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      Effects of Ensiling Fermentation and Aerobic Deterioration on the Bacterial Community in Italian Ryegrass, Guinea Grass, and Whole-crop Maize Silages Stored at High Moisture Content

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          Abstract

          The effects of storage period and aerobic deterioration on the bacterial community were examined in Italian ryegrass (IR), guinea grass (GG), and whole-crop maize (WM) silages. Direct-cut forages were stored in a laboratory silo for 3, 7, 14, 28, 56, and 120 d without any additives; live counts, content of fermentation products, and characteristics of the bacterial community were determined. 2,3-Butanediol, acetic acid, and lactic acid were the dominant fermentation products in the IR, GG, and WM silages, respectively. The acetic acid content increased as a result of prolonged ensiling, regardless of the type of silage crop, and the changes were distinctively visible from the beginning of GG ensiling. Pantoea agglomerans, Rahnella aquatilis, and Enterobacter sp. were the major bacteria in the IR silage, indicating that alcoholic fermentation may be due to the activity of enterobacteria. Staphylococcus sciuri and Bacillus pumilus were detected when IR silage was spoiled, whereas between aerobically stable and unstable silages, no differences were seen in the bacterial community at silo opening. Lactococcus lactis was a representative bacterium, although acetic acid was the major fermentation product in the GG silage. Lactobacillus plantarum, Lactobacillus brevis, and Morganella morganii were suggested to be associated with the increase in acetic acid due to prolonged storage. Enterobacter cloacae appeared when the GG silage was spoiled. In the WM silage, no distinctive changes due to prolonged ensiling were seen in the bacterial community. Throughout the ensiling, Weissella paramesenteroides, Weissella confusa, and Klebsiella pneumoniae were present in addition to L. plantarum, L. brevis, and L. lactis. Upon deterioration, Acetobacter pasteurianus, Klebsiella variicola, Enterobacter hormaechei, and Bacillus gibsonii were detected. These results demonstrate the diverse bacterial community that evolves during ensiling and aerobic spoilage of IR, GG, and WM silages.

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          Most cited references16

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          Polyphasic study of the spatial distribution of microorganisms in Mexican pozol, a fermented maize dough, demonstrates the need for cultivation-independent methods to investigate traditional fermentations.

          The distribution of microorganisms in pozol balls, a fermented maize dough, was investigated by a polyphasic approach in which we used both culture-dependent and culture-independent methods, including microbial enumeration, fermentation product analysis, quantification of microbial taxa with 16S rRNA-targeted oligonucleotide probes, determination of microbial fingerprints by denaturing gradient gel electrophoresis (DGGE), and 16S ribosomal DNA gene sequencing. Our results demonstrate that DGGE fingerprinting and rRNA quantification should allow workers to precisely and rapidly characterize the microbial assemblage in a spontaneous lactic acid fermented food. Lactic acid bacteria (LAB) accounted for 90 to 97% of the total active microflora; no streptococci were isolated, although members of the genus Streptococcus accounted for 25 to 50% of the microflora. Lactobacillus plantarum and Lactobacillus fermentum, together with members of the genera Leuconostoc and Weissella, were the other dominant organisms. The overall activity was more important at the periphery of a ball, where eucaryotes, enterobacteria, and bacterial exopolysacharide producers developed. Our results also showed that the metabolism of heterofermentative LAB was influenced in situ by the distribution of the LAB in the pozol ball, whereas homolactic fermentation was controlled primarily by sugar limitation. We propose that starch is first degraded by amylases from LAB and that the resulting sugars, together with the lactate produced, allow a secondary flora to develop in the presence of oxygen. Our results strongly suggest that cultivation-independent methods should be used to study traditional fermented foods.
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            Bacterial and fungal communities of wilted Italian ryegrass silage inoculated with and without Lactobacillus rhamnosus or Lactobacillus buchneri.

            To understand the effects of lactic acid bacteria (LAB) inoculation on fermentation products, aerobic stability and microbial communities of silage.
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              DNA-based, culture-independent strategies for evaluating microbial communities in food-associated ecosystems.

              Culture-independent molecular techniques are now available to study microbial ecosystems. They are opening interesting perspectives to problems related to composition and population dynamics of microbial communities in various environmental niches (e.g., soil, water) and foods. In fermented food products, estimates of true microbial diversity is often difficult chiefly on account of the inability to cultivate most of the viable bacteria. The increasing knowledge of gene sequences and the concomitant development of new culture-independent molecular techniques are providing new and effective tools to compare the diversity of microbial communities and to monitor population dynamics in minimally disturbed samples. In this review, recent advances in these techniques are reported. Possible applications to food-associated microbial ecosystems are emphasised.
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                Author and article information

                Journal
                Asian-Australas J Anim Sci
                Asian-australas. J. Anim. Sci
                AJAS
                Asian-Australasian Journal of Animal Sciences
                Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST)
                1011-2367
                1976-5517
                September 2013
                : 26
                : 9
                : 1304-1312
                Affiliations
                College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang 163319, China
                Author notes
                [1]

                Department of Animal Science, Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan.

                [* ] Corresponding Author: Yanbing Li. Tel: +86-459-6819002, Fax: +86-459-6819015, E-mail: yanbing_894@ 123456hotmail.com
                Article
                ajas-26-9-1304-12
                10.5713/ajas.2013.13175
                4093406
                25049913
                9c6b4b9f-8b45-4b7b-9a76-fbe19bb96bbb
                Copyright © 2013 by Asian-Australasian Journal of Animal Sciences

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License http://creativecommons.org/licenses/by-nc/3.0/ which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 27 March 2013
                : 29 April 2013
                : 17 May 2013
                Categories
                Article

                bacteria,denaturing gradient gel electrophoresis,italian ryegrass,guinea grass,whole-crop maize,silage

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