Diet xylo-oligosaccharide supplementation improves growth performance, immune function, and intestinal health of broilers.

The effects of xylo-oligosaccharides (XOS) on broiler growth performance, immune function, and intestinal health were investigated. A total of 540 one-d-old Arbor Acres Plus broilers were randomly divided into 5 groups with 6 replicates per group and 18 chickens per replicate. Broilers in the control (CON) group received a corn-soybean meal based basal diet, those in the antibiotics (ANT) group received the basal diet plus 500 mg/kg oxytetracycline, and those in XOS groups received the basal diet plus 150, 300, or 450 mg/kg XOS. Compared with CON, the body weight at 42 d and average daily gain from 1 to 42 d were significantly increased in the 150, 450 mg/kg XOS-added and ANT groups (P = 0.018), and the relative expression of claudin-1 and ZO-1 mRNA in the ileum was significantly higher in the 300 and 450 mg/kg XOS-added groups (P < 0.001). The feed conversion ratios (P < 0.001) and abdominal fat rates (P = 0.012) of broilers from 1 to 42 d of age were significantly lower in all XOS-added groups than in the control group. Splenic index (P = 0.036) and bursa of Fabricius index (P = 0.009) were significantly better in the ANT group and each XOS-added group than in the control group. Compared to CON and ANT, serum IgA (P = 0.007) and IgG (P = 0.002) levels were significantly higher in the 300 mg/kg XOS-added group, and the relative abundance of short-chain fatty acid-producing genera (Alistipes) was also significantly higher (P < 0.001). Meanwhile, ileal villus height (P < 0.001) and ratio of villus height to crypt depth (V:C) (P = 0.001) were significantly increased in XOS-added broilers. In analysis of relationships between cecal microbes and the physical barrier of the gut, [Ruminococcus]_torques_group was positively correlated with mRNA expression of ileal ZO-1 and claudin-1 (P < 0.05), and Bacteroides was positively correlated with increased ileal villus height and V:C (P < 0.05). Overall, XOS addition to broiler diets improved growth performance, promoted intestinal health by enhancing intestinal barrier function and regulating cecal microbiota diversity, and had positive effects on immunity.

Pleurotus eryngii polysaccharides alleviate aflatoxin B1-induced liver inflammation in ducks involving in remodeling gut microbiota and regulating SCFAs transport via the gut-liver axis.

Aflatoxin B1 (AFB1) is one of the most widespread contaminants in agricultural commodities. Pleurotus eryngii (PE) is widely used as a feed additive for its anti-inflammatory properties, and its major active substance is believed to be polysaccharides. This study aims to explore the underlying mechanism of dietary PE polysaccharides alleviating AFB1-induced toxicity in ducks. The major monosaccharide components of PE polysaccharides were identified as glucose, mannose, galactose, glucuronic acid, and fucose. The results showed that dietary PE polysaccharides could alleviate liver inflammation, alleviate intestinal barrier dysfunction, and change the imbalanced gut microbiota induced by AFB1 in ducks. However, PE polysaccharides failed to exert protective roles on the liver and intestine injury induced by AFB1 in antibiotic-treated ducks. The PE + AFB1-originated microbiota showed a positive effect on intestinal barrier and inflammation, the SCFAs transport via the gut-liver axis, and liver inflammation compared with the AFB1-originated microbiota in ducks. These findings provided a possible mechanism that PE polysaccharides alleviated AFB1-induced liver inflammation in ducks by remodeling gut microbiota, regulating microbiota-derived SCFAs transport via the gut-liver axis, and inhibiting inflammatory gene expressions in the liver, which may provide new insight for therapeutic methods against AFB1 exposure in animals.

Biodegradation of ochratoxin A by Brevundimonas diminuta HAU429: Characterized performance, toxicity evaluation and functional enzymes.

