Masanori Tohno

Livestock Institute, Lutetia Parisorum, Île-de-France, France

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Publications (53)103.99 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: The radioprotective 105 (RP105)/MD1 complex is a member of the Toll-like receptor (TLR) family. It was reported that RP105/MD1 cooperates with the lipopolysaccharide (LPS) receptor TLR4/MD2 complex and plays a crucial role in the response of immune cells to LPS. This work evaluated whether RP105, TLR4 or TLR2 were involved in the immunoregulatory capacities of Lactobacillus plantarum N14 (LP14) or its exopolysaccharides (EPS). EPS from LP14 were fractionated into neutral (NPS) and acidic (APS) EPS by anion exchange chromatography. Experiments with transfectant HEKRP105/MD1 and HEKTLR2 cells demonstrated that LP14 strongly activated NF-κB via RP105 and TLR2. When we studied the capacity of APS to activate NF-κB pathway in HEKRP105/MD1 and HEKTLR4 cells; we observed that APS strongly stimulated both transfectant cells. Our results also showed that LP14 and APS were able to decrease the production of pro-inflammatory cytokines (IL-6, IL-8 and MCP-1) in porcine intestinal epithelial (PIE) cells in response to enterotoxigenic Escherichia coli (ETEC) challenge. In order to confirm the role of TLR2, TLR4 and RP105 in the immunoregulatory effect of APS from LP14, we used small interfering RNA (siRNA) to knockdown these receptors in PIE cells. The capacity of LP14 and APS to modulate pro-inflammatory cytokine expression was significantly reduced in PIERP105−/− cells. It was also shown that LP14 and APS were capable of upregulating negative regulators of the TLR signaling in PIE cells. This work describes for the first time that a Lactobacillus strain and its EPS reduce inflammation in intestinal epithelial cells in a RP105/MD1-dependend manner.
    Molecular Immunology. 11/2014;
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    ABSTRACT: We investigated concentrations of mycotoxins during the growth of four cultivars of forage maize (Zea mays L.) in Nasushiobara, Tochigi prefecture, and their distribution in ears of maize grown in Morioka, Iwate prefecture, Japan. In experiment 1, we measured concentrations of naturally occurring fumonisin, nivalenol, deoxynivalenol and zearalenone at progressive crop growth stages. Concentrations of fumonisin in stems + leaves remained very low or not detectable, but those in ears became detectable at 40 days after heading and increased rapidly after 50 days after heading (DAH) (fumonisin B1 + B2 <3260 μg/kg; mean value at 50-74 days after heading). Concentrations varied widely within cultivars on the same day. Concentrations of nivalenol, deoxynivalenol and zearalenone in stems + leaves and in ears were low or not detectable throughout the experiment. In experiment 2, we collected three ears of each cultivar at the late yellow-ripe stage that showed extreme symptoms of Fusarium ear rot. Concentrations of fumonisin were extremely high in the upper half of ears in all cultivars (fumonisin B1 + B2 18,000-25,900 μg/kg) but low in the lower half and bracts. Concentrations of nivalenol, deoxynivalenol and zearalenone were extremely low or not detectable. These results show that fumonisin concentrations in ears increased rapidly after 50 DAH, they were extremely high in ears of all cultivars with symptoms of Fusarium ear rot, and fumonisin was the most common contaminant. These results will help reduce mycotoxin contamination.
    Mycotoxin Research 09/2014;
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    ABSTRACT: Immunoregulatory probiotics (immunobiotics) have been proposed to improve piglets' immune system to avoid intestinal infections and reduce unproductive inflammation after weaning. Previously, it was demonstrated that Lactobacillus jensenii TL2937 (LjTL2937) attenuated the inflammatory response triggered by activation of Toll-like receptor 4 (TLR-4) in porcine intestinal epithelial (PIE) cells and antigen presenting cells (APCs) from porcine Peyer's patches (PP).
