Faculty Profiles - NISHINO Naoki

Papers - NISHINO Naoki

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Effects of inoculation with lactic acid bacteria on the bacterial communities of Italian ryegrass, whole crop maize, guinea grass and rhodes grass silages

S. Parvin, C. Wang, Y. Li, N. Nishino

ANIMAL FEED SCIENCE AND TECHNOLOGY 160 ( 3-4 ) 160 - 166 2010.9

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

Italian ryegrass (IR), whole crop maize (WM), guinea grass (GG) and rhodes grass (RG) were stored in laboratory silos for 30 d with and without Lactobacillus plantarum or Lactobacillus brevis inoculation. The grasses were wilted to low moisture levels (about 500 g/kg) prior to ensiling, while WM was ensiled directly after harvest. Microbial counts and bacterial community analyses were undertaken by plate-culture and denaturing gradient gel electrophoresis (DGGE), respectively, on pre- and post-ensiled crops. A total of 31 DGGE bands were retrieved, cloned, and sequenced to identify bacterial species according to their 16S rRNA gene sequences. Inoculation with L plantarum and L brevis increased (P<0.05) lactic acid and decreased (P<0.05) 2,3-butanediol contents in wilted grass silages, with higher (P<0.05) contents of lactic acid observed in L plantarum-inoculated silages compared to L brevis-inoculated silages. Inoculation with L. brevis increased (P<0.05) both lactic and acetic acid contents in IR silage and intensified lactic acid production in GG and RG silages. Inoculation with either L plantarum or L brevis did not affect fermentation of WM silage, except that the lactic acid to acetic acid ratio increased (P<0.05) in L plantarum-inoculated silage. Differences in inoculation effects could be accounted for by changes in bacterial communities present in the silages. Interestingly, in wilted grass silages, the inoculation nearly eliminated the indigenous communities, leaving the inoculated species as the principal bacteria. In maize silage, desirable bacterial communities were found in untreated control, and the inoculated lactic acid bacteria (LAB) were additive to the uninoculated communities. Combination of fermentation product analysis and bacterial community analysis may describe the effects of LAB inoculation on the ensiling process. (C) 2010 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.anifeedsci.2010.07.010

Succession of lactic acid bacteria in wilted rhodesgrass silage assessed by plate culture and denaturing gradient gel electrophoresis Reviewed

Suraya Parvin, Naoki Nishino

GRASSLAND SCIENCE 56 ( 1 ) 51 - 55 2010.3

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-BLACKWELL PUBLISHING, INC

Lactic acid bacteria (LAB) in low-moisture rhodesgrass (Chrolis gayana Kunth) silage, involved in the fermentation process, were determined by plate culture and denaturing gradient gel electrophoresis (DGGE). The first harvest of rhodesgrass was wilted (581 g dry matter kg-1) and stored in laboratory silos for 15, 30, 90 and 180 days. The concentration of acetic acid was found to be more than that of lactic acid until day 30, while lactic acid was the major fermentation product after day 90. Both plate culture and DGGE analyses indicated that Lactobacillus plantarum, Lactobacillus brevis and Lactococcus lactis were the principal LAB present during the period of ensiling. Similar results were obtained with the two methods with respect to the LAB species, but neither of the methods could account for the change in the fermentation products over the period of ensiling.

DOI： 10.1111/j.1744-697X.2009.00173.x

Bacterial community associated with ensilage process of wilted guinea grass Reviewed

