.028 cells. Croat J Food Tech Biotech Nutr 7:49–52 56. Pizzolitto RP, Salvano MA, Dalcero AM (2012) Analysis of 64. Sangsila A, Faucet-Marquis V, Pfohl-Leszkowicz A, Itsaranuwat P fumonisin B removal by microorganisms in co-occurrence with (2016) Detoxification of zearalenone by Lactobacillus pentosus aflatoxin B and the nature of the binding process. Int J Food strains. Food Control 62:187–192. https://doi.org/10.1016/j. Microbiol 156:214–221. https://doi.org/10.1016/j.ijfoodmicro. foodcont.2015.10.031 2012.03.024 65. Vega MF, Dieguez SN, Riccio B, Aranguren S, Giordano A, 57. Deepthi BV, Poornachandra Rao K, Chennapa G, Naik MK, Denzoin L, Soraci AL, Tapia MO, Ross R, Apás A, González SN Chandrashekara KT, Sreenivasa MY (2016) Antifungal attributes (2017) Zearalenone adsorption capacity of lactic acid bacteria iso- of Lactobacillus plantarum MYS6 against fumonisin producing lated from pigs. Braz J Microbiol 48(4):715–723. https://doi.org/ Fusarium proliferatum associated with poultry feeds. PLoS One 10.1016/j.bjm.2017.05.001 11(6):e0155122. https://doi.org/10.1371/journal.pone.0155122 66. Keller L, Abrunhosa L, Keller K, Rosa CA, Cavaglieri L, Venâncio 58. Martinez Tuppia C, Atanasova-Penichon V, Chéreau S, Ferrer N, A (2015) Zearalenone and its derivatives α-zearalenol and β- Marchegay G, Savoie J, Richard-Forget F (2017) Yeast and bacteria zearalenol decontamination by Saccharomyces cerevisiae strains from ensiled high moisture maize grains as potential mitigation isolated from bovine forage. Toxins 7(8):3297–3308. https://doi. agents of fumonisin B . J Sci Food Agric 97:2443–2452. https:// org/10.3390/toxins7083297 doi.org/10.1002/jsfa.8058 67. Armando MR, Pizzolitto RP, Dogi CA, Cristofolini A, Merkis C, 59. Armando M, Galvagno M, Dogi C, Cerrutti P, Dalcero A, Poloni V, Dalcero AM, Cavaglieri LR (2012) Adsorption of ochra- Cavaglieri L (2013) Statistical optimization of culture conditions toxin A and zearalenone by potential probiotic Saccharomyces for biomass production of probiotic gut-borne Saccharomyces cerevisiae strains and its relations with cell wall thickness. J Appl cerevisiae strain able to reduce fumonisin B . J Appl Microbiol Microbiol 113:256–264. https://doi.org/10.1111/j.1365-2672.2012. 114:1338–1346. https://doi.org/10.1111/jam.12144 05331.x http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Probiotics and Antimicrobial Proteins Springer Journals

In Vitro Detoxification of Aflatoxin B1, Deoxynivalenol, Fumonisins, T-2 Toxin and Zearalenone by Probiotic Bacteria from Genus Lactobacillus and Saccharomyces cerevisiae Yeast

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Chemistry; Chemistry/Food Science, general; Applied Microbiology; Microbiology; Protein Science; Nutrition
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1867-1306
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10.1007/s12602-018-9512-x
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Abstract

The aim of the following research was to determine the detoxification properties of probiotic Lactobacillus sp. bacteria (12 strains) and S. cerevisiae yeast (6 strains) towards mycotoxins, such as aflatoxin B , deoxynivalenol, fumonisins, T-2 toxin and zearalenone, which pose as frequent feed contamination. The experiment involved analysing changes in concentration of mycotoxins in PBS solutions, after 6, 12 and 24 h of incubation with monocultures of tested microorganisms, measured by high-performance liquid chromatography (HPLC). We found that all strains detoxified the mycotoxins, with the highest reduction in concentration observed for the fumonisin B and B mixture, ranging between 62 and 77% for bacterial strains and 67–74% for 1 2 yeast. By contrast, deoxynivalenol was the most resistant mycotoxin: its concentration was reduced by 19–39% by Lactobacillus sp. strains and 22–43% by yeast after 24 h of incubation. High detoxification rates for aflatoxin B , T-2 toxin and zearalenone were also observed, with concentration reduced on average by 60%, 61% and 57% by Lactobacillus,respectively, and65%, 69% and 52% by yeast, respectively. The greatest extent of reduction in the concentration for all mycotoxins was observed after 6 h of incubation; however, a decrease in concentration was noted even after 24 h of incubation. Thus, the tested microorganisms can potentially be used as additives to decrease the concentrations of toxins in animal feed. . . . . Keywords Mycotoxins Detoxification Probiotics Lactobacillus Saccharomyces cerevisiae Introduction fumonisins (FUM), along with zearalenone (ZEN) are the most prevalent mycotoxin-related contamination found in Mycotoxins are secondary metabolites with low molecular fodder [5, 6]. AFB is most frequently found in feed in the mass (~ 700 Da), which are synthesised by filamentous fungi, European Union (> 98% of tested samples); however, DON belongingmostlytothe Ascomycota phylum. The most com- (~ 90% of tested samples) and ZEN (~ 70% of tested samples) mon source of food and feed contamination are mycotoxins are often detected as well. The presence of FUM and OTA, on produced by the fungi Aspergillus, Penicillium and Fusarium the other hand, are more sparsely observed [7]. genera [1–3]. Other mycotoxin-producing fungi include Plants are contaminated with mycotoxins, synthesised by Alternaria, Chaetomium, Cladosporium, Claviceps, filamentous fungi, most frequently at the time of cultivation in Diplodia, Myrothecium, Monascus, Phoma, Phomopsis, the fields (e.g. mycotoxin produced mostly by Fusarium sp.). Pithomyces, Trichoderma and Stachybotrys [4]. Aflatoxins Likewise, under favourable growth conditions of temperature (AF), ochratoxin (OT), trichotecens, including and humidity, mycotoxin-producing fungi, such as deoxynivalenol (DON) and T-2 toxin (T-2), as well as Aspergillus sp. and Penicillium sp., are also found in food and feed that are stored [2, 8, 9]. In stored grains, moisture content within 16–30%, high temperature reaching 25–30 °C, * Agnieszka Chlebicz and high relative air humidity (80–100%) are conditions that agnieszka.chlebicz@edu.p.lodz.pl stimulate growth of filamentous fungi and mycotoxin produc- * Katarzyna Śliżewska tion [10]. The concentration of toxins in input materials (e.g. katarzyna.slizewska@p.lodz.pl corn, grass, clover) is not reduced to a sufficient degree while they are being processed into feed, as these metabolites are Institute of Fermentation Technology and Microbiology, Department resistant to high and low temperatures, even after long storage of Biotechnology and Food Sciences, Lodz University of Technology, Wólczańska 171/173, 90-924 Łódź, Poland period [9, 11]. Therefore, these toxins constitute a threat, as Probiotics & Antimicro. Prot. they can enter the human food chain through products such as mycotoxins to less toxic metabolites, and the proper treatment milk, meat or eggs [2]. Furthermore, humans are exposed to of food or feed by fermentation are also used for biological mycotoxin-related intoxications while consuming foods of detoxification [9, 20–23]. In comparison to physical and plant origin, for instance hazelnuts, almonds, grains and fruits chemical methods, biological detoxification is more efficient, [8]. Therefore, European Union legislation specifies tolerable specific and safer for the environment [22]. daily intake (TDI) for a variety of mycotoxins, in addition to In 1998, El-Nazami performed pioneering in vitro studies providing guidance values for their concentrations in animal on the binding properties of mycotoxin by lactic acid bacteria, feedstuffs (Table 1)[18]. which has initiated a systematic search