    BMC Immunology 06/2014; 15(1):24. · 2.61 Impact Factor
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    ABSTRACT: To clarify the changes in rice fumonisin (FUM) concentrations, we conducted field cultivation of 10 forage rice cultivars and inoculation with fumonisin-producing fungal isolates. We cultivated 10 forage rice cultivars at the NARO Institute of Livestock and Grassland Science and one cultivar at two additional farmland sites in Japan in 2011 and 2012. Fusarium fujikuroi, which primarily infects plants shortly after heading, was inoculated on rice just after heading, and we sampled heads at the yellow-ripe and full-ripe stages to assess FUM concentrations. We found differences among cultivars in the FUM concentration and differences among the sites for the same cultivar, but no cultivar had high levels in leaves and stems. Fusarium fujikuroi was the main fumonisin producer. The FUM concentration in heads increases from <1 to 4760 µg/kg DM after the yellow-ripe stage. To control FUM levels, it is necessary to select low-FUM cultivars and manage the cultivation environment.
    Journal of Agricultural and Food Chemistry 03/2014; · 3.11 Impact Factor
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    ABSTRACT: A gram-stain-positive, facultatively anaerobic, non-spore-forming, catalase-negative, coccoid-shaped bacterial strain, designated BV2(T) , was isolated from the vaginal mucosa of a beef cow in Japan. Phylogenetic analysis showed that the isolate had high 16S rRNA gene sequence similarity (92.9%) with Aerococcus suis 1821/02(T) and low similarity (<92.7%) with any other recognized species of the genus Aerococcus. The DNA G + C content was 44.7 mol%, which is within the range observed in Aerococcus species (37.5-48.4 mol%). The major cellular fatty acid was C18:1 ω9c, similar to other Aerococcus type strains. The results of genotypic, phenotypic and chemotaxonomic analyses as well as the low degree of DNA-DNA relatedness with all recognized members of the genus Aerococcus indicate that strain BV2(T) represents a novel Aerococcus species, for which the name Aerococcus vaginalis sp. nov. is proposed. The type strain is BV2(T) (=JCM 19163(T) =DSM 27293(T) ). Emended descriptions of A. sues, A. viridans, A. urinaeequi, A. urinaehominis, A. urinae, A. christensenii and A. sanguinicola are also presented.
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 01/2014; · 2.11 Impact Factor
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    ABSTRACT: We report the 1.86-Mb draft genome and annotation of Lactobacillus oryzae SG293(T) isolated from fermented rice grains. This genome information may provide further insights into the mechanisms underlying the fermentation of rice grains.
    Genome announcements. 01/2014; 2(4).
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    ABSTRACT: Weissella oryzae was originally isolated from fermented rice grains. Here we report the draft genome sequence of the type strain of W. oryzae. This first report on the genomic sequence of this species may help identify the mechanisms underlying bacterial adaptation to the ecological niche of fermented rice grains.
    Genome announcements. 01/2014; 2(4).
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    ABSTRACT: We examined three types of ensiling container and two storage periods with respect to the fermentation quality of rice grain silage, and our findings confirmed that a method previously identified as optimal for silage production at the laboratory scale was also appropriate for farm-scale production. Crushed rice grain was adjusted to 30% moisture content, inoculated with lactic acid bacteria and ensiled in plastic pouches (200 g), plastic drums (15 kg) and flexible containers (100 kg). Samples of silage from the surface layer in the flexible containers had the pH value >5 and fungal growth was observed, but all internal samples had low pH (<4). Extending storage in the drums from 60 to 365 days reduced the lactic acid content of the internal samples, but pH remained at an acceptable level of <4.2. Overall, the production method outlined is appropriate at the farm scale.