S. Parvin, N. Nishino

JOURNAL OF APPLIED MICROBIOLOGY 107 ( 6 ) 2029 - 2036 2009.12

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-BLACKWELL PUBLISHING, INC

Aims:
To determine the effects of wilting, storage period and bacterial inoculant on the bacterial community and ensiling fermentation of guinea grass silage.
Methods and Results:
Fermentation products, colony counts and denaturing gradient gel electrophoresis (DGGE) profiles were determined. There was more lactic acid than acetic acid in all silages, but the lactic acid to acetic acid ratio decreased with storage time. This shift from lactic to acetic acid was not prevented even with a combination of wilting and bacterial inoculant. The DGGE analyses suggest that facultatively heterofermentative lactic acid bacteria (Lactobacillus plantarum, Lactobacillus brevis and Lactobacillus pentosus) were involved in the shift to acetic acid fermentation.
Conclusions:
Lactic acid can dominate the fermentation in tropical grass silage with sufficient wilting prior to ensiling. Prolonged storage may lead to high levels of acetic acid without distinctive changes in the bacterial community.
Significance and Impact of the Study:
The bacterial community looks stable compared to fermentation products over the course of long storage periods in tropical grass silage. Acetic acid fermentation in tropical grass silage can be a result of the changes in bacterial metabolism rather than community structure.

DOI： 10.1111/j.1365-2672.2009.04391.x

Association of Lactobacillus buchneri with aerobic stability of total mixed ration containing wet brewers grains preserved as a silage Reviewed

Fujin Wang, Naoki Nishino

ANIMAL FEED SCIENCE AND TECHNOLOGY 149 ( 3-4 ) 265 - 274 2009.3

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

Laboratory-scale plastic silos were prepared to examine bacterial factors associated with the aerobic stability of total mixed rations (TMR) preserved as a silage after mixing, In experiment 1, TMR silages containing wet brewers grains (BG), hay (lucerne or sudan-grass), maize grain, wheat bran, beet pulp and liquid molasses, were ensiled for 14 and 56 days, and bacterial communities were examined by denaturing gradient gel electrophoresis (DGGE). The silages resisted aerobic deterioration for as long as 2 weeks, except that heating occurred after 5.5 days in 14-day silage prepared with sudangrass hay. The DGGE analyses indicated that, although not found in the materials, Lactobacillus buchneri and L. brevis were detectable in long-stored TMR silage, L delbriteckii and L. fermentum were found in the materials, and appeared to survive during fermentation and after exposure to air. There were few changes in the DGGE profiles before and after heating in deteriorated silage containing sudangrass hay. In experiment 2, TMR silage with ingredients similar to those in experiment I was prepared, and culturable lactic acid bacteria were isolated from the plates of de Man-Rogosa-Sharpe agar. L delbrueckii and L. fermentum were obtained from the materials but not from TMR silage. L buchneri and L brevis were not found in the materials, but more than one-third of the isolates were L buchneri and L. brevis in 14- and 56-day silages, respectively. In experiment 3, whole crop maize was inoculated with L. buchneri, L brevis, L delbrueckii and L. fermentum isolates, and fermentation and aerobic stability were determined. No changes occurred from L. brevis, L delbrueckii and L fermentum inoculation, while aerobic spoilage was suppressed by L buchneri with increases in pH and acetic acid content. Results indicate that, whilst not found in BG or the TMR mixture, L buchneri can become detectable during fermentation, and this bacterium helps establish aerobic stability of BG-containing TMR silage. (c) 2008 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.anifeedsci.2008.06.012

Ensiling of soybean curd residue and wet brewers grains with or without other feeds as a total mixed ration Reviewed

F. Wang, N. Nishino

Journal of Dairy Science 91 ( 6 ) 2380 - 2387 2008.6

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：AMER DAIRY SCIENCE ASSOC

Wet brewers grains and soybean curd residue were stored in laboratory-scale silos without (BG and SC silages, respectively) or with other ingredients as total mixed rations (BGT and SCT silages, respectively). Silages were opened after 14 and 56 d, and microbial counts, fermentation products, and aerobic stability were determined. Denaturing gradient gel electrophoresis was carried out to examine bacterial communities, and several bacteria that appeared to be involved in fermentation were identified. Lactic acid content was greater in SCT than in BGT silage, but lower in SC than in BG silage. Ethanol content was greater in BG than in SC regardless of silage type. Aerobic deterioration occurred promptly in ensiling materials (nonensiled by-products and total mixed ration mixtures) and in silages stored alone; however, SCT and BGT silages resisted deterioration and no heating was found for more than 5.5 d regardless of storage period. Silages were stable even with high yeast populations at silo opening, whereas prolonged ensiling decreased yeast counts in the 2 total mixed ration silages. The denaturing gradient gel electrophoresis profiles appeared similar between SCT and BGT silages but not between SC and BG silages. Weissella spp. and Lactobacillus brevis were common in aerobically stable SCT and BGT silages, and Lactobacillus buchneri was detected only in BGT silage. Both L. brevis and L. buchneri were found in silage but not in ensiling materials. Several other lactic acid bacteria were also identified in SCT and BGT silages, but did not appear to be related to fermentation and aerobic stability.