Ochratoxin A (OTA) is a notorious mycotoxin commonly contaminating food products worldwide. In this study, an OTA-degrading strain Brevundimonas diminuta HAU429 was isolated by using hippuryl-L-phenylalanine as the sole carbon source. The biodegradation of OTA by strain HAU429 was a synergistic effect of intracellular and extracellular enzymes, which transformed OTA into ochratoxin α (OTα) through peptide bond cleavage. Cytotoxicity tests and cell metabolomics confirmed that the transformation of OTA into OTα resulted in the detoxification of its hepatotoxicity since OTA but not OTα disturbed redox homeostasis and induced oxidative damage to hepatocytes. Genome mining identified nine OTA hydrolase candidates in strain HAU429. They were heterologously expressed in Escherichia coli, and three novel amidohydrolase BT6, BT7 and BT9 were found to display OTA-hydrolyzing activity. BT6, BT7 and BT9 showed less than 45 % sequence identity with previously identified OTA-degrading amidohydrolases. BT6 and BT7 shared 60.9 % amino acid sequence identity, and exhibited much higher activity towards OTA than BT9. BT6 and BT7 could completely degrade 1 µg mL-1 of OTA within 1 h and 50 min, while BT9 hydrolyzed 100 % of OTA in the reaction mixture by 12 h. BT6 was the most thermostable retaining 38 % of activity after incubation at 70 °C for 10 min, while BT7 displayed the highest tolerance to ethanal remaining 76 % of activity in the presence of 6 % ethanol. This study could provide new insights towards microbial OTA degradation and promote the development of enzyme-catalyzed OTA detoxification during food processing.

A NADPH-Dependent Aldo/Keto Reductase Is Responsible for Detoxifying 3-Keto-Deoxynivalenol to 3-epi-Deoxynivalenol in Pelagibacterium halotolerans ANSP101.

Deoxynivalenol (DON), primarily generated by Fusarium species, often exists in agricultural products. It can be transformed to 3-epi-deoxynivalenol (3-epi-DON), with a relatively low toxicity, via two steps. DDH in Pelagibacterium halotolerans ANSP101 was proved to convert DON to 3-keto-deoxynivalenol (3-keto-DON). In the present research, AKR4, a NADPH-dependent aldo/keto reductase from P. halotolerans ANSP101, was identified to be capable of converting 3-keto-DON into 3-epi-DON. Our results demonstrated that AKR4 is clearly a NADPH-dependent enzyme, for its utilization of NADPH is higher than that of NADH. AKR4 functions at a range of pH 5-10 and temperatures of 20-60 °C. AKR4 is able to degrade 89% of 3-keto-DON in 90 min at pH 7 and 50 °C with NADPH as the cofactor. The discovery of AKR4, serving as an enzyme involved in the final step in DON degradation, might provide an option for the final detoxification of DON in food and feed.

Curcumin alleviates zearalenone-induced liver injury in mice by scavenging reactive oxygen species and inhibiting mitochondrial apoptosis pathway.

Curcumin (CUR) is a compound extracted from turmeric that has a variety of functions including antioxidant and anti-inflammatory. As an estrogen-like mycotoxin, zearalenone (ZEN) not only attacks the reproductive system, but also has toxic effects on the liver. However, whether CUR can alleviate ZEN-induced liver injury remains unclear. This paper aims to investigate the protective effect of CUR against ZEN-induced liver injury in mice and explore the molecular mechanism involved. BALB/c mice were randomly divided into control (CON) group, CUR group (200 mg/kg b. w. CUR), ZEN group (40 mg/kg b. w. ZEN) and CUR+ZEN group (200 mg/kg b. w. CUR+40 mg/kg b. w. ZEN). 28 d after ZEN exposure and CUR treatment, blood and liver samples were collected for subsequent testing. The results showed that CUR reversed ZEN-induced hepatocyte swelling and necrosis in mice. It significantly reduced the serum alkaline phosphatase (ALP), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in mice (p < 0.05). In addition, CUR significantly reduced hepatic ROS, malondialdehyde, hydrogen peroxide and apoptosis levels in mice (p < 0.05). Quantitative RT-PCR and Western blot results showed that CUR significantly reduced the expression of Bax and Caspase3, and reversed the increase of Nrf2, HO-1 and NQO1 expression in the liver of mice induced by ZEN (p < 0.05). In conclusion, CUR alleviated ZEN-induced liver injury in mice by scavenging ROS and inhibiting the mitochondrial apoptotic pathway.

Alleviative Effect of Rutin on Zearalenone-Induced Reproductive Toxicity in Male Mice by Preventing Spermatogenic Cell Apoptosis and Modulating Gene Expression in the Hypothalamic-Pituitary-Gonadal Axis.

