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      Aflatoxin B 1 Degradation and Detoxification by Escherichia coli CG1061 Isolated From Chicken Cecum

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          Abstract

          Aflatoxin B 1 (AFB 1) is one of the most hazardous mycotoxins contamination in food and feed products, which leads to hepatocellular carcinoma in humans and animals. In the present study, we isolated and characterized an AFB 1 degrading bacteria CG1061 from chicken cecum, exhibited an 93.7% AFB 1 degradation rate by HPLC. 16S rRNA gene sequence analysis and a multiplex PCR experiment demonstrated that CG1061 was a non-pathogenic Escherichia coli. The culture supernatant of E. coli CG1061 showed an 61.8% degradation rate, whereas the degradation rates produced by the intracellular extracts was only 17.6%, indicating that the active component was constitutively secreted into the extracellular space. The degradation rate decreased from 61.8 to 37.5% when the culture supernatant was treated with 1 mg/mL proteinase K, and remained 51.3% when that treated with 100°C for 20 min. We postulated that AFB 1 degradation was mediated by heat-resistant proteins. The content of AFB 1 decreased rapidly when it was incubated with the culture supernatant during the first 24 h. The optimal incubation pH and temperature were pH 8.5 and 55°C respectively. According to the UPLC Q-TOF MS analysis, AFB 1 was bio-transformed to the product C 16H 14O 5 and other metabolites. Based on the results of in vitro experiments on chicken hepatocellular carcinoma (LMH) cells and in vivo experiments on mice, we confirmed that CG1061-degraded AFB 1 are less toxic than the standard AFB 1. E. coli CG1061 isolated from healthy chicken cerum is more likely to colonize the animal gut, which might be an excellent candidate for the detoxification of AFB 1 in food and feed industry.

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          Most cited references35

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          Diarrheagenic Escherichia coli.

          Escherichia coli is the predominant nonpathogenic facultative flora of the human intestine. Some E. coli strains, however, have developed the ability to cause disease of the gastrointestinal, urinary, or central nervous system in even the most robust human hosts. Diarrheagenic strains of E. coli can be divided into at least six different categories with corresponding distinct pathogenic schemes. Taken together, these organisms probably represent the most common cause of pediatric diarrhea worldwide. Several distinct clinical syndromes accompany infection with diarrheagenic E. coli categories, including traveler's diarrhea (enterotoxigenic E. coli), hemorrhagic colitis and hemolytic-uremic syndrome (enterohemorrhagic E. coli), persistent diarrhea (enteroaggregative E. coli), and watery diarrhea of infants (entero-pathogenic E. coli). This review discusses the current level of understanding of the pathogenesis of the diarrheagenic E. coli strains and describes how their pathogenic schemes underlie the clinical manifestations, diagnostic approach, and epidemiologic investigation of these important pathogens.
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            Degradation of aflatoxin B(1) by fungal laccase enzymes.

            The enzymatic degradation of aflatoxin B(1) (AFB(1)) by white rot fungi through laccase production was investigated in different liquid media. A significant (P<0.0001) correlation was observed between laccase activity and AFB(1) degradation exhibited by representatives of Peniophora and Pleurotus ostreatus cultivated in minimal salts (MSM) (r=0.93) and mineral salts - malt extract (MSB-MEB) (r=0.77) liquid media. Peniophora sp. SCC0152 cultured in MSB-MEB liquid medium supplemented with veratryl alcohol and sugarcane bagasse showed high laccase activity (496U/L), as well as 40.45% AFB(1) degradation as monitored using high performance liquid chromatography. P.ostreatus St2-3 cultivated in MSM liquid medium supplemented with veratryl alcohol resulted in laccase activity of 416.39U/L and 35.90% degradation of AFB(1). Aflatoxin B(1) was significantly (P<0.0001) degraded when treated with pure laccase enzyme from Trametes versicolor (1U/ml, 87.34%) and recombinant laccase produced by Aspergillus niger D15-Lcc2#3 (118U/L, 55%). Aflatoxin B(1) degradation by laccase enzyme from T. versicolor and recombinant laccase enzyme produced by A. niger D15-Lcc2#3 coincided with significant (P<0.001) loss of mutagenicity of AFB(1), as evaluated in the Salmonella typhimurium mutagenicity assay. The degradation of AFB(1) by white rot fungi could be an important bio-control measure to reduce the level of this mycotoxin in food commodities.
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              Biological degradation of aflatoxin B1 by Rhodococcus erythropolis cultures.

              Aflatoxin contamination of food and grain poses a serious economic and health problem worldwide, but particularly in Africa. Aflatoxin B(1) (AFB(1)) is extremely mutagenic, toxic and a potent carcinogen to both humans and livestock and chronic exposure to low levels of AFB(1) is a concern. In this study, the biodegradation of aflatoxin B(1) (AFB(1)) by Rhodococcus erythropolis was examined in liquid cultures using thin layer chromatography (TLC), high performance liquid chromatography (HPLC), electro spray mass spectrometry (ESMS) and liquid chromatography mass spectrometry (LCMS). AFB(1) was effectively degraded by extracellular extracts from R. erythropolis liquid cultures. Results indicated that the degradation is enzymatic and that the enzymes responsible for the degradation of AFB(1) are extracellular and constitutively produced. Furthermore, the biodegradation of AFB(1) when treated with R. erythropolis extracellular fraction coincided with a loss of mutagenicity, as evaluated by the Ames test for mutagenicity.
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                Author and article information

                Contributors
                Journal
                Front Pharmacol
                Front Pharmacol
                Front. Pharmacol.
                Frontiers in Pharmacology
                Frontiers Media S.A.
                1663-9812
                17 January 2019
                2018
                : 9
                : 1548
                Affiliations
                [1] 1Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University , Guangzhou, China
                [2] 2Key Laboratory of Zoonosis of Ministry of Agriculture and Rural Affairs, South China Agricultural University , Guangzhou, China
                [3] 3South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences , Guangzhou, China
                Author notes

                Edited by: Hani El-Nezami, The University of Hong Kong, Hong Kong

                Reviewed by: Carol Verheecke-Vaessen, Cranfield University, United Kingdom; Sascha Rohn, Universität Hamburg, Germany; Silvia Gratz, University of Aberdeen, United Kingdom

                *Correspondence: Yiqun Deng, yqdeng@ 123456scau.edu.cn

                These authors have contributed equally to this work

                This article was submitted to Predictive Toxicology, a section of the journal Frontiers in Pharmacology

                Article
                10.3389/fphar.2018.01548
                6344451
                30705630
                52457268-c250-4644-b750-5d09cac42013
                Copyright © 2019 Wang, Wu, Liu, Shi, Liu, Xu, Hao, Mu, Deng and Deng.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 30 September 2018
                : 18 December 2018
                Page count
                Figures: 5, Tables: 2, Equations: 0, References: 43, Pages: 9, Words: 0
                Categories
                Pharmacology
                Original Research

                Pharmacology & Pharmaceutical medicine
                aflatoxin b1,escherichia coli,degradation,detoxification,chicken cecum

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