    Grassland Science 12/2013; 59(4). · 0.50 Impact Factor
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    ABSTRACT: A Gram-positive, facultatively anaerobic, non-spore-forming and catalase-negative rod-shaped bacterial strain, designated IWT126(T), was isolated from orchardgrass (Dactylis glomerata L.) silage preserved in Hachimantai, Iwate, Japan. The isolate showed growth at 15 to 45 °C, pH 3.5 to 7.5 and with 4.0% (w/v) NaCl. The cell wall peptidoglycan did not contain meso-diaminopimelic acid, and the DNA G+C content was 45.6 mol%. The major cellular fatty acids were C16:0 and C19 cyc 9,10/:1. Based on 16S rRNA gene sequence similarity, strain IWT126(T) was grouped into the genus Lactobacillus and was most closely related to L. odoratitofui YIT 11304(T) (98.7%), L. similis JCM 2765(T) (98.5%), L. collinoides JCM 1123(T) (97.6%), L. paracollinoides DSM 15502(T) (97.6%) and L. kimchicus DCY51(T) (96.9%). Based on sequence analysis of phenylalanyl-tRNA synthase alpha subunit (pheS) gene, strain IWT126(T) was well separated from the phylogenetic neighbors of the genus Lactobacillus. Based on physiological, biochemical and genotypic results, as well as low DNA-DNA relatedness to recognized phylogenetic relatives of the genus Lactobacillus, identification of strain IWT126(T) in the genus Lactobacillus as a novel species named Lactobacillus silagei sp. nov. is proposed. The type strain is IWT126(T) (=JCM 19001(T) =DSM 27022(T).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 08/2013; · 2.11 Impact Factor
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    ABSTRACT: Metabolically active microbiota of the porcine terminal ileum mucosa was analyzed by RT-PCR of 16S rRNAs. The majority of 1951 sequences retrieved (24.8%) displayed the closest similarity to Segmented Filamentous Bacteria (SFB). Phylogenetic analysis inferred the host-specific clusters of SFB sequences suggesting the host-specific selection of this group of bacteria.
    Anaerobe 07/2013; · 2.02 Impact Factor
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    ABSTRACT: Two bacterial strains, designated IWT246(T) and IWT248, were isolated from orchardgrass (Dactylis glomerata L.) silage from Iwate prefecture, Japan, and examined for a taxonomic study. Both organisms were rod-shaped, gram-positive, catalase-negative, facultatively anaerobic, and homofermentative. The cell wall did not contain meso-diaminopimelic acid and the major fatty acids were C18:1ω9c and C19 cyc 9,10/:1. Comparative analyses of 16S rRNA, pheS and rpoA gene sequences revealed that these strains were novel and belonged to the genus Lactobacillus. Based on 16S rRNA gene sequence similarity, the isolates were most closely related to the validated type strains of the genus Lactobacillus: Lactobacillus coryniformis subsp. coryniformis (96.7%), Lactobacillus coryniformis subsp. torquens (96.6%), Lactobacillus bifermentans (95.5%), and Lactobacillus rennini (94.1%). However, the 16S rRNA gene sequences of both IWT246(T) and IWT248 were 99.7% similar to that of the invalid species 'Lactobacillus backii' JCM 18665(T). Genotypic and phenotypic and chemotaxonomic analyses confirmed that these novel strains occupy a unique taxonomic position. DNA-DNA hybridization experiments demonstrated genotypic separation of the novel isolates from related Lactobacillus species. The name Lactobacillus iwatensis sp. nov. is proposed for the novel isolates, with strain IWT246(T) (=JCM 18838(T) =DSM 26942(T)) as the type strain. Our results also suggest that 'Lactobacillus backii' JCM 18665(T) should be classified as a valid novel Lactobacillus species. The cells did not contain meso-diaminopimelic acid in their cell wall peptidoglycan and the major fatty acids were C16:0, C19 cyc 9,10/:1 and summed feature 10. The type strain of this species is JCM 18665(T) (=LMG 23555(T) =DSM 18080(T) =L1062(T)).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 05/2013; · 2.11 Impact Factor
  • Ryuichi Uegaki, Takao Tsukiboshi, Masanori Tohno
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    ABSTRACT: We assessed the production of the mycotoxins fumonisin, deoxynivalenol and zearalenone during the ensiling of corn. Corn was harvested at yellow-ripe or full-ripe stage and separated into the stem and leaf parts and the ear parts, including bracts. Each material was ensiled under five conditions: (1) no fungus added, anaerobic conditions; (2) no fungus added, aerobic conditions; (3) mycotoxin-producing fungus added, anaerobic conditions; (4) mycotoxin-producing fungus added, aerobic conditions; and (5) mycotoxin-producing fungus added to autoclaved material, aerobic conditions. After 40 days of ensilage, we analyzed the silage fermentative quality and mycotoxin concentration. The fermentative quality of all materials was good in treatments (1) and (3), because the pH < 4 increased the lactic acid content preventing mycotoxin levels from increasing. In treatments (2) and (4), fermentative quality of all materials was poor, and mycotoxin levels were slightly increased. In treatment (5), fermentative quality was poor, and mycotoxin levels were increased remarkably. These results indicate that mycotoxins are not produced under anaerobic conditions and are hardly produced under aerobic condition during the ensiling of corn. Our findings suggest that almost all mycotoxins in corn silage are produced pre-harvest.
