Zearalenone (ZEN) is a mycotoxin produced by Fusarium species, which is one of the main animal feed contaminants causing reproductive disorders in livestock. The aim of this study was to evaluate the probiotic characteristics and ZEN removal ability of a Bacillus licheniformis strain CK1. The probiotic properties, including acidic tolerance, bile salt tolerance, adherence capability, and anti-pathogenic activities of CK1 were evaluated. CK1 survived after incubation at pH 2.0 or 3.0 for 3 h, grew well in LB broth containing 0.3% oxgall, possessed adherence capability to Caco-2 cells, and inhibited the growth of Escherichia coli O157:H7 and Listeria monocytogenes. The ZEN removal ability of CK1 was compared with a mineral mycotoxin-adsorbing agent, hydrated sodium calcium aluminosilicate (HSCAS), and a well-characterized biological mycotoxin-adsorbing agent, Lactobacillus rhamnosus GG (LGG). At 37°C in phosphate-buffered saline (PBS, pH 7.0) containing 5 μg mL-1 of ZEN, the ZEN removal percentage of CK1 was 73.0%, which was significantly higher than that of HSCAS and LGG (45.9% and 48.4%, respectively). In the pH range of 2.5–8.0, CK1 removed up to 65% of ZEN. At temperatures between 4 and 42°C, CK1 removed more than 75% of ZEN. In the adsorption stability analysis, the amounts of ZEN removed by CK1 was over 30% even after five consecutive rounds of washing procedures. These findings demonstrated that CK1 displayed probiotic characteristics and removed ZEN effectively. Therefore, CK1 has a great potential for the development of feed additive to remove ZEN.

BackgroundMycotoxins are secondary metabolites produced by filamentous fungi that can contaminate agricultural crops in the field as well as during harvest, transportation, processing, or storage. Zearalenone (ZEN), a non-steroidal estrogenic mycotoxin, produced by Fusarium species, has been shown to be associated with reproductive disorders in farm animals and to a lesser extent in hyperoestrogenic syndromes in humans. Thus, the decontamination of ZEN in foods and feeds is an important issue.ResultsIn this study, the gene encoding ZHD101, a ZEN-degrading enzyme produced by Clonostachys rosea IFO 7063, was cloned into an Escherichia coli–Lactobacillus shuttle vector, pNZ3004, and the resultant plasmid pNZ-zhd101 was then introduced via electroporation into Lactobacillus reuteri Pg4, a probiotic strain isolated from the gastrointestinal tract of broilers. The transformed strain L. reuteri pNZ-zhd101 acquired the capacity to degrade ZEN. In addition, the production of recombinant ZHD101 did not affect cell growth, acid and bile salt tolerance, and had only a minor effect on the adhesion ability of L. reuteri pNZ-zhd101.ConclusionsTo the best of our knowledge, this is the first report of successful expression of a ZEN-degrading enzyme by intestinal lactobacilli.

Fruits have been widely considered as the default "health foods" because they contain numerous vitamins and minerals needed to sustain human health. Fermentation strategies have been utilized to enhance the nutritive and flavor features of healthy and readily consumable fruit products while extending their shelf lives. A traditional fermented multi-fruit beverage was made from five fruits including kiwi, guava, papaya, pineapple, and grape fermented by Saccharomyces cerevisiae along with lactic acid bacteria and acetic acid bacteria. The immunomodulatory properties of the fermented multi-fruit beverage, in vivo nonspecific and ovalbumin (OVA)-specific immune response experiments using female BALB/c mice were performed. Administration of the fermented multi-fruit beverage reduced the calorie intake, thus resulting in a less weight gain in mice compared to the water (placebo)-fed mice. In the nonspecific immune study model, the fermented multi-fruit beverage enhanced phagocytosis and T cell proliferation but did not affect B cell proliferation and immunoglobulin G (IgG) production. Analysis of cytokine secretion profile also revealed that the fermented multi-fruit beverage enhanced proinflammatory cytokines interleukin (IL)-6, tumor necrosis factor (TNF)-α, and T helper (Th)1-related cytokine interferon (IFN)-γ production, thus creating an immunostimulatory effect. Nonetheless, in the specific immune study model, the results showed that the fermented multi-fruit beverage decreased the production of proinflammatory cytokines IL-6 and TNF-α production in OVA-immunized mice. Moreover, it also caused a decrease in the production of anti-OVA IgG1, which was accompanied by a decrease in Th2-related cytokines IL-4 and IL-5 production and an increase in Th1-related cytokine IFN-γ production, indicating that it may have the potential to shift the immune system from the allergen-specific Th2 responses toward Th1-type responses. The results indicate that fermented multi-fruit beverage has the potential to modulate immune responses both in a nonspecific and specific manners.

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