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin found in many agricultural products and can cause reproductive disorders, mainly affecting spermatogenesis in male animals. Rutin (RUT) is a natural flavonoid compound recognized for its significant antioxidant, anti-inflammatory and estrogenic properties. The present study aimed to determine the protective role of RUT against ZEN-induced reproductive toxicity in male mice. Twenty-four adult Kunming male mice were divided into four groups: control, RUT (500 mg/kg RUT), ZEN (10 mg/kg ZEN), ZEN + RUT (500 mg/kg RUT + 10 mg/kg ZEN), with six replicates per treatment. The results indicated that RUT mitigated ZEN-induced disruption in spermatogenic cell arrangement, decreased spermatozoa count, and increased sperm mortality in the testes. RUT significantly restored ZEN-induced reduction in T, FSH, LH, and E2 serum levels. Moreover, RUT mitigated ZEN-induced apoptosis by increasing the mRNA expression level of bcl-2, decreasing the mRNA expression level of kiss1-r, and decreasing the protein expression level of caspase 8 in reproductive tissues. These findings indicate the protective role of RUT against ZEN-induced reproductive toxicity in male mice by regulating gonadotropin and testosterone secretions to maintain normal spermatogenesis via the HPG axis, which may provide a new application direction for RUT as a therapeutic agent to mitigate ZEN-induced reproductive toxicity.

Effective Degradation of Free Gossypol in Defatted Cottonseed Meal by Bacterial Laccases: Performance and Toxicity Analysis.

Cottonseed meal (CSM) is the major by-product of the cottonseed oil extraction process with high protein content. However, the presence of free gossypol (FG) in CSM severely restricts its utilization in the food and animal feed industries. The development of a biological strategy for the effective removal of FG in CSM has become an urgent need. In this study, three bacterial laccases including CotA from Bacillus licheniformis, CueO from Escherichia coli, and LcLac from Loigolactobacillus coryniformis were heterologously expressed and investigated for their FG degradation ability. The results showed that CotA laccase displayed the highest FG-degrading capacity among the three laccases, achieving 100% FG degradation at 37 °C and pH 7.0 in 1 h without the addition of a redox mediator. Moreover, in vitro and in vivo studies confirmed that the hepatotoxicity of FG was effectively eliminated after oxidative degradation by CotA laccase. Furthermore, the addition of CotA laccase could achieve 87% to 98% FG degradation in defatted CSM within 2 h. In conclusion, CotA laccase can be developed as an effective biocatalyst for the detoxification of FG in CSM.

Enzymatic characterization and application of soybean hull peroxidase as an efficient and renewable biocatalyst for degradation of zearalenone.

Zearalenone (ZEN) is a notorious mycotoxin commonly found in Fusarium-contaminated crops, which causes great loss in livestock farming and serious health problems to humans. In the present work, we found that crude peroxidase extraction from soybean hulls could use H2O2 as a co-substate to oxidize ZEN. Molecular docking and dynamic simulation also supported that ZEN could bind to the active site of soybean hull peroxidase (SHP). Subsequently, SHP extracted from soybean hulls was purified using a combined purification protocol involving ammonium sulfate precipitation, ion exchange chromatography and size exclusion chromatography. The purified SHP showed wide pH resistance and high thermal stability. This peroxidase could degrade 95 % of ZEN in buffer with stepwise addition of 100 µM H2O2 in 1 h. The two main ZEN degradation products were identified as 13-OH-ZEN and 13-OH-ZEN-quinone. Moreover, SHP-catalyzed ZEN degradation products displayed much less cytotoxicity to human liver cells than ZEN. The application of SHP in various food matrices obtained 54 % to 85 % ZEN degradation. The findings in this study will promote the utilization of SHP as a cheap and renewable biocatalyst for degrading ZEN in food.

Improvement of aflatoxin B1 degradation ability by Bacillus licheniformis CotA-laccase Q441A mutant.

Aflatoxin B1 (AFB1) contamination seriously threatens nutritional safety and common health. Bacterial CotA-laccases have great potential to degrade AFB1 without redox mediators. However, CotA-laccases are limited because of the low catalytic activity as the spore-bound nature. The AFB1 degradation ability of CotA-laccase from Bacillus licheniformis ANSB821 has been reported by a previous study in our laboratory. In this study, a Q441A mutant was constructed to enhance the activity of CotA-laccase to degrade AFB1. After the site-directed mutation, the mutant Q441A showed a 1.73-fold higher catalytic efficiency (kcat/Km) towards AFB1 than the wild-type CotA-laccase did. The degradation rate of AFB1 by Q441A mutant was higher than that by wild-type CotA-laccase in the pH range from 5.0 to 9.0. In addition, the thermostability was improved after mutation. Based on the structure analysis of CotA-laccase, the higher catalytic efficiency of Q441A for AFB1 may be due to the smaller steric hindrance of Ala441 than Gln441. This is the first research to enhance the degradation efficiency of AFB1 by CotA-laccase with site-directed mutagenesis. In summary, the mutant Q441A will be a suitable candidate for highly effective detoxification of AFB1 in the future.