DOI： 10.3168/jds.2007-0821

Variations in bacterial communities in laboratory-scale and big bale silos assessed by fermentation products, colony counts and denaturing gradient gel electrophoresis profiles Reviewed

Nishino N, Tanabe Y

Letters in Applied Microbiology 46 ( 3 ) 283 - 288 2008.3

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：BLACKWELL PUBLISHING

Aims: To assess the variation in bacterial communities in laboratory-scale and big bale silos.
Methods and Results: Wilted Italian ryegrass (628 g dry matter kg(-1)) was ensiled in vacuum-packed plastic pouches and big bales. Silos were opened after 3 months, and the fermentation products, colony counts and denaturing gradient gel electrophoresis (DGGE) profiles were determined. Eight samples were collected separately from a big bale, while one representative sample was taken from a plastic pouch. Significant variation was found between big bales in dry matter, ethanol, lactic acid, acetic acid and ammonia-N contents. No differences were shown between plastic pouches and big bales, except that more ethanol was produced in the former air-tight silos. Plastic pouches could resemble a specific silo and outer sampling sites of big bales based on fermentation products and DGGE profiles respectively.
Conclusions: Considerable variation in fermentation products may exist between big bale silos. Plastic pouches can serve as a model of big bale silos, although they do not provide information on the heterogeneity within and between bales.
Significance and Impact of the Study: Assessment of bacterial communities associated with ensiling can differ according to the criteria of fermentation products, colony counts and DGGE profiles.

DOI： 10.1111/j.1472-765X.2007.02306.x

Resistance to aerobic deterioration of total mixed ration silage: effect of ration formulation, air infiltration and storage period on fermentation characteristics and aerobic stability Reviewed

Fujin Wang, Naoki Nishino

Journal of the Science of Food and Agriculture 88 ( 1 ) 133 - 140 2008.1

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-BLACKWELL

BACKGROUND: High aerobic stability can be expected when wet brewers' grains are stored as a total mixed ration (TMR) silage. To understand the factors affecting the stability, the effects of ration formulation, air infiltration and storage period were studied.
RESULTS: A TMR containing wet brewers' grains, hay, maize, wheat bran, beet pulp and molasses was ensiled in laboratory silos for 14 and 56 days. The effects of hay species (lucerne or sudangrass) and air infiltration (Exp. 1) and of excluding one, two or three items from the six ingredients (Exp. 2) were examined. Ethanol was the main fermentation product in all TMR silages in this study. Aerobic deterioration occurred in 14 day silages prepared with sudangrass hay along with air infiltration (Exp. 1), and with the simplest recipe where three items (hay, maize and wheat bran) were excluded (Exp. 2). No deterioration occurred in 56 day silages regardless of ration formulation and air infiltration. Yeasts receded in 56 day silages, except with the simplest recipe, to the 102 cfu g(-1) level and remained undetectable in the presence of air (Exp. 2).
CONCLUSION: TMR silage can resist aerobic deterioration provided that a sufficient ensiling period has elapsed. Silages stored for only a few weeks may be susceptible to deterioration when air is infiltrated or where fewer ingredients are used in the TMR mixture. (C) 2007 Society of Chemical Industry.

