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Two experiments were conducted to investigate the bacterial community of fresh and ensiled paper mulberry prepared with or without lactic acid bacteria (LAB) inoculants in South China. In Experiment 1, the bacterial community, chemical composition, and fermentation products of paper mulberry were analyzed. The results showed that fresh paper mulberry had high crude protein content, buffering capacity value, and amounts of uncultured bacteria. Ensiled paper mulberry showed poor fermentation with high pH value, ammonia-N content, and butyric acid content. In addition, Enterobacter was the dominant genus in silage, followed by Lactobacillus and Enterococcus. Water-soluble carbohydrates, ammonia-N, propionic acid, pH, and lactic acid (LA) were the main factors affecting bacterial community of silage. In Experiment 2, the BP17 (Lactobacillus plantarum) from natural fermented paper mulberry silage and two commercial inoculants (Silage-help [SH] and Chikuso-1 [CH]) were used as additives. Compared with other treatments, BP17 inoculant decreased (p < 0.05) pH and ammonia-N content and increased (p < 0.05) LA content of silage. Inoculation of BP17 also increased the dominance of desirable Lactobacillus and inhibited the growth of harmful bacteria in silage. These results confirmed that paper mulberry could be ensiled and epiphytic LAB inoculant can improve its fermentation quality., (© 2021 Japanese Society of Animal Science.)

To investigate the yeast population dynamics during air exposure in total mixed ration (TMR) silage containing sweet potato residue. TMR were ensiled in laboratory silos (1 kg) with or without two lactic acid bacteria strains, Lactobacillus plantarum (LP), and Lactobacillus amylovorus (LA). Fermentation characteristics were measured and yeast population was investigated by ITS1 region gene sequencing using Illumina MiSeq platform. All treatments were well ensiled, and L. amylovorus improved aerobic stability. During aerobic exposure, Pichia kudriavzevii was detected with increased relative abundance in all treatments and more relative abundant in LP. Pichia fermentans was more relative abundant in control. Higher relative abundance of Pichia anomala was detected in deteriorating LP. The relative abundance of Pichia ohmeri increased during later aerobic exposure in the control and LA, with a significant increase in the count of yeast population. Despite Cryptococcus was detected more relative abundant during early stage of aerobic exposure, the yeast population was below the detection limit. Aerobic deterioration was characterized by an increase in operational taxonomic units of Pichia. High relative abundance of P. anomala and P. kudriavzevii made aerobic deterioration easier. Inhibition of P. fermentans might be an effective strategy for improving the aerobic stability to some instance., (© 2020 Japanese Society of Animal Science.)

This study aimed to investigate the effect of an ensiling period (Experiment 1) and adding lactic acid bacteria (LAB, Experiment 2) on the changes in carotenoid, chlorophyll, and phytol in ensiled Italian ryegrass (IR, Lolium multiflorum Lam.). In Experiment 1, the IR herbage ensiled into plastic bags was analyzed for the contents of photosynthetic pigments and phytol over a 5-week period. During the ensiling process, the β-carotene content decreased (p < .05), whereas the lutein content did not change. Although the chlorophyll content decreased (p < .05) after ensiling, the phytol content barely changed until week 5. In Experiment 2, IR herbage was ensiled without additive, as a Control, or with LAB for 60 days. The pH was lower (p < .05) and lactic acid content was higher (p < .05) for the LAB silage than for the Control. The chlorophyll content in silage was not affected by the LAB; however, the β-carotene content was higher (p < .05) for the LAB silage than for the Control. Phytol and lutein contents in the herbage did not change after ensiling. These results indicate that phytol and lutein in IR herbage can be preserved well in silage, irrespective of their fermentation condition., (© 2019 Japanese Society of Animal Science.)