Yeast polysaccharide mitigated oxidative injury in broilers induced by mixed mycotoxins via regulating intestinal mucosal oxidative stress and hepatic metabolic enzymes.

This study was aimed to investigate the effects of yeast polysaccharides (YPS) on growth performance, intestinal health, and aflatoxin metabolism in livers of broilers fed diets naturally contaminated with mixed mycotoxins (MYCO). A total of 480 one-day-old Arbor Acre male broilers were randomly allocated into a 2 × 3 factorial arrangement of treatments (8 replicates with 10 birds per replicate) for 6 wk to assess the effects of 3 levels of YPS (0, 1, or 2 g/kg) on the broilers fed diets contaminated with or without MYCO (95 µg/kg aflatoxin B1, 1.5 mg/kg deoxynivalenol, and 490 µg/kg zearalenone). Results showed that mycotoxins contaminated diets led to significant increments in serum malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels, mRNA expressions of TLR4 and 4EBP1 associated with oxidative stress, mRNA expressions of CYP1A1, CYP1A2, CYP2A6, and CYP3A4 associated with hepatic phase Ⅰ metabolizing enzymes, mRNA expressions of p53 associated with hepatic mitochondrial apoptosis, and AFB1 residues in the liver (P < 0.05); meanwhile dietary MYCO decreased the jejunal villus height (VH), villus height/crypt depth (VH/CD), the activity of serum total antioxidant capacity (T-AOC), mRNA expressions of jejunal HIF-1α, HMOX, and XDH associated with oxidative stress, mRNA expressions of jejunal CLDN1, ZO1, and ZO2, and mRNA expression of GST associated with hepatic phase Ⅱ metabolizing enzymes of broilers (P < 0.05). Notably, the adverse effects induced by MYCO on broilers were mitigated by supplementation with YPS. Dietary YPS supplementation reduced the concentrations of serum MDA and 8-OHdG, jejunal CD, mRNA expression of jejunal TLR2, and 4EBP1, hepatic CYP1A2, and p53, and the AFB1 residues in the liver (P < 0.05), and elevated the serum T-AOC and SOD, jejunal VH, and VH/CD, and mRNA expression of jejunal XDH, hepatic GST of broilers (P < 0.05). There were significant interactions between MYCO and YPS levels on the growth performance (BW, ADFI, ADG, and F/G) at d 1 to 21, d 22 to 42, and d 1 to 42, serum GSH-Px activity, and mRNA expression of jejunal CLDN2 and hepatic ras of broilers (P < 0.05). In contrast with MYCO group, the addition of YPS increased BW, ADFI, and ADG, the serum GSH-Px activity (14.31%-46.92%), mRNA levels of jejunal CLDN2 (94.39%-103.02%), decreased F/G, and mRNA levels of hepatic ras (57.83%-63.62%) of broilers (P < 0.05). In conclusion, dietary supplements with YPS protected broilers from mixed mycotoxins toxicities meanwhile keeping normal performance of broilers, presumably via reducing intestinal oxidative stress, protecting intestinal structural integrity, and improving hepatic metabolic enzymes to minimize the AFB1 residue in the liver and enhance the performance of broilers.

Effects of Compound Polysaccharides Derived from Astragalus and Glycyrrhiza on Growth Performance, Meat Quality and Antioxidant Function of Broilers Based on Serum Metabolomics and Cecal Microbiota.