for microorganisms The regulation (EU) 2017/625 of the European Parliament having specific abilities to adsorb mycotoxins [24]. Among and of the Council of 15 March 2017, regarding official con- other microbes identified for biological detoxification pur- trols and other official activities performed to ensure the ap- poses, probiotic microorganisms, defined by FAO/WHO plication of food and feed law, rules on animal health and (2002) as ‘live microorganisms, which when administered in welfare, plant health and plant protection products will come adequate amounts confer a health benefit on the host’,were into effect on 14th December 2019. According to this regula- identified as microbes that bind and adsorb mycotoxins [25]. tion, Member States are obliged to establish multiannual plans Probiotic microorganisms, such as bacteria belonging to gen- and carry out food and feed controls, to ensure safety in the era Lactobacillus, Bifidobacterium,as well as Enterococcus agri-food chain, as well as animal welfare and health, thereby faecium,andtheyeasts Saccharomyces cerevisiae and providing safe food [19]. Saccharomyces boulardii have been shown to have mycotox- The process of detoxification or mycotoxin removal is in detoxification properties [26]. complicated, especially because of heat stability of these com- The aim of our study was to determine detoxification prop- pounds, and their breakdown into toxic products. erties of probiotic strains of Lactobacillus sp. and S. cerevisiae Nevertheless, detoxification may be accomplished by applica- towards mycotoxins, which often contaminate feed for live- tion of the following methods: physical (cooking, baking, mi- stock animals. This study was part of one of the stages of crowave heating, radiation, etc.), chemical (use of ammonia, strain selection for designing synbiotic preparations for poul- hydrochloric acid, salicylic, sulfamide, sulfosalicylic, try and swine. anthranilic, benzoic, boric, oxalic or propionic acid), and bio- logical [20]. For biological detoxification, plant extracts, such as piperine, lutein, carotenoids or essential oils, as well as Materials and Methods enzymes, such as AF decomposing peroxidase or laccase, and FUM degradative carboxylesterase or aminotransferase Biological Materials are used. Microorganisms that are capable of degrading The biological material included potential probiotic bacteria of Lactobacillus genus and strains of the yeast S. cerevisiae, Table 1 Optimal conditions for mycotoxins production, TDI in food which are deposited in the Łódź Collection of Pure Cultures products and guidance value in feedstuff in European Union [12–17] 105 of Institute of Fermentation Technology and Mycotoxin TDI in Guidance Optimum Optimum Microbiology at Technical University of Łódź (Table 2). food value in temperature water activity Five Lactobacillus (rhamnosus ŁOCK 1087, paracasei (μg/kg)* feedstuff (12% for mycotoxin for ŁOCK 1091, reuteri ŁOCK 1092, plantarum ŁOCK 0860, moisture) production mycotoxin pentosus ŁOCK 1094) and one S. cerevisiae (ŁOCK 0119) (mg/kg)** (°C) production strain have been documented in patent application no. 422603 AF*** 0.025–15.0 0.005–0.05 33 0.99 [27]. DON 200–1750 0.9–12 26–30 0.995 The detoxification activity of bacteria and yeast was deter- FUM**** 200–2000 5–60 15–30 0.9–0.995 mined against five mycotoxins, namely aflatoxin B1, OTA 0.50–10.0 0.05–0.25 25–30 0.98 fumonisin mixture of fumonisin B1 and B2 (FUM), T-2 toxin, T-2/HT-2 15–1000 0.25–220–30 0.98–0.995 zearalenone and deoxynivalenol (Table 3). Mycotoxins were ZEA 20–200 0.1–325 0.96 suspended in PBS buffer (Calbiochem®, Germany), and so- lutions of 100 μg/ml were prepared. *Depending on a product (e.g. nuts intended for processing or direct consumption, raw grains, milk, dried fruits, spices, infants’ formulas, Bacterial and Yeast Strain Cultivation and Sample processed foods based on cereals, wine, coffee, etc.) Preparation **With a distinction between feed materials and complementary and complete feed mixtures ***Depending on form Lactobacillus sp. were cultivated in de Man, Rogosa, and ****Sum of FB and FB Sharpe (MRS) broth (Merck, Germany) at 37 °C, while 1 2 Probiotics & Antimicro. Prot. Table 2 Strains whose Microorganism Collection number Source of isolation mycotoxin detoxification properties has been studied Bacteria Lact. brevis ŁOCK 1093 Plant silages Lact. casei ŁOCK 0911 Milk fermented beverages Lact. casei ŁOCK 0915 Milk fermented beverages Lact. paracasei ŁOCK 1091 Caecal content of sow Lact. pentosus ŁOCK 1094 Broiler chicken dung Lact. plantarum ŁOCK 0860 Plant silages Lact. plantarum ŁOCK 0862 Plant silages Lact. reuteri ŁOCK 1092 Piglet caecal content Lact. reuteri ŁOCK 1096 Winear pig’s intestinal content Lact. rhamnosus ŁOCK 1087 Turkey dung Lact. rhamnosus ŁOCK 1088 Broiler chicken’s intestinal content Lact. rhamnosus ŁOCK 1089 Broiler chicken’s intestinal content Yeast S. cerevisiae ŁOCK 0068 Forage S. cerevisiae ŁOCK 0113 Distillers’ yeast, potato with grain S. cerevisiae ŁOCK 0119 Distillers’ yeast, grain S. cerevisiae ŁOCK 0137 Baker’syeast S. cerevisiae ŁOCK 0140 Baker’syeast S. cerevisiae ŁOCK 0142 Baker’syeast S. cerevisiae was grown at 30 °C, in yeast extract–peptone– was used with a 250 × 4.6 mm size ACE 5 C18 column (Advanced glucose broth (YPG Broth, Merck, Germany). Both bacteria Chromatography Technologies (ACT), Scotland). Mycotoxins and yeast cultures were grown in normal oxygen conditions were identified by comparing the retention times of the peak with for 24 h. After 24 h of incubation, monocultures of the standard solutions. The mycotoxin concentrations were deter- analysed strains, in three repetitions, were centrifuged at rela- mined by correlation of peak area of the samples with the stan- tive centrifugal force (RCF) 3468×g for 10 min (Centrifuge dard curves, obtained by HPLC analysis of standard solutions. MPW-251; MPW, Poland). Subsequently, the supernatants were removed and the bacteria and yeast biomass were Statistical Analysis washed three times with PBS buffer to remove any residual culture medium. The cell pellets were again centrifuged under The results presented here constitute the arithmetic mean of the same conditions. Ten millilitres of mycotoxin solutions values from three repetitions, with standard deviation. All sta- were added to the prepared samples, with a defined concen- tistical analyses were carried out using the one-way ANOVA tration of 100 μg/ml for each mycotoxin. These samples were test, with a significance level of p < 0.05 (Origin 6.1 program, further incubated in normal oxygen conditions for 24 h at OriginLab). A comparative Duncan test was carried out at a 37 °C or 30 °C for lactic acid bacteria and yeast, respectively. significance level of p > 0.05 (STATISTICA 10, StatSoft). After 6, 12 and 24 h of incubation, 2 ml of each sample was collected, centrifuged at RCF of 3468×g for 10 min, and the supernatants were filtered with PTFE syringe filters with Results 0.22-μm-diameter pores (Millex-GS, Millipore, USA). As a positive control sample, a solution of analysed mycotoxin in AFB Microbial Detoxification PBS was used, and bacterial or yeast suspension served as negative control sample. Bacteria belonging to the Lactobacillus genus were characterised by their diverse ability to detoxify aflatoxin HPLC Analysis B . In merely after 6 h of incubation, a statistically significant reduction of AFB concentration was noticed, ranging from The prepared samples were subjected to high-performance liquid 35.33 to 79.65 μg/ml (20–65% reduction, 49% on average) chromatography (HPLC) analysis, the