To effectively utilize crop by-product resources for ruminant feed in semi-arid West Africa, we studied the chemical composition, characteristics concerned with fermentative quality, and microbial population of ensiled pearl millet stover (PMS) and sorghum stover (SS) in Mozambique. After panicle harvest, the PMS and SS were exposed in the field for 7, 21, 35, 49, 63, 77, 91, and 120 days under natural weather conditions. The fresh stover silages were prepared and stored for 120 days. With increased exposure time, the dry matter, neutral detergent fiber, acid detergent lignin, and neutral detergent insoluble nitrogen contents increased, whereas the crude protein, ether extract, gross energy, digestible energy, metabolizable energy, and true protein contents decreased. After 120 days of field exposure, aerobic bacteria dominated both stovers, and lactic acid bacteria (LAB) decreased to below detectable levels. After 120 days of ensiling, LAB dominated the silage of both crops, while the harmful microorganisms as aerobic bacteria, coliform bacteria, yeast, and mold were reduced or below detectable levels. Both silages did not produce more lactic acid to reduce the pH value, but they preserved nutrients well during ensiling. Therefore, PMS and SS can be prepared as silage for ruminant feed in semi-arid West Africa., (© 2020 Japanese Society of Animal Science.)

Enhancing the aerobic stability of silage is very important in silage production. The objective of this study was to compare the roles of a new bacterial strain Lactobacillus parafarraginis ZH1 with Lactobacillus plantarum Chikuso-1 (LP) and Lactobacillus buchneri NCIMB 40788 (LB) in improving the aerobic stability of oat silage and identify the anti-yeast substances produced by them. After ensiled for 45 days, either inoculated silages or control silage were of pH values lower than 4.2. The control and LP inoculated silage had poorer aerobic stability than LB and ZH1 inoculated ones (p < 0.05). ZH1 inoculated silage produced more acetic acid, benzoic acid and hexadecanoic acid than LB inoculated one and had the best aerobic stability (p < 0.05). In MRS medium, strain ZH1 produced more hexadecanoic acid than LP and LB strains, more benzoic acid than strain LP (p < 0.05), and the equal amount of benzoic acid to LB (p > 0.05). In addition, benzoic acid and hexadecanoic acid had low minimal inhibitory concentrations to target yeasts in the pure culture. In conclusion, L. parafarraginis ZH1 had stronger anti-yeast potential and more effectively improved the aerobic stability of silage than other strains used in this study., (© 2018 Japanese Society of Animal Science.)

To investigate the effects of inoculants and environmental temperature on fermentation quality and bacterial diversity of alfalfa silage, first-cut alfalfa was ensiled with or without two screened lactic acid bacteria (LAB) strains, Lactobacillus plantarum, LP, and Lactobacillus casei, LC. Each treatment was divided into three parts and stored at 20°C, 30°C, 40°C, respectively. After 60 days ensiling, fermentation characteristics were measured and bacterial diversity was investigated by 16S ribosomal RNA gene sequencing using Illumina MiSeq platform. LP and LC decreased pH, coliform bacteria counts and increased lactic acid content at 20°C, and the two strains decreased pH, ammonia-N concentration, coliform bacteria counts at 30°C. When the environmental temperature was 40°C, silage treated with LC showed lower LAB and coliform bacteria counts and higher lactic acid content than the untreated and LP treated silages. Butyric acid mainly appeared in silages stored at 40°C. The relative abundance of Lactobacillus in alfalfa silages stored at 20°C and 30°C was highest and increased after LP and LC were added. Garciella was another dominant genus in silages stored at 40°C. In conclusion, LP and LC improved fermentation quality of alfalfa silage by increasing Lactobacillus proportions at 20°C and 30°C; ensiling alfalfa at 40°C was difficult because of Garciella., (© 2018 Japanese Society of Animal Science.)

This study estimated the effects of hybrid and bacterial inoculant on fermentation quality and fatty acid profile of barley silages. Yuyeon (Silkless) and Youngyang (Silking) barley hybrids were harvested at 24.9 and 27.1% dry matter, respectively, and chopped to 10 cm lengths. Each hybrid was treated with or without an inoculant (2 × 10 4  colony-forming units/g of Lactobacillus plantarum). A total of 48 silos were prepared in an experiment with a 2 × 2 (hybrid × inoculant) treatment arrangement with four replications and three ensiling durations (2, 7 and 100 days). After 100 days of ensiling, Yuyeon silage had higher (P < 0.05) in vitro dry matter digestibility and C18:3n-3 than Youngyang silage. Youngyang silage had higher (P < 0.05) acetic acid and C18:2n-6 than Yuyeon silage. Inoculation reduced the C18:3n-3 concentration of both hybrids and increased (P < 0.05) the C18:2n-6 of Youngyang. However, fermentation quality was not improved by the inoculant. These results indicate that Yuyeon hybrid might have better potential benefits on animal performances due to its smooth awn and silkless nature, and higher in vitro dry matter digestibility. Its higher C18:3n-3 would be better for improving fatty acid profile of meat or milk than Youngyang hybrid., (© 2017 Japanese Society of Animal Science.)