DOI： 10.1002/jsfa.3057

Effect of aerobic exposure after silo opening on feed intake and digestibility of total mixed ration silage containing wet brewers grains or soybean curd residue Reviewed

WANG Fujin, NISHINO Naoki

Grassland science 54 ( 3 ) 164 - 166 2008

Resistance to aerobic deterioration of total mixed ration silage inoculated with and without homofermentative or heterofermentative lactic acid bacteria Reviewed

Naoki Nishino, Hirotaka Hattori

Journal of the Science of Food and Agriculture 87 ( 13 ) 2420 - 2426 2007.10

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS LTD

Wet brewers grains were stored as a total mixed ration (TMR) in laboratory silos with lucerne hay, cracked maize, sugar beet pulp, soya bean meal and molasses at 5:1:1:1:1:1 on fresh weight basis. The TMR mixture was inoculated with or without Lactobacillus casei or Lactobacillus buchneri to obtain silages with differing fermentation and stability after exposure to air. In the first experiment, ensiling was stopped at 10, 20 and 60 days, and the stability was tested for the following 7 days. Ethanol and lactic acid were the main products in untreated TMR silage, while addition of L. casei and L. buchneri increased lactic and acetic acid, respectively. No silages deteriorated in the presence of air over 7 days, regardless of inoculation, ensiling period and the level of yeasts determined at unloading. In the second experiment, silos were opened at 14 days and then subjected to aerobic stability test for 14 days. Resistance to deterioration was sustained in the untreated control, even with a high population (> 10(4) cfu g(-1)) of yeasts throughout the 14-day test. Spoilage was found in L. casei-treated silage at about 5 days, while increase of yeasts preceded the distinct heating (degradation). In L. buchneri-treated silage, no yeasts were detected at unloading or after exposure to air. These results suggest that substantial stability can be expected in TMR silage with or without inoculation of lactic acid bacteria. This property is not associated with the counts of yeasts at loading and the characteristics of silage such as alcoholic and lactic acid fermentation. (C) 2007 Society of Chemical Industry.

DOI： 10.1002/jsfa.2911

Biogenic amine production in grass, maize and total mixed ration silages inoculated with Lactobacillus casei or Lactobacillus buchneri Reviewed

N. Nishino, H. Hattori, H. Wada, E. Touno

Journal of Applied Microbiology 103 ( 2 ) 325 - 332 2007.8

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：BLACKWELL PUBLISHING

Aims: To investigate the effects of inoculating Lactobacillus casei or Lacobacillus buchneri on the production of biogenic amines (BA) in silage.
Methods and Results: Wilted festulolium (Lolium perenne x Festuca pratensis), whole crop maize or a total mixed ration, consisting of wet brewer grains, lucerne hay, cracked maize, sugarbeet pulp, soyabean meal and molasses, was ensiled with or without the inoculation of either L. casei (> 10(6) CFU g(-1)) or L. buchneri (> 10(6) CFU g(-1)). Silages were opened after 60 days of storage, and the concentrations of histamine, tyramine, putrescine and cadaverine were determined. The inoculation of L. casei decreased all the BA regardless of the silage type. The effects of L. buchneri varied between the three silages; the tyramine and putrescine were increased in maize but were lowered in festulolium. Histamine was reduced in fiestulolium and the by-products, whereas no change was found in the maize silage. None of the inoculant strains produced the four BA in a synthetic medium, accounting for the actual ensiling except for tyramine and putrescine in maize.
Conclusions: Wide variation would be found in the production of BA owing to the ensiling materials. The inoculation of L. casei can lower the BA concentration, while the effects of L. buchneri may vary considerably. The screening of BA-producing activity may help to reduce the risk of BA contamination in inoculated silage.
Significance and Impact of the Study: Strains of decarboxylase-negative L. buchneri can enhance the aerobic stability of silage without a concern regarding the large production of putrefactive BA.