parameters of which are compared to the initial mycotoxin concentration of 100 μg/ml. presented in Table 4. Analysis was performed as previously de- In subsequent hours, further reduction in AFB concentration scribed by El-Nazami et al. [24], with modifications. For this pur- was observed. After 24 h of incubation, the concentration of pose, Surveyor liquid chromatography (Thermo Scientific, USA) mycotoxin in the samples was 28.96–55.80 μg/ml (mean, Probiotics & Antimicro. Prot. Table 3 Mycotoxins, that were detoxified by selected strains of potentially probiotic microorganisms (Sigma-Aldrich, available online on https://www. sigmaaldrich.com/, accessed on 21 March 2018 [28–33]) Mycotoxin Chemical structure Producer, catalogue number Producer, catalogue number of HPLC sample AFB Sigma, A6636 Supelco, 46323-u DON Sigma, 32943 Supelco, CRM46911 FUM mixture Sigma, 34143 Fumonisin B – Sigma, 34139 Fumonisin B Fumonisin B – Sigma, 34142 Fumonisin B T-2 Sigma, T4887 Sigma, 34071 ZEA Sigma, Z2125 Supelco, CRM46916 Probiotics & Antimicro. Prot. Table 4 HPLC analysis parameters Parameter Mycotoxins AFB DON FUM T-2 ZEN Column heating – 30 –– – Mobile phase Water/acetonitrile/methanol Water/acetonitrile Gradient methanol/water Methanol/water Methanol/water (60:30:10) (90:10) (70:30 and 80:20) (60:40) (70:30) Fluorescent detector λ (nm) (ex- 360 and 420 – 490 and 450 381 and 470 280 and 460 citation and emission) UV detector λ (nm) – 218 –– – Flow (ml/min) 1 1 1 1 1 40.43 μg/ml). Therefore, there was a reduction of between 44 DON Microbial Detoxification and 71% (mean, 60%) compared to the initial concentration of the mycotoxin. Deoxynivalenol (DON) concentrations were significantly re- S. cerevisiae showed detoxification activity similar to that duced after 6 h of incubation in the presence of the bacteria of analysed strains of Lactobacillus sp. After 6hof incuba- monoculture, varying between 78.39 and 94.24 μg/ml (mean, tion, AFB concentrations were statistically significantly re- 84.30 μg/ml), indicating that these bacteria have the ability to duced by 47–66% (average 58%) and ranged from 33.64 to decrease DON concentration by an average of 16%. Further 53.27 μg/ml. In subsequent hours of incubation, the concen- decrease in DON concentration in the subsequent incubation tration of the mycotoxin further decreased, and after 24 h, the hours was also observed. After 24 h, DON concentrations mycotoxin concentrations were 32.48–4.45 μg/ml (average ranged between 60.63 and 80.72 μg/ml (mean, 70.45 μg/ml), reduction of AFB1 by 65%) (Table 5). thereby showing a reduction of 19–39% (mean, 30%). Table 5 Reduction of AFB concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B B C Lactobacillus brevis ŁOCK 1093 100 39.56 ± 0.99 (60) 37.13 ± 2.04 (63) 32.70 ± 0.59 (67) B B C casei ŁOCK 0911 51.30 ± 2.04 (49) 49.92 ± 1.36 (50) 45.13 ± 0.38 (55) B C D casei ŁOCK 0915 67.18 ± 0.63 (33) 58.98 ± 1.27 (41) 55.80 ± 1.51 (44) B C D paracasei ŁOCK 1091 57.44 ± 1.61 (43) 48.23 ± 1.27 (52) 42.21 ± 1.52 (58) B B B pentosus ŁOCK 1094 44.70 ± 0.41 (55) 38.73 ± 1.03 (61) 28.96 ± 0.58 (71) B B C plantarum ŁOCK 0860 47.93 ± 1.01 (52) 45.88 ± 0.88 (54) 40.62 ± 0.50 (59) B B B plantarum ŁOCK 0862 35.33 ± 0.95 (65) 34.82 ± 0.47 (65) 34.30 ± 1.06 (66) B B B reuteri ŁOCK 1092 45.19 ± 0.97 (55) 44.98 ± 0.60 (55) 43.79 ± 1.81 (56) B C D reuteri ŁOCK 1096 40.79 ± 0.74 (59) 38.41 ± 1.08 (62) 36.02 ± 0.57 (64) B B B rhamnosus ŁOCK 1087 41.42 ± 2.34 (59) 40.32 ± 0.34 (60) 40.19 ± 1.48 (60) B C D rhamnosus ŁOCK 1088 79.65 ± 0.96 (20) 61.04 ± 0.88 (39) 44.83 ± 1.49 (55) B C D rhamnosus ŁOCK 1089 56.24 ± 0.98 (44) 50.43 ± 0.46 (50) 40.61 ± 1.58 (59) Average concentration (μg/ml) (decrease (%)) 50.56 (49) 45.74 (54) 40.43 (60) B B C S. cerevisiae ŁOCK 0068 100 A 46.79 ± 0.62 (53) 44.43 ± 1.35 (56) 38.07 ± 1.35 (62) B C D ŁOCK 0113 53.27 ± 0.69 (47) 48.93 ± 1.75 (51) 41.45 ± 0.63 (59) B C D ŁOCK 0119 42.25 ± 0.73 (58) 34.12 ± 1.05 (66) 31.48 ± 1.05 (69) B B B ŁOCK 0137 34.99 ± 1.85 (65) 34.02 ± 2.30 (66) 32.70 ± 0.79 (67) B B C ŁOCK 0140 41.55 ± 1.62 (58) 39.58 ± 1.42 (60) 35.78 ± 1.39 (64) B B,C C ŁOCK 0142 33.64 ± 0.48 (66) 33.00 ± 1.41 (67) 32.48 ± 0.38 (68) Average concentration (μg/ml) (decrease (%)) 42.08 (58) 39.01 (61) 35.33 (65) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) Probiotics & Antimicro. Prot. S. cerevisiae strains used in the analysis also demonstrated 25.53 and 32.57 μg/ml, demonstrating an average reduction the ability to reduce the concentration of DON in suspension. of 72% of the initial concentration (Table 7). After 6 h, significant reductions of DON concentrations were observed, ranging between 12 and 22% (mean, 18%) of the initial concentration of DON. After a further 6 h, the concen- T-2 Microbial Detoxification trations were 65.57–80.81 μg/ml (mean, 73.20 μg/ml). After 24 h of incubation, the DON concentrations were reduced by After 6 h of incubation, significant reduction of T-2 toxin 22–40% (mean, 33%) relative to the initial concentration of concentrations by monocultures of analysed Lactobacillus mycotoxin (Table 6). sp. strains were observed ranging between 48.25 and 73.32 μg/ml (mean, 57.36 μg/ml). Continued incubation caused a further decrease in T-2 concentration, as a result of FUM Microbial Detoxification which after 24 h of incubation the concentrations of mycotox- in were 31.09–50.10 μg/ml (mean, 39.01 μg/ml). This shift in Bacteria belonging to Lactobacillus genus detoxified the mix- concentration values indicated a reduction of 50–69% (mean, ture of fumonisin B (FB )and B (FB )mycotoxins (FUMs). 1 1 2 2 61%) in relation to the initial quantity of T-2. After 6 h of incubation, the FUM concentration was reduced S. cerevisiae strains subjected to the analysis were by 36–64% (mean, 51%) compared to the initial concentration characterised by diverse T-2 detoxification activity. After 6-h of the mycotoxin mixture. Subsequent incubation resulted in a incubation, a statistically significant decrease in the concen- further significant decrease in FUM concentration, which after tration of T-2 to level of 46.92–54.98 μg/ml (average reduc- 24 h of incubation was reduced by an average of 70% and tion of 49% of initial concentration) was observed. In subse- ranged from 23.08 to 38.42 μg/ml (mean, 29.53 μg/ml). quent hours of incubation, T-2 concentration further de- S. cerevisiae also reduced the concentrations of FUM, sig- creased. The concentration of the mycotoxin after 24 h of nificantly reducing them by 29–60% (mean, 53%) to 40.15– incubation reduced by 60–63% (mean, 61%) of the initial 70.57 μg/ml (mean, 47.19 μg/ml). In subsequent hours of concentration and ranged between 37.36 and 40.40 μg/ml incubation, the concentrations of mycotoxin further declined. (mean, 38.68 μg/ml) (Table 8). Finally, after 24 h, the FUM concentration ranged between Table 6 Reduction of DON concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B C D Lactobacillus brevis ŁOCK 1093 100 94.24 ± 0.77 (6) 87.52 ± 0.95 (12) 80.72 ± 0.33 (19) B C D casei ŁOCK 0911 92.06 ± 1.10 (8) 84.06 ± 1.01 (16) 72.49 ± 0.67 (28) B C D casei ŁOCK 0915 88.73 ± 1.85 (11) 81.93 ± 0.75 (18) 78.06 ± 1.20 (22) B C D paracasei ŁOCK 1091 84.44 ± 1.75 (16) 73.75 ± 3.09 (26) 67.30 ± 1.46 (33) B C D pentosus ŁOCK 1094 78.53 ± 1.19 (21) 72.49 ± 1.88 (27) 66.82 ± 0.65 (33) B C D plantarum ŁOCK 0860 80.34 ± 0.49 (20) 74.12 ± 0.83 (26) 70.25 ± 1.01 (30) B C D plantarum ŁOCK 0862 83.78 ± 0.20 (16) 78.34 ± 1.27 (22) 74.53 ± 1.04 (25) B C D reuteri ŁOCK 1092 79.92 ± 1.02 (20) 69.05 ± 0.44 (31) 60.63 ± 0.59 (39) B C D reuteri ŁOCK 1096 78.36 ± 0.30 (22) 70.58 ± 1.38 (29) 