Faculty Profiles - NISHINO Naoki

Papers - NISHINO Naoki

Division display >> ／ All the affair displays 1 - 83 of about 83

Cyclic oligosaccharide-induced modulation of immunoglobulin A reactivity to gut bacteria contributes to alterations in the bacterial community structure

Miyamoto, T., Tsuruta, T., Teraoka, M., Wang, T., Nishino, N.

Nutrients 16 2824 - 2824 2024.8

Publishing type：Research paper (scientific journal) Publisher：MDPI AG

Immunoglobulin A (IgA) is a major gut antibody that coats commensal gut bacteria and contributes to shaping a stable gut bacterial composition. Although previous studies have shown that cyclic oligosaccharides, including cyclic nigerosyl-1,6-nigerose (CNN) and cyclodextrins (CDs, including αCD, βCD, and γCD), alter the gut bacterial composition, it remains unclear whether cyclic oligosaccharides modify the IgA coating of gut bacteria, which relates to cyclic oligosaccharide-induced alteration of the gut bacterial composition. To address this issue, mice were maintained for 12 weeks on diets containing CNN, αCD, βCD, or γCD; the animals’ feces were evaluated for their bacterial composition and the IgA coating index (ICI), a measure of the degree of IgA coating of bacteria. We observed that the intake of each cyclic oligosaccharide altered the gut bacterial composition, with changes in the ICI found at both the phylum and genus levels. The ICI for Bacillota, Lachnospiraceae NK4A136 group, UC Lachnospiraceae, and Tuzzerella were significantly and positively correlated with the relative abundance (RA) in total bacteria for these bacteria; in contrast, significant correlations were not seen for other phyla and genera. Our observations suggest that cyclic oligosaccharide-induced modulation of the IgA coating of gut bacteria may partly relate to changes in the community structure of the gut bacteria.

DOI： 10.3390/nu16172824

Unabsorbed fecal fat content correlates with a reduction of immunoglobulin A coating of gut bacteria in high-lard diet-fed mice. International journal

Katsumata, E., Tsuruta, T., Sonoyama, K., Yoshida, T., Sasaki, M., Teraoka, M., Wang, T., Nishino, N.

Molecular Nutrition & Food Research 68 e2400078 2024.8

Language：English Publishing type：Research paper (scientific journal)

SCOPE: Immunoglobulin A (IgA) selectively coats gut bacteria and contributes to regulatory functions in gastrointestinal inflammation and glucose metabolism. Excess intake of lard leads to decrease in the IgA coating of gut bacteria, although the underlying mechanisms remain unknown. This study validates how unabsorbed fat derived from a high-lard diet in the gut affects the IgA coating of bacteria, as assessed in mouse models using three types of dietary fat (lard, medium-, and long-chain triglycerides [MLCTs], and medium-chain triglycerides [MCTs]) exhibiting different digestibilities. METHODS AND RESULTS: C57BL/6J mice are maintained on diets containing lard, MLCTs, or MCTs at 7% or 30% w/w for 10 weeks (n = 6 per group). The fecal fatty acid concentration is measured to quantify unabsorbed fat content. The ratio of IgA-coated bacteria to total bacteria (IgA coating ratio) in the feces is measured by flow cytometry. Compared to lard-fed mice, MLCT- and MCT-fed mice exhibit lower fecal concentrations of palmitic acid, stearic acid, and oleic acid and higher IgA coating ratios at both 7% and 30% dietary fat, and these parameters exhibit significant negative correlations. CONCLUSION: Unabsorbed fat content in the gut may result in attenuated IgA coating of bacteria in high-lard diet-fed mice.

DOI： 10.1002/mnfr.202400078

Variations in milk, udder skin, and fecal microbiota and their relationships with blood metabolites and milk composition in dairy cows. Reviewed

AoDaohu, Akada, K., Gathinji, P.K., Nishino, N.

Letters in Applied Microbiology 77 ovae014 2024.3

Commercial concentrate supplementation in Phan Rang sheep diets: Effects on digestibility traits, growth and carcass performance. Reviewed

Nguyen, H.V., Nguyen, T.M., Dinh, V.D., Van, N.P., Tran, N.L., Le, T.H., Le, D.T., Vo, T.M., Ngo, M.D., Bui, V.L., Nguyen, X.B., Ton, N.M.T., Nishino, N.

Advances of Animal and Veterinary Sciences 12 1 - 8 2024.1

In vitro assessment of the relationships between the digestion of different types of rice straw and bacterial community in the rumen. Reviewed

68 372 - 382 2023.9

Cyclic nigerosylnigerose attenuates high-fat diet induced fa;dposition;colonic inflammation;d;abnormal;glucose metabolism;modifies gut immunoglobulin A reactivity to commensal bacteria. Reviewed International journal

Tsuruta, T, Sonoyama, K, Miyamoto, T, Nguyen, Q.D, Mizote, A, Sasou, H, Shiotsu, M, Teraoka, M, Nishino, N

Molecular Nutrition and Food Research 67 202200389 2023.7

Language：English Publishing type：Research paper (scientific journal)

