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      Physicochemical and Antibacterial Properties of Chitosan Extracted from Waste Shrimp Shells

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          Abstract

          This research aims to study the production of chitosan from shrimp shell ( Litopenaeus vannamei) of waste origin using two chemical methodologies involving demineralization, deproteinization, and the degree of deacetylation. The evaluation of the quality of chitosan from waste shrimp shells includes parameters for the yield, physical chemistry characteristics by infrared spectroscopy (FT-IR), the degree of deacetylation, and antibacterial activity. The results showed (by Method 1) extraction yields for chitin of 33% and for chitosan of 49% and a 76% degree of deacetylation. Chitosan obtained by Method 2 was more efficient: chitin (36%) and chitosan (63%), with a high degree of deacetylation (81.7%). The antibacterial activity was tested against Gram-negative bacteria ( Stenotrophomonas maltophilia and Enterobacter cloacae) and Gram-positive Bacillus subtilis and the Minimum Inhibitory Concentrations (MIC) and the Minimum Bactericidal Concentration (MBC) were determined. Method 2 showed that extracted chitosan has good antimicrobial potential against Gram-positive and Gram-negative bacteria and that the process is viable.

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          Preparation and antibacterial activity of chitosan nanoparticles.

          Chitosan nanoparticles, such as those prepared in this study, may exhibit potential antibacterial activity as their unique character. The purpose of this study was to evaluate the in vitro antibacterial activity of chitosan nanoparticles and copper-loaded nanoparticles against various microorganisms. Chitosan nanoparticles were prepared based on the ionic gelation of chitosan with tripolyphosphate anions. Copper ions were adsorbed onto the chitosan nanoparticles mainly by ion-exchange resins and surface chelation to form copper-loaded nanoparticles. The physicochemical properties of the nanoparticles were determined by size and zeta potential analysis, atomic force microscopy (AFM), FTIR analysis, and XRD pattern. The antibacterial activity of chitosan nanoparticles and copper-loaded nanoparticles against E. coli, S. choleraesuis, S. typhimurium, and S. aureus was evaluated by calculation of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Results show that chitosan nanoparticles and copper-loaded nanoparticles could inhibit the growth of various bacteria tested. Their MIC values were less than 0.25 microg/mL, and the MBC values of nanoparticles reached 1 microg/mL. AFM revealed that the exposure of S. choleraesuis to the chitosan nanoparticles led to the disruption of cell membranes and the leakage of cytoplasm.
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            Relationship between antibacterial activity of chitosan and surface characteristics of cell wall.

            Five representative waterborne pathogens were used to illustrate the relationship between chitosan's antibacterial activity and the surface characteristics of the bacterial cell wall. Chitosan was prepared with averaged 75% or 95% deacetylated degree to examine its antibacterial activity against waterborne pathogens. Fresh microbial inoculants for the antibacterial assessment were prepared on nutrient agar at 37 degrees for 24 h. The evaluation items of antibacterial mechanism included hydrophilicity and negative charge analysis of cell surface, and adsorptive characteristics of chitosan to bacterial cell. All the experiments were applied in triplicate tests at least. Although cell wall hydrophilicity was similar among Gram-negative bacteria, the distribution of negative charge on their cell surfaces was quite different. More negatively charged cell surfaces had a greater interaction with chitosan, a phenomenon further confirmed by transmission electron micrography (TEM). Results showed the hydrophilicity in Gram-negative bacteria was much higher than in Gram-positive ones. The correlation coefficient 0.988 between the amount of absorbed chitosan and its inhibition efficiency indicated a close relationship.
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              Applications and Properties of Chitosan

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                Author and article information

                Journal
                Int J Microbiol
                Int J Microbiol
                IJMICRO
                International Journal of Microbiology
                Hindawi Publishing Corporation
                1687-918X
                1687-9198
                2016
                13 July 2016
                : 2016
                : 5127515
                Affiliations
                1Northeast Network for Biotechnology, Federal Rural University of Pernambuco, 52171-900 Recife, PE, Brazil
                2Nucleus of Research in Environmental Sciences and Biotechnology, Catholic University of Pernambuco, 50050-590 Recife, PE, Brazil
                3Doctorate Program in Biological Sciences, Federal University of Pernambuco, 50670-420 Recife, PE, Brazil
                Author notes
                *Galba Maria De Campos-Takaki: galba_takaki@ 123456yahoo.com.br

                Academic Editor: Joseph Falkinham

                Author information
                http://orcid.org/0000-0002-0519-0849
                Article
                10.1155/2016/5127515
                4961830
                27478443
                f4d10c5a-67a1-4370-b6b0-b9c6322b1f55
                Copyright © 2016 José Carlos Vilar Junior et al.

                This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 4 February 2016
                : 3 April 2016
                Funding
                Funded by: Conselho Nacional de Desenvolvimento Científico e Tecnológico
                Funded by: Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco
                Funded by: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
                Categories
                Research Article

                Microbiology & Virology
                Microbiology & Virology

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