61.31 ± 1.45 (39) B C D rhamnosus ŁOCK 1087 87.85 ± 2.94 (12) 79.04 ± 1.13 (21) 75.05 ± 1.43 (25) B C D rhamnosus ŁOCK 1088 83.04 ± 2.78 (17) 78.14 ± 0.86 (22) 73.74 ± 0.30 (26) B C D rhamnosus ŁOCK 1089 80.35 ± 0.49 (20) 74.01 ± 2.61 (26) 64.55 ± 0.70 (35) Average concentration (μg/ml) (decrease (%)) 84.30 (16) 76.92 (23) 70.45 (30) B C D S. cerevisiae ŁOCK 0068 100 A 77.68 ± 1.53 (22) 70.62 ± 1.17 (29) 64.57 ± 0.57 (35) B C D ŁOCK 0113 88.32 ± 0.88 (12) 80.81 ± 1.45 (19) 78.03 ± 0.82 (22) B C D ŁOCK 0119 80.87 ± 0.85 (19) 65.57 ± 1.52 (34) 57.50 ± 0.83 (43) B B C ŁOCK 0137 84.16 ± 1.40 (16) 80.41 ± 2.27 (20) 76.10 ± 1.32 (24) B C D ŁOCK 0140 80.64 ± 1.71 (19) 71.97 ± 2.04 (20) 63.07 ± 1.49 (37) B C D ŁOCK 0142 80.28 ± 2.07 (20) 69.80 ± 1.61 (30) 60.91 ± 1.24 (39) Average concentration (μg/ml) (decrease (%)) 81.99 (18) 73.20 (27) 66.70 (33) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) Probiotics & Antimicro. Prot. Table 7 Reduction of FUM concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B C D Lactobacillus brevis ŁOCK 1093 100 50.14 ± 1.26 (50) 43.87 ± 1.01 (56) 34.27 ± 1.26 (66) B C C casei ŁOCK 0911 47.80 ± 3.20 (52) 40.63 ± 0.46 (59) 36.32 ± 3.27 (64) B C D casei ŁOCK 0915 48.52 ± 3.21 (51) 41.98 ± 1.81 (58) 33.50 ± 1.11 (67) B C D paracasei ŁOCK 1091 55.98 ± 3.55 (44) 45.56 ± 1.57 (54) 38.42 ± 1.39 (62) B C D pentosus ŁOCK 1094 45.24 ± 0.41 (55) 38.61 ± 1.02 (61) 29.61 ± 1.04 (70) B C D plantarum ŁOCK 0860 35.52 ± 1.92 (64) 30.88 ± 0.69 (70) 25.11 ± 1.67 (75) B C D plantarum ŁOCK 0862 38.72 ± 1.04 (61) 30.54 ± 0.08 (69) 23.19 ± 1.60 (77) B C D reuteri ŁOCK 1092 42.64 ± 1.18 (57) 37.38 ± 1.34 (63) 27.23 ± 1.17 (73) B C D reuteri ŁOCK 1096 41.83 ± 1.20 (58) 34.62 ± 2.32 (65) 28.22 ± 1.05 (72) B C D rhamnosus ŁOCK 1087 56.53 ± 2.13 (43) 45.39 ± 0.36 (55) 24.00 ± 0.69 (76) B C D rhamnosus ŁOCK 1088 63.64 ± 1.98 (36) 53.87 ± 1.27 (46) 31.43 ± 0.95 (69) B C D rhamnosus ŁOCK 1089 58.75 ± 1.35 (41) 38.67 ± 1.08 (61) 23.08 ± 0.08 (77) Average concentration (μg/ml) (decrease (%)) 48.77 (51) 40.17 (60) 29.53 (70) A B C D S. cerevisiae ŁOCK 0068 100 70.57 ± 0.64 (29) 56.03 ± 1.13 (44) 32.57 ± 2.21 (67) B C D ŁOCK 0113 48.73 ± 0.32 (51) 40.12 ± 1.08 (60) 27.12 ± 1.06 (73) B C D ŁOCK 0119 54.98 ± 1.73 (45) 43.91 ± 1.15 (56) 26.96 ± 1.00 (73) B C D ŁOCK 0137 40.15 ± 1.07 (50) 34.38 ± 2.11 (66) 26.88 ± 1.54 (73) B C D ŁOCK 0140 58.13 ± 0.86 (42) 47.09 ± 1.84 (53) 29.29 ± 1.50 (71) B C D ŁOCK 0142 44.29 ± 0.97 (56) 37.41 ± 0.76 (63) 25.53 ± 0.65 (74) Average concentration (μg/ml) (decrease (%)) 47.19 (53) 43.16 (57) 28.06 (72) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) ZEN Detoxification the highest detoxification activity towards FUM. The most resistant mycotoxin was DON. Lactobacillus sp. showed varied ability for zearalenone (ZEN) We found that the detoxification of AFB , DON, FUM and detoxification. After only 6 h of incubation, a statistically sig- ZEN mycotoxins by the tested Lactobacillus sp. and nificant reduction of ZEN concentration was observed, which S. cerevisiae strains were similar and did not show significant ranged between 28 and 59% (mean, 43%) relative to the initial differences. In contrast, the T-2 compound was more suscep- mycotoxin concentration of 100 μg/ml. In subsequent hours tible to removal from the mixture by yeast monocultures at a of incubation, a further reduction in the mycotoxin concentra- significance level of p < 0.05comparedtobacterial tion was noted. After 24 h of incubation, the concentration of monocultures. ZEN was 27.39–60.05 μg/ml (mean, 40.43 μg/ml). Therefore, The analysis allowed selection of four bacterial strains and a reduction of 40–73% (mean, 57%) in relation to the initial two yeast strains characterised by the best detoxification ca- concentration of the ZEN was observed in monocultures in- pabilities of all analysed mycotoxins. These include Lact. cubated with Lactobacillus sp. rhamnosus ŁOCK 1087, Lact. reuteri ŁOCK 1092, Lact. S. cerevisiae also demonstrated the ability to reduce the plantarum ŁOCK 0860, Lact. pentosus ŁOCK 1094, concentration of ZEN, and after 6 h of incubation, a significant S. cerevisiae ŁOCK 0119 and S. cerevisiae ŁOCK 1042. decrease of 24–42% (average 34%) to level of 57.64– 75.60 μg/ml (mean, 65.55 μg/ml) was observed. In subse- quent hours of incubation, the concentration of the mycotoxin further reduced, and after 24 h, ZEN concentrations ranged Discussion between 41.88 and 55.84 μg/ml (average reduction of 52% of the initial concentration) (Table 9). Mycotoxin detoxification methods can be classified by modes Based on these results, we concluded that mycotoxin of action applied before or after harvesting raw plant mate- (AFB ,DON, FUM,T-2,ZEN) detoxification properties of rials, which are used for human and animal nutrition. During Lactobacillus sp. and S. cerevisiae were strain-dependent. tillage process of plants, good agricultural practice (GAP) Both bacteria and yeast strains, subjected to analysis, showed should be maintained, including crop rotation, cultivation of Probiotics & Antimicro. Prot. Table 8 Reduction of T-2 concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B C D Lactobacillus brevis ŁOCK 1093 100 73.32 ± 3.18 (27) 60.74 ± 0.41 (39) 50.10 ± 1.32 (50) B C D casei ŁOCK 0911 54.11 ± 1.21 (46) 45.04 ± 1.59 (55) 36.83 ± 0.46 (63) B C D casei ŁOCK 0915 51.86 ± 3.06 (48) 43.24 ± 0.96 (57) 35.90 ± 0.48 (64) B C D paracasei ŁOCK 1091 60.17 ± 1.62 (40) 53.57 ± 1.83 (46) 48.24 ± 0.83 (52) B C D pentosus ŁOCK 1094 59.06 ± 2.45 (41) 43.47 ± 1.43 (57) 39.24 ± 1.41 (61) B C D plantarum ŁOCK 0860 48.62 ± 0.03 (52) 39.08 ± 0.17 (61) 31.46 ± 0.71 (69) B C D plantarum ŁOCK 0862 61.76 ± 3.04 (38) 48.28 ± 1.48 (52) 42.71 ± 1.07 (57) B C D reuteri ŁOCK 1092 53.86 ± 3.18 (46) 39.99 ± 1.20 (60) 31.09 ± 0.69 (69) B C D reuteri ŁOCK 1096 55.93 ± 3.57 (44) 42.39 ± 1.11 (58) 36.41 ± 0.72 (64) B C D rhamnosus ŁOCK 1087 48.25 ± 1.24 (52) 38.18 ± 1.37 (62) 26.76 ± 1.30 (73) B C D rhamnosus ŁOCK 1088 58.30 ± 4.13 (52) 49.88 ± 1.20 (51) 38.10 ± 1.42 (62) B C D rhamnosus ŁOCK 1089 63.08 ± 1.45 (37) 51.25 ± 1.99 (49) 45.78 ± 0.56 (54) Average concentration (μg/ml) (decrease (%)) 57.36 (43) 46.66 (54) 39.01 (61) A B C D S. cerevisiae ŁOCK 0068 100 53.66 ± 1.37 (46) 38.96 ± 2.43 (61) 31.83 ± 1.64 (68) B C D ŁOCK 0113 47.78 ± 1.34 (52) 40.04 ± 1.44 (60) 32.25 ± 0.77 (68) B C D ŁOCK 0119 46.92 ± 2.32 (53) 37.57 ± 0.53 (62) 31.06 ± 1.23 (69) B C D ŁOCK 0137 51.51 ± 1.34 (49) 37.73 ± 1.30 (62) 27.71 ± 1.40 (72) B C D ŁOCK 0140 54.98 ± 1.56 (45) 40.44 ± 1.75 (60) 32.62 ± 0.63 (67) B C D ŁOCK 0142 52.23 ± 0.92 (48) 37.36 ± 0.67 (62) 29.48 ± 0.86 (71) Average concentration (μg/ml) (decrease (%)) 51.18 (49) 38.68 (61) 30.54 (69) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) resistant plants, ploughing, irrigation, chemical and biological The AFB concentration in PBS solution was reduced by control of plant diseases and proper use of chemicals (e.g. the tested strains of bacteria to varying degrees, on average fungicides) [22, 34, 35]. When crops have already been har- 49.44% after the first 6 h of incubation, by another 4.82% vested, mycotoxin concentration can be reduced by adjust- during the next 6 h of incubation, and maintained within a ment of appropriate storage conditions (i.e. humidity and tem- range of 44.20–71.04 μg/ml (average reduction of 59.97%) perature), or by using detoxification treatments (physical, after 24 h of incubation. The greatest amount of AFB was chemical, biological) that can degrade, inactivate or decrease bound by bacteria strains after 6 h of incubation, suggesting the toxicity level of mycotoxins