In order to assess the survival of lactic acid bacteria (LAB) in whole crop maize silage in the gut of dairy cows, one representative silage sample and three different feces samples were collected from dairy cows on three dairy farms in Hua Bei, China and three dairy farms in Kyushu, Japan. The composition of the bacterial community was examined by denaturing gradient gel electrophoresis and quantitative polymerase chain reaction. Lactobacillus acetotolerans was detected in all bunker-made maize silage samples, regardless of the dairy farm or sampling region from which they were sourced. A total of eight LAB species were detected in the maize silage samples, of which three (L. acetotolerans, L. pontis and L. casei) appeared to survive digestion. The populations of L. acetotolerans in silage and feces were 10 6-7 and 10 3-4 copies/g, respectively, indicating that, even for the LAB species showing potential survival in the gut, competition in this niche may be harsh and the population may substantially decrease during the digestion process. It may be difficult for silage LAB to survive in the gut of silage-fed dairy cows, because marked decrease in population can take place during the digestion process, even for surviving species., (© 2017 Japanese Society of Animal Science.)

The nutritive value and fermentation quality of palisadegrass (Brachiaria brizantha cv. Xaraes) and stylo (Stylosanthes capitata × S. macrocephala cv. Campo Grande) mixed silages were evaluated. The experiment was analyzed in a factorial scheme (5 × 2) in a completely randomized design using increasing levels of stylo (0, 25, 50, 75 and 100% on a fresh matter basis) on palisadegrass silages, with and without microbial inoculants (MI). With the increased ratio of stylo in mixed silages, dry matter (DM), crude protein (CP), acid detergent fiber (ADF), and lignin content increased in silages. The presence of MI promoted lower DM content, and higher neutral detergent fiber corrected for ash and protein, ADF and lignin content. The acid detergent insoluble nitrogen content and the lactic acid bacteria populations were not affected by treatments. The in vitroDM digestibility was affected by the interaction of levels of the stylo and MI. The pH, NH 3 -N/total nitrogen and butyric acid concentrations decreased with increasing levels of stylo. Better nutritive value and quality of fermentation was found in the silage containing higher proportions of this stylo mixed with palisadegrass. The microbial inoculant evaluated did not alter the nutritive value or quality of the fermentation of the silages in this experiment., (© 2017 Japanese Society of Animal Science.)

Four lactic acid bacteria (LAB) strains isolated from straw silages on the Tibetan Plateau were characterized, and their effects on the fermentation quality of Italian ryegrass (Lolium multiflorum Lam.) at different temperatures (10°C, 15°C and 25°C) were studied. These LAB isolates were evaluated using the acids production ability test, morphological observation, Gram staining, physiological, biochemical and acid tolerance tests. All the isolates (M1, LM8, LO7 and LOG9) could grow at 5-20°C, pH 3.5-7.0 and NaCl (3.0%, 6.5%). Strains M1, LM8, LO7 and LOG9 were identified as Lactobacillus plantarum, L. coryniformis, Pediococcus pentosaceus and P. acidilactici, respectively, by sequencing 16S ribosomal DNA. The four isolates were added to Italian ryegrass for ensiling for 30 days at various temperatures. Compared with the corresponding control, inoculating with isolates M1, LM8 and LO7 could improve the silage quality of Italian ryegrass at low temperatures, indicated by significantly (P < 0.05) higher lactic acid (LA) contents and ratios of lactic acid/acetic acid (LA/AA), and significantly (P < 0.05) lower pH and ammonia nitrogen/total nitrogen (AN/TN). Compared with other isolates, LM8 performed better at 10°C and 15°C, indicated by the higher (P < 0.05) LA content and ratio of LA/AA, and the lower (P < 0.05) pH and AN/TN., (© 2017 Japanese Society of Animal Science.)