SCOPE: High-fat diet (HFD) intake induces gut dysbiosis, inflammation in the peripheral tissues, and a reduction in immunoglobulin A (IgA) coating of gut bacteria, which is related to HFD-induced insulin resistance (IR). This study evaluates the effect of cyclic nigerosylnigerose (CNN), a dietary fiber that prevents gut inflammation and promotes IgA coating of gut bacteria, on the above-mentioned HFD-induced disorders. METHODS AND RESULTS: Balb/c mice are fed an HFD and administered CNN for 20 weeks. CNN administration reduces mesenteric adipose tissue weight, colonic tumor necrosis factor α (TNFα) mRNA expression, and serum endotoxin levels and ameliorates HFD-induced abnormal glucose metabolism. Additionally, CNN administration promotes gut bacteria-specific IgA secretion and alters IgA reactivity to gut bacteria. The alterations of IgA reactivity to specific bacteria such as Erysipelatoclostridium, Escherichia, Faecalibaculum, Lachnospiraceae genera, and Stenotrophomonas are correlated with mesenteric adipose tissue weight, colonic TNFα mRNA expression, serum endotoxin levels, and a homeostasis model assessment for IR. CONCLUSION: CNN-induced alterations in IgA reactivity to gut bacteria may be related to the suppression of HFD-induced fat deposition, colonic inflammation, endotoxemia, and IR. These observations indicate that dietary fiber that modulates IgA reactivity to gut bacteria may be useful in preventing HFD-induced disorders.

DOI： 10.1002/mnfr.202200389

Modulatory effects of A1 milk, A2 milk, soy, and egg proteins on gut microbiota and fermentation. Reviewed

Nuomin, Baek, R, Tsuruta, T, Nishino, N

Microorganisms 11 1194 2023.5

Monitoring the milk composition, milk microbiota, and blood metabolites of Jersey cows throughout a lactation period. Reviewed

Gathinji, P.K., Yousofi, Z., Akada, K., Wali, A., Nishino, N.

Veterinary Sciences 10 226 2023.3

Frequency of β-casein gene polymorphisms in Jersey cows in western Japan. Reviewed

Nuomin, Nguyen, D.Q, Aodaohu, Nishino, N

Animals 12 2076 2022.8

Bacterial and fungal microbiota of total mixed ration silage stored at various temperatures. Reviewed International journal

Wali, A, Hou, J, Tsuruta, T, Nishino, N

Journal of Applied Microbiology 133 ( 2 ) 579 - 590 2022.8

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)

AIMS: To obtain insights into how bacterial and fungal microbiota and fermentation products composition are affected by storage temperature for TMR silage, which can be manufactured year-round. METHODS AND RESULTS: TMR silage was stored at 10°C, 25°C, ambient temperature (AT; 20-35°C) and 40°C. Lactic acid production was delayed when stored at 10°C, and acid production stagnated after 2 weeks when stored at 40°C. The patterns of acetic acid and ethanol production were inversely related, with ethanol production promoted at 10°C and 25°C and acetic acid production promoted at AT and 40°C. The bacterial diversity was reduced in TMR silage with high lactic acid and acetic acid content, and the fungal diversity was reduced in TMR silage with high ethanol content. CONCLUSIONS: The intensity of lactic acid production was accounted for by the high abundance of Lactobacillus, and its stagnated production at a substantially high storage temperature was related to an increased abundance of Bacillus. The enhanced production of acetic acid or ethanol can be explained by differences in the fungal microbiota. SIGNIFICANCE AND IMPACT OF THE STUDY: The integrated analysis of bacterial and fungal microbiota can provide in-depth insights into the impact of storage temperature on TMR silage fermentation.

DOI： 10.1111/jam.15582

Fecal, milk, uterine, airborne dust, and water microbiota in dairy farms in southern Vietnam: A pilot study. Reviewed

Tran, T.M.T, Tran, H.D, Nguyen, T.T, Acosta, T.J, Tsuruta, T, Nishino, N, Duong, T.H

Advances of Animal and Veterinary Sciences 10 ( 7 ) 1525 - 1531 2022.7

Publishing type：Research paper (scientific journal)

Six sets of fecal, milk, and uterine samples from dairy cows and two sets of airborne dust and water samples from the cowshed were collected from two farms in Ho Chi Minh City at 20–40 days postpartum. The microbiota was characterized using Illumina sequencing of the V4 hypervariable region of the 16S rRNA genes. The predominant species in the fecal microbiota was not as predominant in the milk microbiota, but occasionally appeared in the uterine microbiota. The microbiota from the airborne dust and water samples did not show a relation with either the milk or the uterine microbiota, whereas Moraxellaceae, the most abundant family in the airborne dust and water microbiota, was detected at low proportions in the milk microbiota. The farm-to-farm difference was more apparent for the milk microbiota than for the fecal and uterine microbiota. It is noteworthy that Dermacoccaceae, Bacillaceae, and Methylobacteriaceae were the prevalent families in the microbiota of several milk and uterine samples; however, these findings did not help understand the role of the microbiota in fertility and subsequent conception. Despite the small number of dairy cows examined in this study, this report is the first to characterize the fecal, milk, uterine, airborne dust, and water microbiota in dairy farms in southern Vietnam.