    Animal Science Journal 04/2013; · 1.04 Impact Factor
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    ABSTRACT: BACKGROUND: Previously, a bovine intestinal epithelial cell line (BIE cells) was successfully established. This work hypothesized that BIE cells are useful in vitro model system for the study of interactions of microbial- or pathogen-associated molecular patterns (MAMPs or PAMPs) with bovine intestinal epithelial cells and for the selection of immunoregulatory lactic acid bacteria (LAB). RESULTS: All toll-like receptor (TLR) genes were expressed in BIE cells, being TLR4 one of the most strongly expressed. We demonstrated that heat-stable PAMPs of enterotoxigenic Escherichia coli (ETEC) significantly enhanced the production of IL-6, IL-8, IL-1alpha and MCP-1 in BIE cells by activating both NF-kappaB and MAPK pathways. We evaluated the capacity of several lactobacilli strains to modulate heat-stable ETEC PAMPs-mediated inflammatory response in BIE cells. Among these strains evaluated, Lactobacillus casei OLL2768 attenuated heat-stable ETEC PAMPs-induced pro-inflammatory response by inhibiting NF-kappaB and p38 signaling pathways in BIE cells. Moreover, L. casei OLL2768 negatively regulated TLR4 signaling in BIE cells by up-regulating Toll interacting protein (Tollip) and B-cell lymphoma 3-encoded protein (Bcl-3). CONCLUSIONS: BIE cells are suitable for the selection of immunoregulatory LAB and for studying the mechanisms involved in the protective activity of immunobiotics against pathogen-induced inflammatory damage. In addition, we showed that L. casei OLL2768 functionally modulate the bovine intestinal epithelium by attenuating heat-stable ETEC PAMPs-induced inflammation. Therefore L. casei OLL2768 is a good candidate for in vivo studying the protective effect of LAB against intestinal inflammatory damage induced by ETEC infection or heat-stable ETEC PAMPs challenge in the bovine host.
    BMC Microbiology 03/2013; 13(1):54. · 2.98 Impact Factor
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    ABSTRACT: The taxonomic position of three Lactobacillus-like microorganisms (strains SG293(T), SG296 and SG310) isolated from fermented rice grain (Oryza sativa L. subsp. japonica) in Japan was investigated. These heterofermentative lactic acid bacteria were Gram-positive, rod-shaped, facultatively anaerobic, non-motile, non-spore-forming, and did not show catalase activity. 16S rRNA gene sequence analysis of strain SG293(T) revealed that the type strains of L. malefermentans (98.3 %), L. odoratitofui (96.2 %), L. similis (96.1 %), L. kimchicus (96.1 %), L. paracollinoides (95.9 %) and L. collinoides (95.7 %) were the closest neighbors. Additional phylogenetic analysis on the basis of pheS and rpoA gene sequences, as well as biochemical and physiological characteristics, indicated that these three strains were members of the genusLactobacillus, and that the novel isolates had a unique taxonomic position. The predominant cellular fatty acids were C18:1 ω9c and C19 cyc 9,10/:1. Because low DNA-DNA hybridization values among the isolates and L. malefermentans JCM 12497(T) were observed, it is proposed that these unidentified isolates be classified as a novel species of the genus Lactobacillus, Lactobacillus oryzae sp. nov. The type strain is SG293(T) (=JCM 18671(T)=DSM 26518(T)).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 02/2013; · 2.11 Impact Factor
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    ABSTRACT: We assessed fumonisin production during the ensiling of rice grain. Rice grain was harvested at the full-ripe stage and prepared as rough rice, crushed rough rice, brown rice or crushed brown rice. Each material was ensiled under six conditions: (1) no fungus, anaerobic; (2) no fungus, aerobic; (3) water added, anaerobic; (4) water and fumonisin-producing fungus added, anaerobic; (5) water and fumonisin-producing fungus added, aerobic; or (6) fumonisin-producing fungus added to autoclaved material, aerobic. After 40 days of ensilage, we analyzed the silage fermentative quality and fumonisin concentration. The fermentative quality of all materials was good in treatments (3) and (4) (pH < 4), reasonable in treatment (5) (pH = 5∼6) and unacceptable in treatments (1) and (2) (pH > 6.5). The fumonisin concentration was low in all materials in treatments (1) to (4), slightly increased in the three materials other than rough rice in treatment (5), and enormously increased in all materials in treatment (6). The results indicate that the fumonisin-producing fungus does not produce fumonisin in anaerobic conditions. It is important that an anaerobic condition be maintained during ensiling in order to reduce the fumonisin content in rice grain silage.