The objective of this study was to evaluate the effect of grape pomace (GP) with different adding levels (0%, 5%, 10% and 15%, fresh matter basis), alone (GP-LAB) or in combine with an inoculant LAB (GP+LAB), on the fermentation quality and aerobic stability of sweet sorghum silage. After 90 days of ensiling in vacuumized mini-silos, silages were subject to a 7-day aerobic stability test, in which chemical, microbial and polyphenol composition were measured. In the GP-LAB group, adding GP decreased (P < 0.05) concentrations of water-soluble carbohydrate (WSC) and butyric acid in silage. In the GP+LAB group, adding GP increased (P < 0.05) concentrations of lactic acid, WSC and crude protein, decreased (P < 0.05) final pH value, NH 3 -N ratio and butyric acid concentration in silage. Polyphenol level was reduced (P < 0.05) after silage fermentation. During aerobic exposure, the fungi count, pH value and silage temperature increased (P < 0.05), the levels of lactic acid, acetic acid and polyphenols (quercetin 3-O-glucoside and quercetin 3-O-glucuronid) decreased (P < 0.05) in silage. GP+LAB treated silage had a lag phase for aerobic spoilage. When the fermentation products, microbial counts, chemical and polyphenol composition were considered, the use of 10% GP+LAB at ensiling could provide a valuable source for improved fermentation quality and aerobic stability of sweet sorghum silage., (© 2017 Japanese Society of Animal Science.)

Silage is a high-moisture-content forage that is used to feed livestock. Using silage as feed is economically feasible and suitable for cattle management. Secale cereale is considered as a high-quality forage plant. After the heading stage the biomass of S. cereale increases by up to 30%; however, lignification in the cell wall causes low fermentation efficiency and coefficient digestibility, and it may also be contaminated by mycotoxin-producing fungi and can impair the quality of the silage. Therefore, the aim of this study was to screen beneficial Lactobacillius spp. for S. cereale silage to reduce fungi contamination and to increase fibrinolytic ability in the silage. Up to 100 days fermentation period and total 180 lactic acid bacteria were isolated and only a single isolate, R4-26, showed strong acidification ability and antifungal activities against mycotoxin-producing fungus. According to quantitative RT-PCR results, the mycotoxin-producing fungal contamination was significantly reduced in the S. cereale silage containing the R4-26 isolate. In enzymatic assays, only a single isolate, R48-27 demonstrated cellulase, xylanase, chitinase and esterase activities. In conclusion, the beneficial Lactobacillius spp. strains have the ability to reduce mycotoxigenic fungi contamination and increase fibrinolytic activity to improve quality of the S. cereale silage., (© 2017 Japanese Society of Animal Science.)

The effects of pesticides and Lactobacillus plantarum (LP) on fermentation quality of alfalfa (Medicago sativa L.) silage were investigated. Chlorpyrifos and chlorantraniliprole were sprayed on the surface of alfalfa plants at 658.6 and 45.0 g active ingredient/ha, respectively. Alfalfa plants were harvested on day 5 post-application and ensiled with or without LP. Chlorpyrifos and chlorantraniliprole decreased the yeast count of alfalfa material (P < 0.05). Both pesticides increased the butyric acid content of alfalfa silage (P < 0.001). Chlorpyrifos increased pH and decreased lactic acid, acetic acid and short-chain fatty acid contents (P < 0.05). LP decreased pH and butyric acid content, and increased lactic acid and short-chain fatty acid contents of alfalfa silage treated with pesticides (P < 0.05). LP increased the concentration of chlorpyrifos residue in alfalfa silage (P < 0.05). Chlorpyrifos and chlorantraniliprole affected the microbial communities of the material before ensiling, especially coliform bacteria and yeast; the two pesticide residues were reduced after the fermentation of alfalfa silage and affected the fermentation process, whereas LP improved the fermentation quality of pesticides-contaminated alfalfa silage and slowed down the dissipation of chlorpyrifos., (© 2016 Japanese Society of Animal Science.)

