Sting that DacA and DacB break the was bond of OTA. These h of incubation. amide higher than 30 immediately after 72results demonstrate that DacA and DacB possess OTA The kinetic parameters, Km and Vmax, have been 2.74 /mL and 73.53 ng/h/mg for degrading activity, plus the strain ANSB168 can eliminate OTA employing biodegradation. DacA and 1.14 /mL, and 42.74 ng/h/mg for DacB when determined at 37 C and optimal pH. The OTA degradation ratio elevated more than the incubation time. DacA and DacB have been capable to degrade 45 and 42 of OTA after 72 h, respectively (AZD1656 web Figure 4C). two.4. Degraded Solution Identification of DacA and DacBHigh-performance liquid chromatography (HPLC) analysis indicated that the DacA and DacB degradation merchandise have been eluted as a peak with a retention time of 6.7 min that had the same transition time of OT (Figure five), suggesting that DacA and DacB break the amide bond of OTA. These final results demonstrate that DacA and DacB possess OTAdegrading activity, along with the strain ANSB168 can remove OTA employing biodegradation.Int. J. Mol. Sci. 2021, 22, 12059 Int. J. Mol. Sci. 2021, 22, x FOR PEER REVIEW66 of 19 ofFigure five. Determination of OTA and OT HPLC: (A) OTA normal; (B) OT regular; (C) OTA Figure 5. Determination of OTA and OT by by HPLC: (A) OTA standard; (B) OT normal; (C) OTA regular processed by DacA; and (D) OTA normal processed by DacB. common processed by DacA; and (D) OTA common processed by DacB.two.5. Ameliorative Effects of Bacillus subtilis ANSB168 in Laying Hens two.5. Ameliorative Effects of Bacillus subtilis ANSB168 in Laying Hens two.five.1. Production Efficiency two.five.1. Production Efficiency Feeding layers with OTA in the concentration of 250 /kg had negative effects on Feeding layers with OTA in the concentration of 250 g/kg had damaging effects on laying hens’ Thiacetazone In Vivo measured overall performance parameters (i.e., egg production ratio, average egg laying hens’ measured efficiency parameters (i.e., egg production ratio, average egg weight, everyday egg production, feed/egg ratio, and typical every day feed intake). In Figure 6A, weight, day-to-day egg production, feed/egg ratio, and average everyday feed intake). In FigureInt. J. Mol. Sci. 2021, 22, x FOR PEER Critique Int. J. Mol. Sci. 2021, 22,7 of 19 7 of6A, the day-to-day feed intake with the OTAfed group as well as the OTAANSB168 group was sig nificantly decreased compared together with the handle group (p 0.01). Both the egg production the day-to-day feed intake from the OTA-fed group and also the OTAANSB168 group was significantly decreased 6B, p = 0.059) the manage group (p 0.01). Each the egg = 0.099) of ratio ratio (Figure compared withand the each day egg production (Figure 6C, p production the (Figure 6B, p = 0.059) downward trend compared (Figure 6C, p = 0.099) of compared OTAfed group had a and also the every day egg production using the manage. When the OTA-fed group had a downward trend compared with all the control. When compared using the handle with the manage group, the egg production ratio as well as the every day egg production decreased group, the egg production ratio plus the each day egg production decreased five.49 and six.25 , 5.49 and six.25 , respectively. Despite the fact that not statistically important because of the variable respectively. Despite the fact that not statistically considerable as a result of variable variations within variations within groups, the OTAfed group’s feed/egg ratio improved by 5.28 (Fig groups, the OTA-fed group’s feed/egg ratio increased by five.28 (Figure 6D) and typical ure 6D) and average egg weight decreased 0.71 (Figure 6E) compared together with the co.