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      Autochthonous Bacillus licheniformis: Probiotic potential and survival ability in low‐fishmeal extruded pellet aquafeed

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          Abstract

          In recent years, Bacillus spp. have garnered attention as probiotic supplements in aquafeed owing to the production of heat stable and low pH resistant spores. Herein, we isolated and characterized an autochthonous Bacillus licheniformis KCCM 43270 from the intestine of olive flounder ( Paralichthys olivaceus) for supplementation in low‐fishmeal extruded aquafeeds. The KCCM 43270 was screened based on amylase, protease, cellulase, and lipase as well as non‐hemolytic activities. The isolate was able to grow in carboxymethyl cellulose (CMC), xylan, and soybean meal (SBM) when used as a single carbon source in the minimal nutrient M9 medium. The KCCM 43270 spores displayed complete survival in acid (pH 2.5) and bile (0.3%, w/v) for 3 hr, strong biofilm formation, and nearly 50% adhesion with intestinal mucus. The spores of the isolate also showed significant survival ability at 80, 90, 100°C for 60, 30, and 1 min, respectively. In addition, the spores in a blend of SBM complex carrier showed significant heat stability at 120°C for 5 min and under different drying conditions. Furthermore, the spores also survived the extrusion process during low‐fishmeal aquafeed manufacturing, implying the potential application of B. licheniformis KCCM 43270 in aquafeed industry.

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          A rapid and easy method for the detection of microbial cellulases on agar plates using gram's iodine.

          Screening for cellulase-producing microorganisms is routinely done on carboxymethylcellulose (CMC) plates. The culture plates are flooded either with 1% hexadecyltrimethyl ammonium bromide or with 0.1% Congo red followed by 1 M NaCl. In both cases, it takes a minimum of 30 to 40 minutes to obtain the zone of hydrolysis after flooding, and the hydrolyzed area is not sharply discernible. An improved method is reported herein for the detection of extracellular cellulase production by microorganisms by way of plate assay. In this method, CMC plates were flooded with Gram's iodine instead of the reagents just mentioned. Gram's iodine formed a bluish-black complex with cellulose but not with hydrolyzed cellulose, giving a sharp and distinct zone around the cellulase-producing microbial colonies within 3 to 5 minutes. The new method is rapid and efficient; therefore, it can be easily performed for screening large numbers of microbial cultures of both bacteria and fungi. This is the first report on the use of Gram's iodine for the detection of cellulase production by microorganisms using plate assay.
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            Bacillus probiotics: an alternative to antibiotics for livestock production.

            The use of probiotics as feed supplements in animal production has increased considerably over the last decade, particularly since the ban on antibiotic growth promoters in the livestock sector. Several Bacillus sp. are attractive for use as probiotic supplements in animal feed due to their ability to produce spores. Their heat stability and ability to survive the low pH of the gastric barrier represent an advantage over other probiotic micro-organisms. This review discusses important characteristics required for selection of Bacillus probiotic strains and summarizes the beneficial effect of Bacillus-based feed additives on animal production. Although the mechanism of action of Bacillus probiotics has not been fully elucidated, they are effective in improving the growth, survival and health status of terrestrial and aquatic livestock. Bacillus strains also have utility in bioremediation and can reduce nitrogenous waste, thereby improving environmental conditions and water quality. Finally, recent innovative approaches for using Bacillus spores in various applications are discussed.
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              Probiotic characterization of lactic acid bacteria isolated from fermented foods and beverage of Ladakh

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                Author and article information

                Contributors
                sookikim@konkuk.ac.kr
                Journal
                Microbiologyopen
                Microbiologyopen
                10.1002/(ISSN)2045-8827
                MBO3
                MicrobiologyOpen
                John Wiley and Sons Inc. (Hoboken )
                2045-8827
                16 November 2018
                June 2019
                : 8
                : 6 ( doiID: 10.1002/mbo3.2019.8.issue-6 )
                : e00767
                Affiliations
                [ 1 ] Department of Animal Biotechnology Konkuk University Seoul Korea
                [ 2 ] Department of Animal Science and Technology Konkuk University Seoul Korea
                [ 3 ] Department of Marine Biotechnology Gangneung Wonju National University Gangneung Korea
                [ 4 ] Aquafeed Research Center National Institute of Fisheries Science Pohang Korea
                [ 5 ] Department of Marine Life and Applied Sciences Kunsan National University Kunsan Korea
                Author notes
                [*] [* ] Correspondence

                Soo‐Ki Kim, Department of Animal Science and Technology, Konkuk University, Seoul, Republic of Korea.

                Email: sookikim@ 123456konkuk.ac.kr

                [†]

                These authors are the co‐first authors.

                Author information
                http://orcid.org/0000-0003-3499-3330
                Article
                MBO3767
                10.1002/mbo3.767
                6562133
                30444301
                f54ff163-da1c-4f31-ae36-fb5d41eeb935
                © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 03 August 2018
                : 13 October 2018
                : 16 October 2018
                Page count
                Figures: 4, Tables: 1, Pages: 8, Words: 5511
                Funding
                Funded by: National Institute of Fisheries Science, Republic of Korea
                Award ID: R2018019
                Categories
                Original Article
                Original Articles
                Custom metadata
                2.0
                mbo3767
                June 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.6.4 mode:remove_FC converted:13.06.2019

                Microbiology & Virology
                bacillus licheniformis,low‐fishmeal aquafeed,olive flounder,probiotics,survival ability

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