DOI： 10.1111/j.1365-2672.2006.03244.x

Changes during ensilage in fermentation products, tea catechins, antioxidative activity and in vitro gas production of green tea waste stored with or without dried beet pulp Reviewed

Naoki Nishino, Tetsuya Kawai, Makoto Kondo

Journal of the Science of Food and Agriculture 87 ( 9 ) 1639 - 1644 2007.7

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS LTD

Wet green tea waste (GTW) was ensiled in laboratory silos with or without dried beet pulp at a ratio of 2:1 on a fresh weight basis. The silos were opened at 15, 30 and 60 days after anaerobic storage; fermentation products, tea catechins, antioxidative activity and in vitro gas production were determined. Acetic acid prevailed over the fermentation and the pH declined to below 4.2 when wet GTW was ensiled alone. Addition of dried beet pulp greatly enhanced lactic acid production and the pH further decreased to below 3.8 with no butyric acid being detected. Approximately 90% and 80% of (-)-epicatechin gallate and (-)-epigallocatechin gallate, respectively, were degraded in silage prepared from wet GTW alone, while the degradation was ameliorated in mixed silage to about 55% and 70%, respectively. Antioxidative activity was not markedly altered by ensiling and 70-80% of the initial activity was detected after 60 days. Changes were small in (-) -epigallocatechin, caffeine and total phenols with or without mixing with dried beet pulp. In vitro gas production was similar between materials and their silages, indicating that reduction of tea catechins would not elicit an improvement in digestibility. It is concluded that wet GTW can be ensiled successfully without bacterial inoculants when mixed with other materials containing certain sugars. Ensiling may degrade part of the components of tea catechins, while not affecting total phenols, antioxidative activity and digestibility of wet GTW. (c) 2007 Society of Chemical Industry.

DOI： 10.1002/jsfa.2842

Alcoholic fermentation and its prevention by Lactobacillus buchneri in whole crop rice silage Reviewed

N. Nishino, H. Hattori, Y. Kishida

Letters in Applied Microbiology 44 ( 5 ) 538 - 543 2007.5

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：BLACKWELL PUBLISHING

To evaluate the fermentation characteristics and the effects of Lactobacillus buchneri inoculation in ensiling whole crop rice.
Laboratory-scale silages were prepared from whole crop rice harvested at yellow-ripe stage. The crop was ensiled for 2 months with and without inoculation of L. buchneri at 10(4), 10(5) and 10(6) CFU g(-1). The effect of prolonged ensiling was also studied by using the same crop; the silos were opened at 1, 3, 6 and 12 months, while the inoculation was made at 10(5) CFU g(-1). Enhanced alcoholic fermentation was found in untreated silage; the sum of ethanol and 2,3-butanediol were seven times higher at 2 months than those of lactic and volatile fatty acids, while the differences were diminished at 12 months owing to the reduction of ethanol in the late ensiling period. Inoculation of L. buchneri inhibited the alcohols; however, ethanol yet prevailed over the fermentation until 6 months, after which acetic acid became the main product in the inoculated silage. Regardless of inoculation and ensiling period, yeasts were not found in whole crop rice silage.
Substantial amounts of ethanol and 2,3-butanediol would be produced in silage prepared from whole crop rice. The alcoholic fermentation can be suppressed when inoculated with L. buchneri.
Inoculation of L. buchneri could be an option to prevent ethanol fermentation in silage.

DOI： 10.1111/j.1472-765X.2006.02105.x

Ethanol and 2,3-butanediol production in whole-crop rice silage. Reviewed

Nishino, N, Shinde, S

Grassland Science 53 ( 3 ) 196 - 198 2007

Potential nutritive value of selected browse species from Kenya using in vitro gas production technique and polyethylene glycol. Reviewed

Osuga, I. M, Abdulrazak, S. A, Nishino, N, Ichinohe, T, Fujihara, T

Livestock Research for Rural Development 2006

Ensiling characteristics and aerobic stability of direct-cut and wilted grass silages inoculated with Lactobacillus casei or Lactobacillus buchneri Reviewed