Strains TH 14, TH 21 and TH 64 were isolated from tropical silages, namely corn stover, sugar cane top and rice straw, respectively, prepared in Thailand. These strains were selected by low pH growth range and high lactic acid-producing ability, similar to some commercial inoculants. Based on the analysis of 16S ribosomal RNA gene sequence and DNA-DNA relatedness, strain TH 14 was identified as Lactobacillus casei, and strains TH 21 and TH 64 were identified as L. plantarum. Strains TH 14, TH 21, TH 64 and two commercial inoculants, CH (L. plantarum) and SN (L. rhamnosus), were used as additives to fresh and wilted purple Guinea and sorghum silages prepared using a small-scale fermentation method. The number of epiphytic lactic acid bacteria (LAB) in the forages before ensilage was relatively low but the numbers of coliform and aerobic bacteria were higher. Sorghum silages at 30 days of fermentation were all well preserved with low pH (3.56) and high lactic acid production (72.86 g/kg dry matter). Purple Guinea silage inoculated with LAB exhibited reduced count levels of aerobic and coliform bacteria, lower pH, butyric acid and ammonia nitrogen and increased lactic acid concentration, compared with the control. Strain TH 14 more effectively improved lactic acid production compared with inoculants and other strains. © 2016 Japanese Society of Animal Science., (© 2016 Japanese Society of Animal Science.)

In Tibet, it is common practice to make and relocate total mixed ration (TMR) silages before feeding due to the uneven distribution of forages temporally and spatially. This study was conducted to investigate the effects of Lactobacillus plantarum (L), molasses (M) or ethanol (E) on the fermentation quality and aerobic stability of local adaptive TMR silage. After 45 days of ensiling, pH and ammonia nitrogen in inoculated TMR silages were significantly lower than control. During the first 6 days of the aerobic exposure test, a small fluctuation in lactic acid concentration for all TMR silages was observed, and then silages with ethanol continued this trend, while lactic acid in silage without ethanol sharply decreased until the end of the aerobic exposure period. Meanwhile, pH gradually increased along the aerobic exposure; silages treated with ethanol showed lower pH after 9 days of aerobic exposure. The population of yeast gradually increased during 6 days of aerobic exposure, after that an accelerated rise was observed in TMR silages without ethanol. The combinational beneficial effect of L. plantarum and ethanol was found in combined addition of ethanol and Lactobacillus plantarum silages (EL), indicated by intermediate fermentation quality and higher aerobic stability., (© 2015 Japanese Society of Animal Science.)

Lactic acid bacteria (LAB) are suitable for rice straw silage fermentation, but have been studied rarely, and rice straw as raw material for ensiling is difficult because of its disadvantages, such as low nutrition for microbial activities and low abundances of natural populations of LAB. So we investigated the effect of application of LAB and chemical additives on the fermentation quality and microbial community of wilted rice straw silage. Treatment with chemical additives increased the concentrations of crude protein (CP), water soluble carbohydrate (WSC), acetic acid and lactic acid, reduced the concentrations of acid detergent fiber (ADF) and neutral detergent fiber (NDF), but did not effectively inhibit the growth of spoilage organisms. Inoculation with LABs did not improve the nutritional value of the silage because of poor growth of LABs in wilted rice straw. Inoculation with LAB and addition of chemical materials improved the quality of silage similar to the effects of addition of chemical materials alone. Growth of aerobic and facultatively anaerobic bacteria was inhibited by this mixed treatment and the LAB gradually dominated the microbial community. In summary, the fermentation quality of wilted rice straw silage had improved by addition of LAB and chemical materials., (© 2015 Japanese Society of Animal Science.)

To study the microbial population and fermentation dynamics of large needlegrass (LN) and Chinese leymus (CL) during ensiling and subsequent exposure to air, silages were sampled and analyzed using culture-based techniques and denaturing gradient gel electrophoresis (DGGE). A total of 112 lactic acid bacteria (LAB) strains were isolated and identified using the 16S rRNA sequencing method. Lactic acid was not detected in the first 20 days in LN silage and the pH decreased to 6.13 after 45 days of ensiling. The temperature of the LN silage increased after approximately 30 h of air exposure and the CL silage showed a slight temperature variation. Enterococcus spp. were mainly present in LN silage. The proportion of Lactobacillus brevis in CL silage increased after exposure to air. LN silage with a higher proportion of Enterococcus spp. and propionic acid concentration did not show higher fermentation quality or aerobic stability than CL silage, which had a higher concentration of acetic acid, butyric acid and increased proportion of L. brevis after exposure to air., (© 2015 Japanese Society of Animal Science.)