and ensure the nutritional that the adsorption of the mycotoxin by Lactobacillus is an value of food. Simultaneously, these detoxification methods immediate process. Haskard et al. (2001) tested the ability of should not introduce any major changes in production process eight Lactobacillus strains to bind AFB to bacterial surfaces technology [9, 34, 35]. using ELISA; their results were similar to ours [39]. Among other mycotoxin detoxification methods, microor- Liew et al. (2018), who also performed AFB binding as- ganisms, inter alia probiotic strains of Lactobacillus sp. and says with ELISA, confirmed results obtained by Haskard et al. S. cerevisiae, are used in case of mycotoxin contamination of (2001), and observed that live cells of Lactobacillus casei food and fodder [36]. Lactobacillus sp. are able to bind myco- Shirota were more efficient in binding mycotoxin, than heat- toxins mostly to cell wall peptidoglycans, polysaccharides and treated organisms [40]. Hernandez-Mendoza et al. (2009), teichoic acid, primarily through hydrophobic interactions, Huang et al. (2017) and Kumar et al. (2018) in their in vitro whereas S. cerevisiae are bind toxic metabolites of filamentous analysis using HPLC also showed the variable range of detox- fungi to the cell wall. In addition, microorganisms biodegrade ification level of AFB by Lactobacillus sp. In these studies, mycotoxins that prevent adsorption of these components inside AFB was bound by the tested strains of bacteria in the range the intestines on animals that feed on the food [24, 37, 38]. of 14–49%, 20.88–59.44% and 0–85%, respectively [41–43]. In this article, in vitro results demonstrated the ability to The variable binding ability of the Lactobacillus strains to reduce the concentration of mycotoxins: AFB , DON, FUM, AFB could be the result of differences in cell wall structure, 1 1 T-2, ZEN in PBS solution by 12 strains of bacteria from especially in terms of teichoic acid and peptidoglycan content Lactobacillus genus, and 6 S. cerevisiae strains. [44]. On the basis of the studies conducted, Gratz et al. (2005) Probiotics & Antimicro. Prot. Table 9 Reduction of ZEN concentration by Lactobacillus and S. cerevisiae strains Strain Time (h) 0 6 12 24 Concentration ± SD (μg/ml) (decrease (%)) A B C D Lactobacillus brevis ŁOCK 1093 100 63.61 ± 3.73 (36) 51.71 ± 2.57 (48) 43.17 ± 1.36 (57) B C D casei ŁOCK 0911 64.77 ± 3.78 (35) 57.41 ± 1.95 (43) 49.91 ± 2.33 (50) B C C casei ŁOCK 0915 63.69 ± 2.31 (36) 55.07 ± 1.82 (45) 51.22 ± 2.54 (49) B B C paracasei ŁOCK 1091 55.86 ± 2.35 (44) 50.67 ± 2.33 (49) 45.97 ± 1.38 (54) B B B pentosus ŁOCK 1094 41.21 ± 1.90 (59) 36.26 ± 3.88 (63) 32.22 ± 1.47 (67) B C D plantarum ŁOCK 0860 44.36 ± 1.29 (56) 35.42 ± 0.93 (65) 27.39 ± 1.46 (73) B C D plantarum ŁOCK 0862 69.93 ± 1.29 (30) 62.47 ± 1.73 (38) 56.63 ± 1.21 (43) B C D reuteri ŁOCK 1092 47.48 ± 1.04 (53) 38.26 ± 0.81 (62) 32.84 ± 1.25 (67) B B C reuteri ŁOCK 1096 72.45 ± 3.98 (28) 65.37 ± 2.31 (35) 60.06 ± 0.64 (40) B C D rhamnosus ŁOCK 1087 60.66 ± 1.78 (39) 52.93 ± 2.36 (47) 45.64 ± 1.54 (54) B C D rhamnosus ŁOCK 1088 58.00 ± 3.42 (42) 48.56 ± 0.65 (51) 41.25 ± 1.57 (58) B C D rhamnosus ŁOCK 1089 45.17 ± 1.70 (55) 35.05 ± 1.38 (65) 30.84 ± 0.82 (69) Average concentration (μg/ml) (decrease (%)) 57.27 (43) 49.10 (51) 43.10 (57) A B C D S. cerevisiae ŁOCK 0068 100 75.60 ± 2.88 (24) 55.78 ± 2.10 (44) 49.99 ± 1.25 (50) B C D ŁOCK 0113 68.47 ± 1.52 (32) 62.14 ± 1.80 (38) 55.84 ± 2.72 (44) B C D ŁOCK 0119 57.64 ± 1.67 (42) 47.16 ± 1.61 (53) 41.88 ± 1.21 (58) B C C ŁOCK 0137 58.95 ± 1.68 (41) 50.70 ± 2.14 (49) 47.47 ± 1.18 (53) B C D ŁOCK 0140 68.24 ± 2.23 (32) 55.59 ± 3.32 (44) 47.59 ± 1.31 (52) B C D ŁOCK 0142 64.38 ± 0.80 (36) 52.35 ± 2.83 (48) 46.03 ± 0.26 (54) Average concentration (μg/ml) (decrease (%)) 65.55 (34) 53.95 (46) 48.13 (52) *Values labelled by different capital letters were significantly different per analysed strain (p <0.05) showed that the binding of AFB by Lactobacillus sp. is a detoxification activity of 137 Lactobacillus strains, which rapid process, which was also confirmed in research conduct- wasalsoobservedinthestrainsusedinour study [49]. ed by Kumar et al.(2018), as well as in our studies [43, 45]. Similar results were obtained by Franco et al. (2011) and Moreover, tested S. cerevisiae strains varied in their reduction Zou et al. (2012), whereas in a study by García et al. (2018), of AFB concentration (on average 57.92% after 6 h of incu- even though they did not observe DON binding by bation, another 3.07% after 12 h of incubation, with a final Lactobacillus rhamnosus RC007, based on their results, they 3.69% reduction after 24 h), which is consistent with the re- concluded that Lact. rhamnosus contributed to counteract tox- sults obtained by Pizzolitto et al. (2012a) and Poloni et al. ic effect of DON and helped to maintain healthy gastrointes- (2017) [46, 47]. Strain-dependent AFB detoxification, by tinal tract of pigs [50–52]. In our studies, similarly low levels bacteria and yeast, was also observed in the results of studies of activity of S. cerevisiae were demonstrated, decreasing the by Pizzolitto et al. (2011) [48]. The detoxification activity of concentration of DON by 18.01%, on average, after 6 h of yeast strains to reduce the concentration of AFB was similar incubation, by 8.80% after another 6 h and an average of to that of the tested Lactobacillus, and what is also known in 33.03% of the initial concentration after 24 h of incubation. the case of yeast is that the cell wall components are also These results are in line with results obtained by Campagnollo responsible for the binding of the AFB mycotoxin [48]. et al. (2015) [53]. Based on these data, a significantly weaker On the basis of the results obtained, we conclude that DON binding activity by the analysed strains of microorgan- strains of Lactobacillus isolated from the intestinal content isms was also observed in comparison to other mycotoxins, of monogastric animals showed higher binding activity to- namely AF, ZEN or FUM, as also observed by Niderkorn wards DON, than did strains derived from plant silages. et al. (2007) and Campagnollo et al. (2015) [49, 53]. After 6 h of incubation, DON concentration was reduced by The 12 Lactobacillus strains analysed reduced FB and the bacterial strains tested by 5.76–21.64% (mean, 15.70%); FB mixture concentrations by the most significant amounts after 12 h, it was reduced by an average of another 7.39%; and among all mycotoxins tested. The FUM concentrations de- after 24 h, the concentration of DON was reduced by 19.28– creased on average by over 51.23% after 6 h of incubation, 39.37% (mean, 29.55%) of the initial concentration of the and a further 8.61% after 12 h. After 24 h of incubation, the mycotoxin. Niederkorn et al. (2007) showed low concentrations were maintained at an average level of Probiotics & Antimicro. Prot. 29.53 μg/ml (reduction by an average of 70.47% of the initial detoxification by S. cerevisiae strains in their studies and FUM concentration). The greatest reduction in concentration found the yeast’s reduction of the concentration of T-2 toxin occurred after 6 h, while the FUM binding process was ob- was also the result of mycotoxin binding to the yeast cell wall served up to 24 h of incubation, which is comparable to the [60]. results obtained by Abbès et al. (2016) [54]. Zhao et al. (2016) The ability of Lactobacillus strains to bind ZEN was sim- also showed that the process of FUM binding to the surface of ilar to that observed in the case of AFB and the detoxification Lactobacillus is rapid, as they observed