    Animal Science Journal 01/2013; 84(1):48-53. · 1.04 Impact Factor
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    ABSTRACT: To identify the best ensiling process and moisture content for rice grain silage, we determined the effects of additive (i.e. no additive, glucose additive or lactic acid bacteria [LAB] inoculation), crushing process (non‐crushing or crushing) and moisture control (none [13.6%], 20.0, 27.5, 35.0, 42.5 or 50.0% moisture content [in fresh matter, FM]) on rough rice and brown rice. Forage rice harvested at the fully ripe stage was ensiled in small‐scale experiments, and the silage was stored for 60 days. Regardless of the use of additives or LAB inoculation, the pH values tended to decrease with increasing moisture content. LAB inoculation and crushing resulted in low pH values, low butyric acid contents, low ethanol contents and high lactic acid contents, and the lactic acid content increased to more than 2.0% in FM at moisture contents of 27.5% and higher. The parameters of fermentation quality (pH, lactic acid, butyric acid, volatile basic nitrogen and ethanol content) were affected by not only the main effect of additive, crushing process and moisture control but also two‐way or three‐way interaction effects. LAB inoculation and crushing improved the lactic acid fermentation under lower moisture conditions, that is, 27.5% moisture content. In both rough rice and brown rice, we suggest that the appropriate pretreatment is the combined use of LAB inoculation, crushing process and controlling 27.5% moisture content.
    Grassland Science 01/2013; 59(2). · 0.50 Impact Factor
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    ABSTRACT: Metabolically active microbiota of the porcine terminal ileum mucosa was analyzed by RT-PCR of 16S rRNAs. The majority of 1951 sequences retrieved (24.8%) displayed the closest similarity to Segmented Filamentous Bacteria (SFB). Phylogenetic analysis inferred the host-specific clusters of SFB sequences suggesting the host-specific selection of this group of bacteria.