In order to effectively use the cassava (Manihot esculenta Crantz) residues, including cassava leaves, peel and pulp for livestock diets, the chemical and microbiological composition, silage preparation and the effects of lactic acid bacteria (LAB) inoculants on silage fermentation of cassava residues were studied. These residues contained 10(4) to 10(5) LAB and yeasts, 10(3) to 10(4) coliform bacteria and 10(4) aerobic bacteria in colony forming units (cfu) on a fresh matter (FM) basis. The molds were consistently at or below the detectable level (10(2) cfu of FM) in three kinds of cassava residues. Dry matter (DM), crude protein (CP) and neutral detergent fiber (NDF) content of cassava residues were 17.50-30.95%, 1.30-16.41% and 25.40-52.90% on a DM basis, respectively. The silage treatments were designed as control silage without additive (CO) or with LAB inoculants Chikuso-1 (CH, Lactobacillus plantarum) and Snow Lacto (SN, Lactobacillus rhamnosus) at a rate of 5 mg/kg of FM basis. All silages were well preserved with a low pH (below 4.0) value and when cassava residues silage treated with inoculants CH and SN improved fermentation quality with a lower pH, butyric acid and higher lactic acid than control silage., (© 2015 Japanese Society of Animal Science.)

The objective of this experiment was to evaluate the effects of replacement of whole-plant corn with oat and common vetch on the fermentation quality, chemical composition and aerobic stability of total mixed ration (TMR) silage in Tibet. Four TMR that varied in the forage sources on dry matter basis were used: (i) 52% whole-plant corn (Control); (ii) 43% oat + 12% common vetch (OC3.6); (iii) 38% oat + 18% common vetch (OC2.2); and (iv) 33% oat + 23% common vetch (OC1.5). Silos were opened on day 45 and then subjected to an aerobic stability test for 12 days. The results showed that all silages were well preserved with low pH and NH3 -N, and high lactic acid and V-scores. With the increasing proportion of common vetch, crude protein, ether extract increased (P < 0.05), and neutral detergent fiber and acid detergent fiber decreased (P < 0.05). Under aerobic conditions, treated silages were more stable than the control silage as indicated by lower (P < 0.05) pH and yeast population. It was concluded that replacement of whole-plant corn with oat and common vetch had no unfavorable effects on the fermentation quality and improved crude protein content and aerobic stability of TMR silage. OC2.2 silage was the best among three treated TMR silages., (© 2014 Japanese Society of Animal Science.)

As a forage source for total mixed ration (TMR) silage production, locally produced crop silage is now used in addition to imported hay. This type of TMR ensiling is regarded as a two-step fermentation process; hence, a survey was carried out to determine whether the bacteria in crop silage affect the subsequent TMR ensiling. Fermentation product contents and bacterial community were determined for TMR silage and its ingredient silages collected in August, October and November. August product contained corn, sorghum and Italian ryegrass silages, October product had wheat silage exclusively and November product did not include any crop silages. Acetic acid, lactic acid, 2,3-butanediol and ethanol were predominant fermentation products in corn, sorghum, Italian ryegrass and wheat silages, respectively. Robust lactic acid fermentation was seen in TMR silage, even if acetate-type and alcohol-type silages were mixed as ingredients. The finding that bacterial community of the TMR silage appeared unrelated to those of ingredient silage supported this. Silages of various fermentation types can therefore be formulated without interfering with lactate-type fermentation in TMR silage., (© 2014 Japanese Society of Animal Science.)