the binding after 1 h level of the mycotoxin was mean of 56.90% after 24 h of of incubation [55]. In fact, Pizzolitto et al. (2012b) found that incubation. Our results showed that the greatest reduction in it was an immediate process, occurring after only a minute of ZEN concentrations, by 27.55–58.79% (mean, 42.73%), oc- incubation [56]. Deepthi et al. (2016) also reported high FB curred after 6 h of incubation. The ZEN concentrations were removal (61.7%) by Lact. plantarum MYS6, after 4 h of in- reduced further by an average of 8.17% after the next 6 h of cubation; furthermore, they observed that tested strain sup- incubation, finally reaching an average level of 43.10 μg/ml pressed fumonisin biosynthesis [57]. In our study, high after 24 h (an average 56.90% of the initial concentration of FUM detoxification levels exhibited by Lactobacillus sp., ZEN was bound by the bacterial strains). The capability of which was dependent on the strain tested and mycotoxin type, Lactobacillus strains to decrease ZEN concentration was also coincided with the results obtained by Niderkorn et al. (2007), found by Zhao et al. (2015); ZEN binding activities on the cell Abbès et al. (2016) and Zhao et al. (2016) [49, 54, 55]. walls of the 27 tested Lact. plantarum strains were weaker Moreover, Zhao et al. (2016) also noted that peptidoglycan compared to those of the strains we used in our study [61]. is primarily responsible for the binding of FUM, whereas Furthermore, Zhao et al. established that the binding of ZEN teichoic acid has little effect on mycotoxin binding [55]. On to the cell wall of Lactobacillus is an immediate process, the other hand, Martinez Tuppia et al. (2016), showed that FB which is consistent with the results we obtained [61]. detoxification might not only be a matter of adsorption, but Niderkorn et al. (2006), Čvek et al. (2012), Sangsila et al. also the result of biodegradation to other compounds (e.g. (2016) and Vega et al. (2017) showed comparable results from hydrolysed fumonisin B1, HFB ); however, this was not ob- their studies indicating great potential of Lactobacillus sp. to served for all analysed Lactobacillus sp. strains in their study reduce ZEN concentration [62–65]. We found that [58]. In our research, high and diverse activity of the analysed S. cerevisiae strains reduced ZEN concentration by 24.40– S. cerevisiae strains reducing FUM concentration was also 42.36% (average 34.45%) after 6 h of incubation, with an shown, and the results agree with those of Pizzolitto et al. additional 11.59% after 12 h and an average 5.82% after (2012b) [56]. Furthermore, Armando et al. (2013), who also 24 h. Similar results were obtained by Keller et al. (2015); observed high FB binding properties of S. cerevisiae yeast they concluded that at the beginning, large amounts of ZEN (strain RC016), observed that ability of binding the mycotoxin is bound to the yeast cell wall due to the higher concentration increases along with concentration [59]. Our results also of the mycotoxin in the environment [66]. The ZEN detoxifi- showed that the strains isolated from the feed were cation activity of the bacteria and yeast strains used in this characterised by a slightly lower ability to remove FUM study, was strain-dependent, a result also shown by (67.43% after 24 h of incubation) than achieved by baker’s Armando et al. (2012) and Zhao et al. (2015) [61, 67]. yeast (70.71–74.47% reduction of FUM concentration after 24 h of incubation) or strains isolated from a distillery envi- ronment (72.88% and 73.04% reduction of mycotoxin con- centration after 24 h of incubation). Conclusions The concentrations of T-2 toxin were decreased by the test- ed strains of Lactobacillus and S. cerevisiae to a high degree, Our data allowed us to observe the potential of Lactobacillus where reduction exceeded a mean of 60% after 24 h of incu- and S. cerevisiae to lower the concentrations of the myco- bation. After 6 h of incubation, the Lactobacillus strains re- toxins AFB , DON, FUM, T-2 and ZEN. We found that the duced the concentration of T-2 by 26.68–51.75% (average detoxification of these compounds is a rapid process. To a 42.64%), and after 24 h, it remained at a level of 31.09– great extent, the concentration of the toxins decreases within 50.10 μg/ml (average 39.01 μg/ml). Bacterial strains used in 6 h of incubation, and can last up to 24 h. Nevertheless, the our study showed higher T-2 binding affinity than shown by differences in concentrations were not so significant at a the four strains analysed by Zou et al. (2012) [51]. Only in prolonged period of 24-h incubation. relation to the T-2 toxin did the six S. cerevisiae strains show The Lactobacillus and S. cerevisiae strains analysed significantly stronger detoxification properties (average T-2 showed detoxification properties towards mycotoxins binding after 24 h of incubation was 69.18%) than the tested (AFB ,DON,FUM,T-2,ZEN)inthisinvitrostudy. bacterial strains (average decontamination rate after 24 h was Therefore, these strains, after further investigation, could be 61.45%). Zou et al. (2015) obtained a similar level of T-2 used as food and feed additives to detoxify contaminating Probiotics & Antimicro. Prot. 8. Ji C, Fan Y, Zhao L (2016) Review on biological degradation of mycotoxins, which pose potential threats to human and animal mycotoxins. Animal Nutr 2(3):127–133. https://doi.org/10.1016/j. health. aninu.2016.07.003 9. Zacharisova M, Dzuman Z, Veprikova Z, Hajkova K, Jiru M, Acknowledgements We would like to thank the National Centre for Vaclavikova M, Zacharisaova A, Pospichalova M, Florian M, Research and Development for the financial support of publication of this Hajslova J (2014) Occurrence of multiple mycotoxins in paper within the project PBS3/A8/32/2015 realised within the framework European feedingstuffs, assessment of dietary intake by farm ani- of the Program of Applied Studies. mals. Anim Feed Sci Technol 193:124–140. https://doi.org/10. 1016/j.anifeedsci.2014.02.007 10. Neme K, Mohammed A (2017) Mycotoxin occurrence in grains Funding This research was funded by the National Centre for Research and the role of postharvest management as a mitigation strategies. and Development within the project PBS3/A8/32/2015. A review. Food Control 78:412–425. https://doi.org/10.1016/j. foodcont.2017.03.012 Compliance with Ethical Standards 11. Kosicki R, Błajet-Kosicka A, Grajewski J, Twarużek M (2016) Multiannual mycotoxin survey in feed materials and feedingstuffs. Conflict of Interest The authors declare that they have no conflict of Anim Feed Sci Technol 215:165–180. https://doi.org/10.1016/j. interest. anifeedsci.2016.03.012 12. European Parliament and the Council (2002) Directive 2002/32/EC of the European Parliament and of the Council of 7 May 2002 on Ethical Approval This article does not contain any studies with human undesirable substances in animal feed - Council statement. https:// participants or animals performed by any of the authors. eur-lex.europa.eu/resource.html?uri=cellar:aca28b8c-bf9d-444f- Open Access This article is distributed under the terms of the Creative b470-268f71df28fb.0004.02/DOC_1&format=PDF. Accessed 06 Commons Attribution 4.0 International License (http:// April 2018 creativecommons.org/licenses/by/4.0/), which permits unrestricted use, 13. The Commission Of The European Communities (2006) distribution, and reproduction in any medium, provided you give appro- Commission Regulation (EC) No 1881/2006 of 19 December priate credit to the original author(s) and the source, provide a link to the 2006 setting maximum levels for certain contaminants in food- Creative Commons license, and indicate if changes were made. stuffs. http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri= CELEX:32006R1881&from=en. Accessed 06 April 2018 14. The Commission Of The European Communities (2006) Commission recommendation 2006/576/EC of 17 August 2006 Publisher’sNote Springer Nature remains neutral with regard to jurisdic- on the presence of deoxynivalenol, zearalenone, ochratoxin A, T- tional claims in published maps and institutional affiliations. 