    Anaerobe 01/2013; · 2.02 Impact Factor
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    ABSTRACT: Four strains of Gram-positive, non-spore-forming, rod-shaped, catalase-negative and non-motile lactic acid bacteria, LOOC260(T), LOOC253, LOOC273 and LOOC279, were isolated from timothy grass (Phleum pratense L.) silage produced in Hokkaido, a subarctic region of Japan. These isolates grew at 4-37 °C, indicating the psychrotrophic nature of these strains. Phylogenetic analysis on the basis of 16S rRNA and pheS gene sequences, as well as biochemical and physiological characteristics, indicated that these four strains were members of the genus Lactobacillus. 16S rRNA gene sequence analysis of strain LOOC260(T) demonstrated that the closest neighbours were the type strains of Lactobacillus suebicus (97.7 %), Lactobacillus oligofermentans (96.7 %) and Lactobacillus vaccinostercus (96.7 %). Strain LOOC260(T) showed low levels of DNA-DNA association with L. suebicus JCM 9504(T) (14.7 ± 3.5 %), L. oligofermentans JCM 16175(T) (15.1 ± 4.8 %) and L. vaccinostercus JCM 1716(T) (10.7 ± 3.0 %). The cell wall contained meso-diaminopimelic acid and the major fatty acids were C18:1 ω9c and C19 cyc 9,10/:1. On the basis of phenotypic, physiological and phylogenetic evidence, these isolates represent a novel species of the genus Lactobacillus, for which the name Lactobacillus hokkaidonensis sp. nov. is proposed. The type strain is LOOC260(T) (=JCM 18461(T)=DSM 26202(T)).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 12/2012; · 2.11 Impact Factor
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    ABSTRACT: Paddy rice has been of particular interest as a forage crop in Japan. In this study, the isolated strains TO1000, TO1001, TO1002, and TO1003 were characterized by phenotypic and genotypic approaches. These strains were identified as Lactobacillus plantarum subsp. plantarum by species-specific PCR. Phenotypic characteristics varied among different strains of the same subspecies, and the strains represented unique and diverse phenotypes related to fermentation factors, such as carbohydrate assimilation and range of pH and temperature allowing growth. PCR analysis revealed that the patterns of presence/absence of known plantaricin genes differed in a strain-specific manner. Using these strains as inoculants for preparation of whole crop paddy rice silage, fermentation quality was significantly improved, as shown by lower pH, higher lactic acid content, and inhibition of the growth of undesirable microorganisms such as molds, coliform bacteria, and clostridia, after 30 and 60 days of storage, with effectiveness differing from strain to strain. These observations suggest that suitable candidates for bacterial inoculants in silage preparation should be screened at the strain level. Strain TO1002 may be useful for producing silage inoculants for the production of well-preserved whole crop paddy rice silage.
    FEMS Microbiology Letters 12/2012; 337(2). · 2.05 Impact Factor
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    ABSTRACT: In order to understand the relationship between lactic acid bacteria (LAB) species and silage fermentation, a total of 65 LAB strains isolated from mixed pasture of timothy (Phleum pratense L.) and orchardgrass (Dactylis glomerata L.), and its badly preserved silages were subjected to phenotypic and genetic analysis. According to these analyses, the isolates were divided into 13 groups, including Enterococcus gallinarum, Lactobacillus acidipiscis, L. coryniformis subsp. coryniformis, L. coryniformis subsp. torquens, L. curvatus, L. paraplantarum, L. plantarum subsp. argentoratensis, L. plantarum subsp. plantarum, L. sakei subsp. carnosus, Lactococcus garvieae, Lactococcus lactis subsp. cremoris, Leuconostoc pseudomesenteroides, Pediococcus acidilactici, Pediococcus pentosaceus, Weissella hellenica, Weissella paramesenteroides and Carnobacterium divergens. This is the first report to document that C. divergens, L. acidipiscis, L. sakei subsp. carnosus, L. garvieae, phenotypically novel L. lactis subsp. cremoris, E. gallinarum and W. hellenica are present in vegetative forage crops. L. plantarum group strains were most frequently isolated from the badly preserved silages. Some isolates showed a wide range of growth preferences for carbohydrate utilization, optimal growth pH and temperature in vitro, indicating that they have a high growth potential. These results are useful in understanding the diversity of LAB associated with decayed silage of timothy and orchardgrass.
    Animal Science Journal 04/2012; 83(4):318-30. · 1.04 Impact Factor

Publication Stats

533 Citations
103.99 Total Impact Points

Institutions

  • 2013
    • Livestock Institute
      Lutetia Parisorum, Île-de-France, France
  • 2010–2013
    • National Institute of Livestock and Grassland Science
      Ibaragi, Ōsaka, Japan
  • 2004–2013
    • Tohoku University
      • Graduate School of Agricultural Science
      Sendai, Kagoshima, Japan
  • 2012
    • National Agriculture and Food Research Organization
      Tsukuba, Ibaraki, Japan
  • 2007–2010
    • Shinshu University
      • Graduate School of Agriculture
      Shonai, Nagano, Japan