To evaluate the current practice of corn silage management in China, samples of bunker-made silage were collected from 14 farms within a 500-km radius of Beijing for the analysis of fermentation products and bacterial communities. Mean values for dry matter (DM) content were as low as 250 g/kg in both corn stover (St) and whole crop corn (Wc) silages, and pH values averaged 4.48 and 3.73, respectively. Only three of the 14 silages exhibited a lactic-to-acetic acid ratio > 1.0, indicating that the presence of acetic acid was predominant in fermentation. Although 1,2-propanediol content was marginal in most cases (< 5.0 g/kg dry matter (DM)), two Wc silages had 1,2-propanediol levels > 25 g/kg DM. In contrast, 3 St silages had large amounts (> 10 g/kg DM) of butyric acid, and two of the three butyrate silages also had high concentrations of 1-propanol. Denaturing gradient gel electrophoresis analysis demonstrated that the bacterial community appeared similar in 10 out of the 14 silage samples. Bands indicating Lactobacillus buchneri, L. acetotolerans and Acetobacter pasteurianus were found in both the St and Wc silages, accounting for the high acetic acid content found across silage samples., (© 2013 Japanese Society of Animal Science.)

To gain further insights into temperate and tropical grass ensiling, fermentation products and bacterial communities were examined at both the initial and late stages of ensiling of wilted Italian ryegrass and wilted guinea grass silages. 2,3-Butanediol and ethanol fermentation were observed in wilted Italian ryegrass silage. Enterobacteria such as Rahnella sp. and Enterobacter sp. may have been involved in fermentation; however, alcohol production was intensified after the silage enterobacterial community overwhelmed the pre-ensiled enterobacterial community. Pediococcus spp. appeared in silage stored for 4 months, when a significant increase in lactic acid content was seen compared with that at 2 months. Prolonged storage enhanced acetic acid fermentation in wilted guinea grass silage. The disappearance of Enterococcus sulfureus and appearance of Lactobacillus plantarum may have been associated with the increased acetic acid content. Although many species of enterobacteria were found in common between the pre-ensiled crop and silages of Italian ryegrass and guinea grass, marked differences were seen in the type of fermentation from the initial stages. These results indicate that the bacterial community of pre-ensiled crops may be immediately replaced by one that is adapted to ensiling environments, although metabolic changes may continue over the course of ensiling., (© 2013 Japanese Society of Animal Science.)

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., (© 2012 The Authors. Animal Science Journal © 2012 Japanese Society of Animal Science.)

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., (© 2011 The Authors. Animal Science Journal © 2011 Japanese Society of Animal Science.)

Twenty-three lactic acid bacteria (LAB) isolated from three cultivars (Akiaoba, Nagahahikari and Tachiwase) of Italian ryegrass (Lolium multiflorum Lam.) silage were precisely characterized by a combination of phenotypic tests, genotypic 16S ribosomal DNA sequencing and rapid PCR-based analyses, focusing on their useful phenotypes for silage preparation as inoculants. We successfully identified both at the species and subspecies levels: phenotypically novel Lactococcus lactis subsp. lactis, Lactobacillus brevis, Lactobacillus coryniformis subsp. torquens, Lactobacillus curvatus, Lactobacillus plantarum subsp. plantarum, Lactobacillus sakei subsp. carnosus, Leuconostoc mesenteroides subsp. dextranicum and Pediococcus parvulus. This is the first report to elucidate the presence of Lactobacillus coryniformis ssp. torquens and Leuconostoc mesenteroides subsp. dextranicum in Italian ryegrass silages. Physiological and biochemical tests revealed that phenotypic characteristics are different among the different strains of the same species and subspecies, and that the isolates show unique and diverse phenotypes related to fermentation factors, such as available carbohydrates, optimal growth pH and temperature. These results suggest that, for various well-preserved silage preparations, the isolates may be useful in producing novel inoculants corresponding to their optimally climatic and ecological niches., (© 2011 The Authors. Animal Science Journal © 2011 Japanese Society of Animal Science.)

Two lactic acid bacteria (LAB) strains, Pediococcus pentosaceus SC1 and Lactobacillus paraplantarum SC2 isolated from king grass silage, were characterized and their effectiveness to improve the silage fermentation quality of stylo (Stylosanthes guianensis Sw.) was studied. Strain SC1 was able to grow at a high temperature of 45°C, while SC2 did not. SC2 normally grew at a low pH of 4.0, while SC1 could not. These two strains and a commercial inoculant of LAB (L. plantarum, LP) were used as additives to stylo silage preparation at various temperatures (20°C, 30°C and 40°C). All LAB inoculants significantly (P < 0.05) reduced the pH value and ammonia-N content, and increased the ratio of lactic acid to acetic acid and quality score compared with the control. In addition, inoculating LAB strains markedly (P < 0.05) reduced butyric acid content at the temperatures of 30°C and 40°C. Compared to SC2 and LP strains, strain SC1 was the most effective for improving stylo silage quality at 20°C, indicated by the increase in lactic acid, ratio of lactic acid to acetic acid and quality score. At 30°C and 40°C, there were no significant differences among SC1, SC2 and LP treatments in pH values, contents of acetic acid, butyric acid and ammonia-N (P > 0.05)., (© 2011 The Authors. Animal Science Journal © 2011 Japanese Society of Animal Science.)