This study aimed to evaluate the effects of dietary supplementation of compound polysaccharides derived from Astragalus and Glycyrrhiza on growth performance, meat quality, antioxidant function, cecal microbiota and serum metabolomics of broilers. A total of 480 one-day-old male Arbor Acres (AA) broilers were randomly divided into four treatments with six replicates comprising 20 broilers each. Treatments: CON group was the basal diet; ANT group was supplemented with Terramycin calcium; LAG group was supplemented with 150 mg/kg Astragalus polysaccharides and 75 mg/kg Glycyrrhiza polysaccharides; HAG group was supplemented with 300 mg/kg Astragalus polysaccharides and 150 mg/kg Glycyrrhiza polysaccharides. The results showed that LAG and HAG supplementation increased growth performance, antioxidant function and meat quality compared with the CON group and ANT group and, especially, the effect of LAG treatment was better than HAG. Analysis of cecal microbiota showed that LAG and HAG supplementation altered cecal microbial diversity and composition in broilers. Serum metabolomics analysis showed that a total of 193 differential metabolites were identified in CON and LAG groups, which were mainly enriched in linoleic acid metabolism and glutathione metabolism pathways. Moreover, there was a close correlation between serum metabolites, cecal microbiota and phenotypic indicators. Conclusion: Dietary supplementation of 150 mg/kg Astragalus polysaccharides and 75 mg/kg Glycyrrhiza polysaccharides could improve the growth performance, antioxidant function and meat quality of broilers by changing the serum metabolites and cecal microbiota composition.

Polysaccharides derived from Astragalus membranaceus and Glycyrrhiza uralensis improve growth performance of broilers by enhancing intestinal health and modulating gut microbiota.

This study was conducted to investigate the effects of dietary supplementation of polysaccharides derived from Astragalus membranaceus and Glycyrrhiza uralensis on growth performance, intestinal health, and gut microbiota composition in broilers. A total of 480 one-day-old male Arbor Acres broilers were randomly divided into 4 treatments with 6 replicates comprising 20 broilers each. Treatments included: basal diet without antibiotics (CON); basal diet supplemented with 500 mg/kg terramycin calcium (ANT); basal diet supplemented with 300 mg/kg Astragalus membranaceus polysaccharides (APS); and basal diet supplemented with 150 mg/kg Glycyrrhiza uralensis polysaccharides (GPS). The results showed that ANT, AP,S and GPS supplementation significantly increased average daily gain (ADG) and decreased feed conversion ratio (FCR) of broilers from 1 to 42 d of age. At 42 d, serum immunoglobulin A (IgA), immunoglobulin M (IgM) and immunoglobulin G (IgG) levels of the APS and GPS group were notably higher than those of the CON group, while serum levels of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) as well as diamine oxidase (DAO) activity in the APS and GPS group were obviously decreased. Moreover, diets supplemented with APS and GPS could significantly increase villus height (VH) and the ratio of villus height to crypt depth (VH/CD) and remarkably upregulated occludin, claudin-1 and mucin-2 (MUC2) mRNA expression in duodenum, jejunum, and ileum of broilers. In addition, 16S rRNA gene sequencing revealed that APS and GPS supplementation altered cecal microbial diversity and composition in broilers. Higher Shannon index was observed in the APS and GPS group compared with the CON group, while GPS supplementation could also increase Chao1 index and Observed species. The result of Principal coordinate analysis (PCoA) showed that microbial community in the CON, ANT, APS, and GPS group clustered separately. Notably, both APS and GPS supplementation significantly decreased the abundance of Bacteroidetes, Bacteroides, Faecalibacterium, Desulfovibrio, and Butyricicoccus, while increased the abundance of Firmicutes, Prevotella, Parabacteroides, Ruminococcus, and Alistipes. The correlation analysis showed that the changes in cecal microbial composition induced by dietary APS and GPS supplementation were closely associated with the alteration of the phenotype of broilers including ADG, FCR, TNF-α, IL-1ß, IL-6, IgA, IgG, DAO, Occludin, Claudin-1, ZO-1, and MUC2. In conclusion, polysaccharides derived from Astragalus membranaceus and Glycyrrhiza uralensis could improve growth performance of broilers by enhancing intestinal health and modulating gut microbiota.

Dietary ellagic acid ameliorated Clostridium perfringens-induced subclinical necrotic enteritis in broilers via regulating inflammation and cecal microbiota.