2 and HT-2 and fumonisins in products intended for animal feed- ing. http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri= CELEX:32006H0576&from=EN. Accessed 06 April 2018 15. Medina A, Magan N (2011) Temperature and water activity effects on production of T-2 and HT-2 by Fusarium langsethiae strains References from north European countries. Food Microbiol 28:392–398. ht tps://doi.org/10.1016/j.fm.2010.09.012 1. Smith MC, Madec S, Coton E, Hymery N (2016) Natural co- 16. Milani JM (2013) Ecological conditions affecting mycotoxin pro- occurrence of mycotoxins in foods and feeds and their in vitro duction in cereals: a review. Vet Med (Praha) 58(8):405–411. combined toxicological effects. Toxins 8(4):94. https://doi.org/10. https://doi.org/10.17221/6979-VETMED 3390/toxins8040094 17. The Commission Of The European Communities (2013) 2. Alshannaq A, Yu JH (2017) Occurrence, toxicity, and analysis of Commission Recommendation 2013/165/EU of 27 March 2013 major mycotoxins in food. Int J Environ Res Public Health 14(6): on the presence of t-2 and ht-2 toxin in cereals and cereal products. 632. https://doi.org/10.3390/ijerph14060632 http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri= 3. Liew WPP, Mohd-Redzwan S (2018) Mycotoxin: its impact on gut CELEX:32013H0165&from=EN. Accessed 06 April 2018 health and microbiota. Front Cell Infect Microbiol 8(60). https:// 18. Nugmanov A, Beishova I, Kokanov S, Lozowicka B, Kaczynski P, doi.org/10.3389/fcimb.2018.00060 Konecki R, Snarska K, Wołejko E, Sarsembayeva N, Abdigaliyeva 4. Gallo A, Giuberti G, Frisvad JC, Bertuzzi T, Nielsen KF (2015) T (2018) Systems to reduce mycotoxin contamination of cereals in Review on mycotoxin issues in ruminants: occurrence in forages, the agricultural region of Poland and Kazakhstan. Crop Prot 106: effects of mycotoxin ingestion on health status and animal perfor- 64–71. https://doi.org/10.1016/j.cropro.2017.12.014 mance and practical strategies to counteract their negative effects. 19. The European Parliament And The Council Of The European Toxins 7(8):3057–3111. https://doi.org/10.3390/toxins7083057 Union (2017) Regulation (EU) 2017/625 of the European 5. Pinotti L, Ottoboni M, Giromini C, Dell’Orto V, Cheli F (2016) Parliament and of the Council of 15 March 2017 on official controls Mycotoxin contamination in the EU feed supply chain: a focus on and other official activities performed to ensure the application of cereal byproducts. Toxins 8(2):45. https://doi.org/10.3390/ food and feed law, rules on animal health and welfare, plant health toxins8020045 and plant protection products. https://eur-lex.europa.eu/legal- 6. Freire L, Sant’Ana AS (2018) Modified mycotoxins: an updated content/EN/TXT/PDF/?uri=CELEX:32017R0625&from=EN. review on their formation, detection, occurrence, and toxic effects. Accessed 06 April 2018 Food Chem Toxicol 111:189–205. https://doi.org/10.1016/j.fct. 20. Aiko V, Mehta L (2015) Occurrence, detection and detoxification of 2017.11.021 mycotoxins. J Biosci 45(5):943–954. https://doi.org/10.1007/ s12038-015-9569-6 7. Streit E, Schatzmayr G, Tassis P, Tzika E, Marin D, Taranu I, Tabacu C, Nicolau A, Aprodu I, Puel O, Oswald IP (2012) 21. Loi M, Fanelli F, Liuzzi VC, Logrieco AF, Mulè G (2017) Current situation of mycotoxin contamination and co-occurrence Mycotoxin biotransformation by native and commercial enzymes: present and future perspectives. Toxins 9(4):111. https://doi.org/10. in animal feed-focus on Europe. Toxins 4(10):788–809. https:// doi.org/10.3390/toxins4100788 3390/toxins9040111 Probiotics & Antimicro. Prot. 22. Zhu Y, Hassan YI, Lepp D, Shao S, Zhou T (2017) Strategies and 40. Liew WPP, Nurul-Adilah Z, Than LTL, Mohd-Redzwan S (2018) methodologies for developing microbial detoxification systems to The binding efficiency and interaction of Lactobacillus casei mitigate mycotoxins. Toxins 9(4):130. https://doi.org/10.3390/ Shirota toward aflatoxin B . Front Microbiol 9(1503). https://doi. toxins9040130 org/10.3389/fmicb.2018.01503 41. Hernandez-Mendoza A, Garcia HS, Steele JL (2009) Screening of 23. Juodeikiene G, Bartkiene E, Cernauskas D, Cizeikiene D, Zadeike Lactobacillus casei strains for their ability to bind aflatoxin B . D, Lele V, Bartkevics V (2018) Antifungal activity of lactic acid Food Chem Toxicol 47:1064–1068. https://doi.org/10.1016/j.fct. bacteria and their application for Fusarium mycotoxin reduction in 2009.01.042 malting wheat grains. LWT-Food Sci Technol 89:307–314. https:// doi.org/10.1016/j.lwt.2017.10.061 42. Huang L, Duan C, Zhao Y, Gao L, Niu C, Xu J, Li S (2017) Reduction of aflatoxin b toxicity by lactobacillus plantarum c88: 24. El-Nezami H, Kankaanpaa P, Salminen S, Ahokas J (1998) Ability 1 a potential probiotic strain isolated from Chinese traditional of dairy strains of lactic acid bacteria to bind a common food car- fermented food Btofu^. PLoS One 12(1):e0170109. https://doi. cinogen, aflatoxin B1. Food Chem Toxicol 36:321–326 org/10.1371/journal.pone.0170109 25. FAO/WHO (2002) Guidelines for the evaluation of probiotics in 43. Kumar SS, Bashisht A, Venkateswaran G, Hariprasad P, Gayathri D food. Food and Agriculture Organization of the United Nations (2018) Characterization of novel Lactobacillus fermentum from and World Health Organization Working Group Report. http:// curd samples of indigenous cows from Malnad region, Karnataka, www.fao.org/3/a-a0512e.pdf. Accessed 24 March 2018 for their aflatoxin B binding and probiotic properties. Probiotics 26. Vinderola G, Ritieni A (2015) Role of probiotics against myco- Antimicrob Proteins. https://doi.org/10.1007/s12602-018-9479-7 toxins and their deleterious effects. J Food Res 4(1):10–21. 44. Hernandez-Mendoza A, Guzman-de-Peña D, Garcia HS (2009) https://doi.org/10.5539/jfr.v4n1p10 Key role of 383 teichoic acids on aflatoxin B 384 binding by 27. Śliżewska K, Chlebicz A (2017) Strains of lactic acid bacteria from probiotic bacteria. J Appl Microbiol 107:395–403. https://doi.org/ the genus Lactobacillus. Patent Application no 422603 10.1111/j.1365-3852672.2009.04217.x 28. Aflatoxin B from Aspergillus flavus, Sigma-Aldrich. https://www. 45. Gratz S, Mykkänen H, El-Nezami H (2005) Aflatoxin B binding sigmaaldrich.com/catalog/product/sigma/a6636?lang=pl&region= by a mixture of Lactobacillus and Propionibacterium: in vitro ver- PL. Accessed 21 March 2018 sus ex vivo. J Food Prot 68(11):2470–2474. https://doi.org/10.4315/ 29. Deoxynivalenol, Sigma-Aldrich. https://www.sigmaaldrich.com/ 0362-028X-68.11.2470 catalog/product/sial/32943?lang=pl&region=PL. Accessed 21 46. Pizzolitto RP, Armando MR, Combina M, Cavaglieri LR, Dalcero March 2018 AM, Salvano MA (2012) Evaluation of Saccharomyces cerevisiae 30. Fumonisin B , Sigma-Aldrich. https://www.sigmaaldrich.com/ strains as probiotic agent with aflatoxin B adsorption ability for use catalog/product/sial/34139?lang=pl&region=PL&cm_sp=Insite-_- in poultry feedstuffs. J Environ Sci Health B 47(10):933–941. prodRecCold_xorders-_-prodRecCold2-1. Accessed 21 https://doi.org/10.1080/03601234.2012.706558 March 2018 47. Poloni V, Salvato L, Pereyra L, Oliveira A, Rosa C, Cavaglieri