One hundred and twenty-six strains were isolated from corn stover in Henan Province, China, of which 105 isolates were considered to be lactic acid bacteria (LAB) according to Gram-positive, catalase-negative and mainly metabolic lactic acid product. Analysis of the 16S ribosomal DNA sequence of 21 representative strains was used to confirm the presence of the predominant groups and to determine the phylogenetic affiliation of isolates. The sequences from the various LAB isolates showed high degrees of similarity to those of the GenBank type strains between 99.4% and 100%. The prevalent LAB, predominantly Lactobacillus (85.6%), consisted of L. plantarum (33.3%), L. pentosus (28.6%) and L. brevis (23.7%). Other LAB species as Leuconostoc lactis (4.8%), Weissella cibaria (4.8%) and Enterococcus mundtii (4.8%) also presented in corn stover. The present study is the first to fully document corn stover-associated LAB involved in the silage fermentation. The identification results revealed LAB composition inhabiting corn stover and enabling the future design of appropriate inoculants aimed at improving the fermentation quality of silage., (© 2011 The Authors. Animal Science Journal © 2011 Japanese Society of Animal Science.)

In order to clarify the ensiling characteristics of stylo (Stylosanthes guianensis Swartz), the effects of wilting (no wilting, light wilting and heavy wilting) and storage temperatures (10°C, 20°C, 30°C and 40°C) on the fermentation quality and aerobic stability of stylo silage were investigated. Wilting had no significant influence on the contents of crude protein, ether extract and acid detergent fiber, and numbers of lactic acid bacteria, aerobic bacteria, yeasts and mold (P > 0.05). Heavy wilted material, wilted for 12 h, had higher neutral detergent fiber content and lower water-soluble carbohydrate content than unwilted and light wilted materials (P < 0.05). Wilting and storage temperatures had significant effects on pH value, acetic acid, butyric acid and NH(3) -N contents of stylo silage (P < 0.01 or P < 0.05). Wilting tended to reduce acetic acid and NH(3) -N contents and improve the fermentation quality of stylo silage. In all the silages, no wilting silage ensiled at 30°C had the highest butyric acid content (P < 0.05). High temperature of 40°C markedly restricted the growth of lactic acid bacteria and aerobic bacteria in silage, irrespective of wilting. The wilted silage or silage stored at low temperature had poor aerobic stability., (2011 The Authors. Animal Science Journal © 2011 Japanese Society of Animal Science.)

Total mixed ration (TMR) silage in different seasons was prepared with apple refuse, orchardgrass hay, orchardgrass silage, corn silage and a commercial compound feed in Tohoku, Japan, and with tofu cake, brewers' grains, tall fescue hay, Sudangrass hay, timothy hay, alfalfa hay and a commercial compound feed in Shikoku, Japan, respectively, and their fermentation quality and microorganism composition were evaluated. In Tohoku, the TMR silage in spring, autumn and winter displayed poor quality, with pH value above 4.7, and the harmful microorganisms such as escherichia, yeasts and molds were detected even after 60 days of ensiling; but the TMR silage in summer was well preserved and the escherichia were too few to count after 7 days of ensiling. In Shikoku, the TMR silage in all seasons was well preserved, with low pH, high lactic acid contents, and escherichia, yeasts and molds were consistently at or below the detectable numbers after 7 days of ensiling. The results showed that the TMR silage could not produce sufficient lactic acid to improve silage quality under low-temperature conditions. Therefore, it is necessary to develop preparation techniques of promoting the lactic acid fermentation for TMR silage in cold seasons., (© 2011 The Authors. Journal compilation © 2011 Japanese Society of Animal Science.)