BACKGROUND: Subclinical necrotic enteritis (SNE), a common intestinal disease of broiler caused by Clostridium perfringens, could reduce production performance of broilers by chronic intestinal damage and poor absorption of nutrients. Ellagic acid (EA) has been reported to present antioxidant and anti-inflammatory properties on human and animals in many aspects. This study was conducted to evaluate the effect and mechanism of EA in relieving SNE in broilers induced by C. perfringens. RESULTS: C. perfringens challenge decreased body weight (BW), average daily gain (ADG), jejunal villi height/crypt depth (V/C) ratio, the activity of catalase (CAT) and the mRNA expression of zonula occludens 1 (ZO-1) in jejunal mucosa of broilers. While feed conversion ratios (FCR), jejunal crypt depth (CD), the activities of myeloperoxidase (MPO) and diamine oxidase (DAO), as well as the concentrations of interleukin 6 (IL-6), C-reactive protein (CRP) and procalcitonin (PCT) in serum, the activities of inducible nitric oxide synthase (iNOS) and lysozyme (LZM), the concentration of malondialdehyde (MDA), and the mRNA expressions of claudin-2, TNF-α, IL-1ß, TLR-4, TLR-2, NF-κB, JAK3, STAT6 and iNOS in jejunal mucosa of broilers were increased by C. perfringens challenge. Dietary EA supplement relieved these adverse effects, and heightened jejunal villi height (VH), the concentration of D-xylose in plasma, activity of superoxide dismutase (SOD), and the mRNA expression of occludin in jejunal mucosa of broilers. The alpha diversity of cecal microbiota indicated that dietary EA supplement increased observed species and Shannon index. C. perfringens challenge increased the relative abundance of Firmicutes and decreased the relative abundance of Desulfobacterota in cecal microbiota. EA increased the relative abundance of Firmicutes in cecal microbiota. LEfSe analysis showed that C. perfringens challenge triggered the imbalance of cecal microbiota in broilers, dietary EA supplementation led to a small beneficial effect on microbiota, while the simultaneous effect of them seemed to stimulate the immune function of broilers by improving the microbiota balance. CONCLUSIONS: Dietary EA ameliorated C. perfringens-induced SNE in broilers via regulating jejunal inflammation signaling pathways TLR/NF-κB and JAK3/STAT6, relieving jejunal oxidative stress and balancing cecal microbiota to inhibit intestinal barrier damage, prevent systemic inflammatory response and improve nutrient absorption capacity, finally protect and enhance growth performance of broilers.

Degradation of zearalenone and aflatoxin B1 by Lac2 from Pleurotus pulmonarius in the presence of mediators.

The contamination of foods and feeds with mycotoxins has been an issue of global significance. For mycotoxin detoxification, enzymatic biodegradation using laccase has received much attention. In this study, a laccase gene lac2 from the fungus Pleurotus pulmonarius was expressed in the Pichia pastoris X33 yeast strain to produce recombinant proteins. Enzymatic properties of recombinant Lac2 and its ability to degrade zearalenone (ZEN) and Aflatoxin B1 (AFB1) in the presence of four mediators (ABTS, TEMPO, AS and SA) were investigated. Result showed that the optimum pH and temperature of recombinant Lac2 were 3.5 and 55 °C, respectively. Lac2 was not sensitive to heat and stable under both acidic and alkaline conditions. Lac2-ABTS and Lac2-AS were efficient systems for ZEN degradation over a wide range of pH (4-8) and temperature (40-60 °C). Lac2-AS was the most efficient system for AFB1 degradation, reaching 99.82% of degradation at pH 7 and 37 °C after 1 h of incubation. Finally, the Lac2-mediator oxidation products were structurally characterized. This study lays a solid foundation for the application of Lac2 laccase combined with AS for degrading mycotoxin in food and feed.

Transcriptional Profiling of Aflatoxin B1-Induced Oxidative Stress and Inflammatory Response in Macrophages.

Aflatoxin B1 (AFB1) is a highly toxic mycotoxin that causes severe suppression of the immune system of humans and animals, as well as enhances reactive oxygen species (ROS) formation, causing oxidative damage. However, the mechanisms underlying the ROS formation and immunotoxicity of AFB1 are poorly understood. This study used the mouse macrophage RAW264.7 cell line and whole-transcriptome sequencing (RNA-Seq) technology to address this knowledge-gap. The results show that AFB1 induced the decrease of cell viability in a dose- and time-dependent manner. AFB1 also significantly increased intracellular productions of ROS and malondialdehyde and decreased glutathione levels. These changes correlated with increased mRNA expression of NOS2, TNF-α and CXCL2 and decreased expression of CD86. In total, 783 differentially expressed genes (DEGs) were identified via RNA-Seq technology. KEGG analysis of the oxidative phosphorylation pathway revealed that mRNA levels of ND1, ND2, ND3, ND4, ND4L, ND5, ND6, Cyt b, COX2, ATPeF0A and ATPeF08 were higher in AFB1-treated cells than control cells, whereas 14 DEGs were downregulated in the AFB1 group. Furthermore, seven immune regulatory pathways mediated by oxidative stress were identified by KEGG analysis. Altogether, these data suggest that AFB1 induces oxidative stress in macrophages via affecting the respiratory chain, which leads to the activation of several signaling pathways related to the inflammatory response.