L, 31. Fumonisin B , Sigma-Aldrich. https://www.sigmaaldrich.com/ Keller KM (2017) Bakery by-products based feeds borne- catalog/product/sial/34142?lang=pl&region=PL&cm_sp=Insite-_- Saccharomyces cerevisiae strains with probiotic and antimycotoxin prodRecCold_xorders-_-prodRecCold2-2. Accessed 21 effects plus antibiotic resistance properties for use in animal pro- March 2018 duction. Food Chem Toxicol 107:630–636. https://doi.org/10.1016/ 32. T-2 toxin, Sigma-Aldrich. https://www.sigmaaldrich.com/catalog/ j.fct.2017.02.040 product/sigma/t4887?lang=pl&region=PL. Accessed 21 48. Pizzolitto RP, Bueno DJ, Armando MR, Cavaglieri L, Dalcero AM, March 2018 Salvano MA (2011) Binding of aflatoxin B to lactic acid bacteria 33. Zearalenone, Sigma-Aldrich. https://www.sigmaaldrich.com/ and Saccharomyces cerevisiae in vitro: a useful model to determine catalog/product/sigma/z2125?lang=pl&region=PL. Accessed 21 the most efficient microorganism. In: Guevara-Gonzalez RG (ed) March 2018 Aflatoxins. Biochemistry and molecular biology, InTech. https:// 34. Devreese M, De Backer P, Croubels S (2013) Different methods to doi.org/10.5772/23717. https://www.intechopen.com/books/ counteract mycotoxin production and its impact on animal health. aflatoxins-biochemistry-and-molecular-biology/binding-of- Vlaams Diergeneeskd Tijdschr 82(4):181–190 aflatoxin-b1-to-lactic-acid-bacteria-and-saccharomyces-cerevisiae- 35. Karlovsky P, Suman M, Berthiller F, De Meester J, Eisenbrand G, in-vitro-a-useful-model. Accessed 28 March 2018 Perrin I, Oswald IPm Speijers G, Chiodini A, Recker T, Dussort P 49. Niderkorn V, Morgavi DP, Pujos E, Tissandier A, Boudra H (2007) (2016) Impact of food processing and detoxification treatments on Screening of fermentative bacteria for their ability to bind and mycotoxin contamination. Mycotoxin Res 32(4):179–205. https:// biotransform deoxynivalenol, zearalenone and fumonisins in an doi.org/10.1007/s12550-016-0257-7 in vitro simulated corn silage model. Food Addit Contam 24(4): 36. Moslehi-Jenabian S, Pedersen LL, Jespesen L (2010) Beneficial 406–415. https://doi.org/10.1080/02652030601101110 effects of probiotic and food borne yeasts on human health. 50. Franco TS, Garcia S, Hirooka EY, Ono YS, Santos JS (2011) Lactic Nutrients 2(4):449–473. https://doi.org/10.3390/nu2040449 acid bacteria in the inhibition of Fusarium graminearum and 37. Śliżewska K, Smulikowska S (2011) Detoxification of aflatoxin B deoxynivalenol detoxification. J Appl Microbiol 111:739–748. and change in microflora pattern by probiotic in vitro fermentation https://doi.org/10.1111/j.1365-2672.2011.05074.x of broiler feed. Anim Feed Sci Technol 20:300–309. https://doi.org/ 51. Zou ZY, He ZF, Li HJ, Han PF, Meng X, Zhang Y, Zhou F, Ouyang 10.22358/jafs/66187/2011 KP, Chen XY, Tang J (2012) In vitro removal of deoxynivalenol and 38. Vila-Donat P, Marin S, Sachis V, Ramos AJ (2018) A review of the T-2 toxin by lactic acid bacteria. Food Sci Biotechnol 21(6):1677– mycotoxin adsorbing agents, with an emphasis on their multi- 1683. https://doi.org/10.1007/s10068-012-0223-x binding capacity, for animal feed decontamination. Anim Feed 52. García GR, Payros D, Pinton P, Dogi CA, Laffitte J, Neves M, Sci Technol 114:246–259. https://doi.org/10.1016/j.fct.2018.02. González Pereyra ML, Cavaglieri LR, Oswald IP (2018) 044 Intestinal toxicity of deoxynivalenol is limited by Lactobacillus rhamnosus RC007 in pig jejunum explants. Arch Toxicol 92: 39. Haskard CA, El-Nezmi HS, Kankaanpää PE, Salminen S, Ahokas 983–993. https://doi.org/10.1007/s00204-017-2083-x JT (2001) Surface binding of aflatoxin B by lactic acid bacteria. 53. Campagnollo FB, Franco LT, Rottinghaus GE, Kobashigawa E, Appl Environ Microbiol 67(7):3086–3091. https://doi.org/10.1128/ Ledoux DR, Daković A, Oliveira CAF (2015) In vitro evaluation AEM.67.7.3086-3091.2001 Probiotics & Antimicro. Prot. of the ability of beer fermentation residue containing 60. Zou Z, Sun J, Huang F, Feng Z, Li M, Shi R, Ding J, Li H (2015) Saccharomyces cerevisiae to bind mycotoxins. Food Res Int 77: In vitro removal of T-2 toxin by yeasts. J Food Safety 35:544–550. 643–648. https://doi.org/10.1016/j.foodres.2015.08.032 https://doi.org/10.1111/jfs.12204 61. Zhao L, Jin H, Zhang R, Ren H, Zhang X, Yu G (2015) 54. Abbès S, Salah-Abbès JB, Jebali R, Younes RB, Oueslati R (2016) Detoxification of zearalenone by three strains of Lactobacillus Interaction of aflatoxin B and fumonisin B in mice causes 1 1 plantarum from fermented food in vitro. Food Control 54:158– immunotoxicity and oxidative stress: possible protective role using 164. https://doi.org/10.1016/j.foodcont.2015.02.003 lactic acid bacteria. J Immunotoxicol 13(1):46–54. https://doi.org/ 62. Niderkorn V, Boudra H, Morgavi DP (2006) Binding of Fusarium 10.3109/1547691X.2014.997905 mycotoxins by fermentative bacteria in vitro. J Appl Microbiol 101: 55. Zhao H, Wang X, Zhang J, Zhang J, Zhang B (2016) The mecha- 849–856. https://doi.org/10.1111/j.1365-2672.2006.02958.x nism of Lactobacillus strains for their ability to remove fumonisins 63. Čvek D, Markov K, Frece J, Friganović M, Duraković L, Delaš F B and B . Food Chem Toxicol 97:40–46. https://doi.org/10.1016/j. 1 2 (2012) Adhesion of zearalenone to the surface of lactic acid bacteria fct.2016.08.028 cells. Croat J Food Tech Biotech Nutr 7:49–52 56. Pizzolitto RP, Salvano MA, Dalcero AM (2012) Analysis of 64. Sangsila A, Faucet-Marquis V, Pfohl-Leszkowicz A, Itsaranuwat P fumonisin B removal by microorganisms in co-occurrence with (2016) Detoxification of zearalenone by Lactobacillus pentosus aflatoxin B and the nature of the binding process. Int J Food strains. Food Control 62:187–192. https://doi.org/10.1016/j. Microbiol 156:214–221. https://doi.org/10.1016/j.ijfoodmicro. foodcont.2015.10.031 2012.03.024 65. Vega MF, Dieguez SN, Riccio B, Aranguren S, Giordano A, 57. Deepthi BV, Poornachandra Rao K, Chennapa G, Naik MK, Denzoin L, Soraci AL, Tapia MO, Ross R, Apás A, González SN Chandrashekara KT, Sreenivasa MY (2016) Antifungal attributes (2017) Zearalenone adsorption capacity of lactic acid bacteria iso- of Lactobacillus plantarum MYS6 against fumonisin producing lated from pigs. Braz J Microbiol 48(4):715–723. https://doi.org/ Fusarium proliferatum associated with poultry feeds. PLoS One 10.1016/j.bjm.2017.05.001 11(6):e0155122. https://doi.org/10.1371/journal.pone.0155122 66. Keller L, Abrunhosa L, Keller K, Rosa CA, Cavaglieri L, Venâncio 58. Martinez Tuppia C, Atanasova-Penichon V, Chéreau S, Ferrer N, A (2015) Zearalenone and its derivatives α-zearalenol and β- Marchegay G, Savoie J, Richard-Forget F (2017) Yeast and bacteria zearalenol decontamination by Saccharomyces cerevisiae strains from ensiled high moisture maize grains as potential mitigation isolated from bovine forage. Toxins 7(8):3297–3308. https://doi. agents of fumonisin B . J Sci Food Agric 97:2443–2452. https:// org/10.3390/toxins7083297 doi.org/10.1002/jsfa.8058 67. Armando MR, Pizzolitto RP, Dogi CA, Cristofolini A, Merkis C, 59. Armando M, Galvagno M, Dogi C, Cerrutti P, Dalcero A, Poloni V, Dalcero AM, Cavaglieri LR (2012) Adsorption of ochra- Cavaglieri L (2013) Statistical optimization of culture conditions toxin A and zearalenone by potential probiotic Saccharomyces for biomass production of probiotic gut-borne Saccharomyces cerevisiae strains and its relations with cell wall thickness. J Appl cerevisiae strain able to reduce fumonisin B . J Appl Microbiol Microbiol 113:256–264. https://doi.org/10.1111/j.1365-2672.2012. 114:1338–1346. https://doi.org/10.1111/jam.12144 05331.x

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