N Nishino, E Touno

Journal of the Science of Food and Agriculture 85 ( 11 ) 1882 - 1888 2005.8

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-BLACKWELL

Two experiments were conducted to study the effects of wilting and inoculating Lactobacillus casei or Lactobacillus buchneri on the fermentation and aerobic stability of grass silages. Chopped Italian ryegrass (IR) and Festulolium (FE) were ensiled with or without wilting and added L casei (> 10(6) Cfu g(-1) fresh matter (FM)) or L buchneri (> 10(6) cfu g(-1) FM). Silos were opened after 120 days and microbial counts, fermentation products and aerobic stability were determined. Addition of L casei increased lactic acid and decreased acetic acid and dry matter loss regardless of wilting and forage species. Inoculation of L buchneri decreased lactic acid and increased acetic acid and 1,2-propanediol, while the effects appeared greater in direct-cut than in wilted grass silages. Although 1,2-propanediol accumulated in FE silage, the diol was degraded further to propionic acid and 1-propanol in IR silage. The activity of 1,2-propanediol degradation was lowered when IR was wilted prior to ensiling; 1,2-propanediol remained and the production of propionic acid and 1-propanol was less than one-third of that in direct-cut silage. Regardless of forage species, addition of L buchneri increased dry matter loss compared with the untreated control, whereas the loss was not significant in. wilted silages. Ammonia production was increased by L buchneri in direct-cut but not in wilted silages. No heating was observed with or without inoculation in direct-cut IR silage after exposure to air. Other silages were deteriorated when L buchneri was not inoculated, while the spoilage was accelerated when L casei was added to FE. (c) 2005 Society of Chemical Industry.

DOI： 10.1002/jsfa.2189

Microbial counts, fermentation products, and aerobic stability of whole crop corn and a total mixed ration ensiled with and without inoculation of Lactobacillus casei or Lactobacillus buchneri Reviewed

N Nishino, H Wada, M Yoshida, H Shiota

Journal of Dairy Science 87 ( 8 ) 2563 - 2570 2004.8

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：AMER DAIRY SCIENCE ASSOC

Whole crop corn (DM 29.2%) and a total mixed ration (TMR, DM 56.8%) containing wet brewers grains, alfalfa hay, dried beet pulp, cracked corn, soybean meal, and molasses at a ratio of 5: 1: 1: 1: 1: 1 on fresh weight basis, were ensiled with and without Lactobacillus casei or Lactobacillus buchneri in laboratory silos. The effects of inoculation on microbial counts, fermentation products, and aerobic stability were determined after 10 and 60 d. Untreated corn silage was well preserved with high lactic acid content, whereas large numbers of remaining yeasts resulted in low stability on exposure to air. Inoculation with L. casei suppressed heterolactic fermentation, but no improvements were found in aerobic stability. The addition of L. buchneri markedly enhanced the aerobic stability, while not affecting the DM loss andNH(3)-N production. Large amounts of ethanol were found when the TMR was ensiled, and the content of ethanol overwhelmed that of lactic acid in untreated silage. This fermentation was related to high yeast populations and accounted for a large loss of DM found in the initial 10 d. The ethanol production decreased when inoculated with L. casei and L. buchneri, but the effects diminished at 60 d of ensiling. Inoculation with L. buchneri lowered the yeasts in TMR silage from the beginning of storage; however, the populations decreased to undetectable levels when stored for 60 d, regardless of inoculation. No heating was observed in TMR silage during aerobic deterioration test for 7 d. This stability was achieved even when a high population of yeasts remained and was not affected by either inoculation or ensiling period. The results indicate that inoculation with L. buchneri can inhibit yeast growth and improve aerobic stability of corn and TMR silage; however, high stability of TMR silage can be obtained even when no treatments were made and high population (> 10(5) cfu/g) of yeasts were detected.

DOI： 10.3168/jds.S0022-0302(04)73381-0

Enhanced lactic acid fermentation of silage by the addition of green tea waste Reviewed