Novel strategies for degradation of aflatoxins in food and feed: A review.

Aflatoxins are toxic secondary metabolites mainly produced by Aspergillus fungi, posing high carcinogenic potency in humans and animals. Dietary exposure to aflatoxins is a global problem in both developed and developing countries especially where there is poor regulation of their levels in food and feed. Thus, academics have been striving over the decades to develop effective strategies for degrading aflatoxins in food and feed. These strategies are technologically diverse and based on physical, chemical, or biological principles. This review summarizes the recent progress on novel aflatoxin degradation strategies including irradiation, cold plasma, ozone, electrolyzed oxidizing water, organic acids, natural plant extracts, microorganisms and enzymes. A clear understanding of the detoxification efficiency, mechanism of action, degradation products, application potential and current limitations of these methods is presented. In addition, the development and future perspective of nanozymes in aflatoxins degradation are introduced.

CotA laccase, a novel aflatoxin oxidase from Bacillus licheniformis, transforms aflatoxin B1 to aflatoxin Q1 and epi-aflatoxin Q1.

In the present study, the CotA protein from Bacillus licheniformis ANSB821 was cloned and expressed in Escherichia coli. Apart from the laccase activities, we found that the recombinant CotA could effectively oxidize aflatoxin B1 in the absence of redox mediators. The Km, Kcat and Vmax values of the recombinant CotA towards aflatoxin B1 were 60.62 µM, 0.03 s-1 and 10.08 µg min-1 mg-1, respectively. CotA-mediated aflatoxin B1 degradation products were purified and identified to be aflatoxin Q1 and epi-aflatoxin Q1. The treatment of human liver cells L-02 with aflatoxin Q1 and epi-aflatoxin Q1 did not suppress cell viability and induce apoptosis. Molecular docking simulation revealed that hydrogen bonds and van der Waals interaction played an important role in aflatoxin B1-CotA stability. These findings in the current study are promising for a possible application of CotA as a novel aflatoxin oxidase in degrading AFB1 in food.

Enzymatic degradation of deoxynivalenol by a novel bacterium, Pelagibacterium halotolerans ANSP101.

Deoxynivalenol (DON), a toxic secondary metabolite produced by Fusarium species that mainly infests cereals such as wheat and corn, threatens human and livestock health. The present study describes the characterization of a novel bacterial strain, Pelagibacterium halotolerans ANSP101 which is capable of transforming DON to less-toxic product 3-keto-deoxynivalenol by the oxidation of the C3 hydroxyl group. Strain ANSP101 was isolated from a seawater sample from a depth of 55 m in Chinese Bohai sea. The strain was identified as Pelagibacterium halotolerans by morphology characterization and 16S rDNA gene sequencing. The DON degrading activity of strain ANSP101 was predominantly attributed to the bacterial cell lysate. Besides, the cell lysate was sensitive to sodium dodecyl sulfate, heat, and proteinase K treatment, indicating that the intracellular proteins or enzymes are responsible for the DON degradation. The optimal temperature and pH for the maximal degradation of DON were 40 °C and pH 8.0 by the cell lysate. These results provide the potential use of P. halotolerans ANSP101 as a detoxification agent for DON decontamination in cereals and feed.

Characterization and Genome Analysis of a Zearalenone-Degrading Bacillus velezensis Strain ANSB01E.

Zearalenone, a nonsteroidal estrogenic mycotoxin mainly produced by Fusarium species, causes reproductive disorders and hyperestrogenic syndromes in animals and humans. The bacterial strain Bacillus velezensis ANSB01E, isolated from chicken cecal content, was capable of effectively degrading zearalenone in both liquid medium and mouldy corn. Moreover, Bacillus velezensis ANSB01E exhibited good antimicrobial activities against animal pathogenic bacteria, including Escherichia coli, Staphylococcus aureus, and Salmonella spp. Genome-based analysis revealed the presence of genes coding peroxiredoxin and alpha/beta hydrolase in Bacillus velezensis ANSB01E, which may be involved in zearalenone degradation. The study on the genome provides insights into the zearalenone degradation mechanisms and advances the potential application of Bacillus velezensis ANSB01E in food and feed industry.