M Kondo, N Naoki, K Kazumi, H Yokota

Journal of the Science of Food and Agriculture 84 ( 7 ) 728 - 734 2004.5

Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS LTD

The effects of green tea waste (GTW) addition on the ensiling of forage were investigated. Wet and dried GTW added at 10, 50, 100 and 200g kg(-1) of fresh matter (FM) and at 2, 10 and 20g kg(-1) FM, respectively, decreased pH and increased lactic acid concentration of the silages, whereas the butyric acid concentration and ammonia nitrogen content, as a proportion of a total nitrogen, were lowered, compared with silage without additives (control). To investigate the effect of GTW-associated LAB on silage fermentation, wet GTW was sterilized by autoclaving or gamma irradiation and added at 50 g kg(-1) FM. The silages made with sterilized GTW showed higher lactic acid concentrations, and lower pH and butyric acid concentrations than controls. The counts of lactic acid bacteria (LAB) were higher in silages made with sterilized GTW than control until 10 days after ensiling. The enhanced lactic acid fermentation was not found when green tea polyphenols (GTP) were added. These data suggested that GTW could enhance LAB growth and lactic acid production of silage, particularly when added at 50 g kg(-1) FM in a wet form or at the equivalent in a dry form. Although neither GTW-associated LAB nor GTP accounted for the enhancement of lactic acid fermentation, GTW would possibly supply some nutrients which are heat-stable and effective for LAB growth during silage fermentation.. (C) 2004 Society of Chemical Industry.

DOI： 10.1002/jsfa.1726

Evaluation of fermentation and aerobic stability of wet brewers' grains ensiled alone or in combination with various feeds as a total mixed ration Reviewed

N Nishino, H Harada, E Sakaguchi

Journal of the Science of Food and Agriculture 83 ( 6 ) 557 - 563 2003.5

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS LTD

Wet brewer's grains (BG; 200 g kg(-1) dry matter(DM)) were ensiled alone or in combination with various dry feeds as a total mixed ration (TMR; 540 g kg(-1) DM) in laboratory silos. Ensilage was stopped at 40 days (experiment 1) or at 5, 20, 40 and 60 days (experiment 2) after storage. The composition of soluble sugars and the profiles of fermentation products were determined. The 60 day silages were subjected to aerobic deterioration and changes in silage temperature were recorded for 7 days. A rapid pH fall coupled with accelerated lactic acid production was observed in BG silage; soluble sugars, mainly composed of maltose and raffinose, completely disappeared within 5 days of ensiling. Prolonged storage, however, decreased lactic acid concentration and increased acetic acid greatly. Appreciable amounts of propionic acid and I-propanol were also produced in BG silage in the late stages of fermentation. When TMR silage was made, the decline in pH and the increase in lactic acid were delayed due to the low moisture content. Acetic acid production was enhanced from the beginning of storage, and mannitol accumulated in TMR silage. Yeast numbers were lower in TMR than in BG silage, but the changes were limited in the late stages of fermentation. When exposed to air, the temperature of BG silage increased after 3 days, whereas that of TMR silage appeared stable during the 7 days test. The results indicate that BG is a suitable by-product for ensiling and, when ensiled with various feeds as a TMR, improved stability against aerobic deterioration can be expected. (C) 2003 Society of Chemical Industry.

DOI： 10.1002/jsfa.1395

Accumulation of 1,2-propanediol and enhancement of aerobic stability in whole crop maize silage inoculated with Lactobacillus buchneri Reviewed

N Nishino, M Yoshida, H Shiota, E Sakaguchi

Journal of Applied Microbiology 94 ( 5 ) 800 - 807 2003

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：BLACKWELL PUBLISHING LTD

Aims: To assess the effects of inoculation of Lactobacillus buchneri on the ensiling properties and aerobic stability of maize silage.
Methods and Results: Chopped whole crop maize was ensiled in 0.5 litre airtight polyethylene bottles (0.4 kg per bottle) and in double-layered, thin polyethylene bags (15 kg per bag), with or without inoculation of Lact. buchneri . The silos were stored for two to four months and the chemical composition, microbial numbers and aerobic stability were determined. Inoculation lowered lactic acid and yeasts, and increased acetic acid and pH value, resulting in improved aerobic stability of the silages. Inoculated silages produced 1,2-propanediol, the content of which increased as ensiling was prolonged, and nearly 50 g kg(-1) dry matter had accumulated after four months of storage. The effects of inoculation, however, were much less pronounced in silages prepared in bags. Mannitol was found in all silages; the production was lowered by Lact. buchneri treatment and appeared to be unrelated to the accumulation of 1,2-propanediol.
Conclusions: Inoculation of Lact. buchneri occasionally causes accumulation of 1,2-propanediol in silages without further degradation into propionic acid and 1-propanol.
Significance and Impact of the Study: Substantial amounts of 1,2-propanediol could be consumed by ruminants when fed on silages inoculated with Lact. buchneri . In addition to increasing acetic acid, attention needs to be paid to 1,2-propanediol because the two fermentation products might affect the intake and utilization of silage-based diets.

DOI： 10.1046/j.1365-2672.2003.01810.x

Content of phenolics and tannins in leaves and pods of some Acacia and Dichrostachys species and effects on in vitro digestibility Reviewed

CDK Rubanza, M Shem, R Otsyina, N Nishino, T Ichinohe, T Fujihara

Journal of Animal and Feed Sciences 12 ( 3 ) 645 - 663 2003

Language：English Publishing type：Research paper (scientific journal) Publisher：KIELANOWSKI INST ANIMAL PHYSIOLOGY NUTRITION

Four browse legume foliages (leaves and pod fruits) from three species of Acacia (A. polyacantha, A. tortilis, A. nilotica) and Dichrostachys sp. native to Tanzania were evaluated for nutritive potential to establish levels and effect of normal phenolics and tannins antinutritive factors (ANFs) on in vitro feed digestibility (IVD). Total extractable phenolics (TP), tannins (TT) were estimated by Folin-Ciocalteu assay, and condensed tannins (CT) by butanol/HCl assay. In leaves, total proanthocyanidins were assayed into constituent anthocyanidins' flavonoids; by high performance liquid chromatography (HPLC). Organic matter degradability (OMD) was estimated in vitro by gas production technique. Adverse effects of tannin's ANF on IVD were assessed by polyethylene glycol (PEG) tannin bioassay. Crude protein (CP) varied (P<0.05) between fodder species and foliages. Leaves had (P<0.05) higher CP (141-194 g/kg DM) in Dichrostachys sp. and A polyacantha, respectively, compared to pods (133-142 g/kg DM) in A. tortilis and Dichrostachys sp., respectively. Neutral detergent fibre (NDF), acid detergent fibre (ADF) and acid detergent lignin (ADL) varied (P<0.05) between fodder species and foliages. A. nilotica had lowest (P<0.05) NDF, ADF and ADL (182, 68 and 44 g/kg DM respectively). A. polyacantha and Dichrostachys sp. had highest (P<0.05) NDF and ADF (416 and 146 g/kg DM, respectively). Pods had higher (P<0.05) fibre values than leaves. Leaves had higher (P<0.05) TP, TT, and CT (162 138, and 55 mg/g DM), than pods (112, 105 and 35 mg/g DM), respectively. Proanthocyanidin flavonoids (mg/g DM) varied (P<0.05) from 0.062 (A. nilotica) to 5.288 (A. tortilis), 0.188 (A. tortilis) to 4.179 (A. polyacantha), and 0.009 (A. nilotica) to 4.392 (Dichrostachys sp.) for delphinidins, cyanidins and pelargonidin, respectively. The browse foliages had relatively high tannin content greater than 5% DM, a beneficial level in animal feeding and nutrition, and thus could impair feed digestibility. OMD varied significantly (P<0.05) both between fodder species and foliage components. Pods had higher (P<0.05) gas production (GP) and OMD than leaves. Addition of PEG improved (P<0.05) gas production, IVD and metabolizable energy (ME) in leaves and pods by binding tannins. Leaves had higher response on gas production and IVD due to PEG treatment compared to pods except A. nilotica. Improved gas production and digestibility due to PEG treatment indicate nutritive potential in browse fodder previously depressed by tannin ANFs.
Therefore, phenolics and tannins ANFs could limit utilization of browse fodder nutritive potential as supplements to ruminants consuming low quality roughages. Utilization of browse legume fodder could be optimized through reduction of tannin antinutritive activity. Under farmers' conditions, optimal utilization of browse could be achieved through feeding a mixture of tanniferous browse with other feeds especially high in nitrogen to dilute tannin antinutritive activity.

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