DOI： 10.17582/journal.aavs/2022/10.7.1525.1531

Bacterial and fungal microbiota of guinea grass silage shows various levels of acetic acid fermentation Reviewed

Hou, J, Nishino, N

Fermentation 8 10 2022.1

Aicda deficiency exacerbates high-fat diet-induced hyperinsulinemia but not gut dysbiosis in mice Reviewed

Tsuruta, T, Muhomah, T.A, Sonoyama, K, Nguyen, Q.D, Takase, Y, Nishijima, A, Himoto, S, Katsumata, E, Nishino, N

Nutrition Research 93 15 - 26 2021.9

Cecum microbiota in rats fed soy, milk, meat, fish, and egg proteins with prebiotic oligosaccharides Reviewed International journal

Sivixay, S, Bai, G, Tsuruta, T, Nishino, N

AIMS Microbiology 7 ( 1 ) 1 - 12 2021.1

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)

Diet is considered the most influential factor in modulating the gut microbiota but how dietary protein sources differ in their modulatory effects is not well understood. In this study, soy, meat (mixture of beef and pork), and fish proteins (experiment 1) and soy, milk (casein), and egg proteins (experiment 2) were fed to rats with cellulose (CEL) and raffinose (RAF); the microbiota composition and short-chain fatty acid concentration in the cecum were determined. Egg protein feeding decreased the concentration of acetic acid and the richness and diversity of the cecum microbiota. RAF feeding increased the concentrations of acetic and propionic acids and decreased the richness and diversity of the cecum microbiota. When fed with CEL, the abundance of Ruminococcaceae and Christensenellaceae, Akkermansiaceae and Tannerellaceae, and Erysipelotrichaceae enhanced with soy protein, meat and fish proteins, and egg protein, respectively. The effects of dietary proteins diminished with RAF feeding and the abundance of Bifidobacteriaceae, Erysipelotrichaceae, and Lachnospiraceae increased and that of Ruminococcaceae and Christensenellaceae decreased regardless of the protein source. These results indicate that, although the effect of prebiotics is more robust and distinctive, dietary protein sources may influence the composition and metabolic activities of the gut microbiota. The stimulatory effects of soy, meat, and egg proteins on Christensenellaceae, Akkermansiaceae, and Erysipelotrichaceae deserve further examination to better elucidate the dietary manipulation of the gut microbiota.

DOI： 10.3934/microbiol.2021001

An investigation of seasonal variations in the microbiota of milk, feces, bedding, and airborne dust Reviewed

Nguyen, T.T, Wu, H, Nishino, N

Asian-Australasian Journal of Animal Sciences 33 ( 11 ) 1858 - 1865 2020.11

Bacterial and fungal microbiota associated with the ensiling of wet soybean curd residue under prompt and delayed sealing conditions Reviewed

Wali, A, Nishino, N

Microorganisms 8 9 2020.9

Cyclic nigerosylnigerose ameliorates DSS-induced colitis with restoration of goblet cell number and increase in IgA reactivity against gut microbiota in mice Reviewed

Tsuruta, T, Katsumata, E, Mizote, A, Hou, J, Muhomah, T.A, Nishino, N

Bioscience of Microbiota, Food and Health 39 ( 3 ) 188 - 196 2020.7

Language：English Publishing type：Research paper (scientific journal)

Cyclic nigerosylnigerose (CNN) is a cyclic oligosaccharide. Oral administration of CNN promotes immunoglobulin A (IgA) secretion in the gut. IgA is a major antibody secreted into the gut and plays a crucial role in suppressing gut inflammation due to commensal gut microbiota. To investigate the effect of administration of CNN to promote IgA secretion on gut inflammation, experimental colitis was induced with dextran sulfate sodium (DSS) in Balb/c mice after 6 weeks of CNN pre-feeding. The severity of colitis was evaluated based on a disease activity index (DAI), the gene expression of inflammatory cytokines, and a histological examination. The CNN-treated mice with DSS-induced colitis (CNN-DSS group) showed significantly lower DAI scores and mRNA levels of interleukin-1 compared with the CNN-untreated mice with DSS-induced colitis (DSS group). Histological examination of the colon revealed that the pathological score was significantly lower in the CNN-DSS group compared with the DSS group due to the reduced infiltration of immune cells. The number of goblet cells was significantly higher in the CNN-DSS group compared with the DSS group. The IgA concentration and the ratio of microbiota coated with IgA were evaluated in the cecal content. Although there was no difference in the IgA concentration among groups, a higher proportion of cecal microbiota were coated with IgA in the CNN-DSS group compared with that in the DSS group. These results suggest that CNN might preserve goblet cells in the colon and promote IgA coating of gut microbiota, which synergistically ameliorate gut inflammation in mice with DSS-induced colitis.

DOI： 10.12938/bmfh.2020-012

Examination of milk microbiota, fecal microbiota, and blood metabolites of Jersey cows in cool and hot seasons Reviewed International journal

Nguyen, Q.D, Tsuruta, T, Nishino, N

Animal Science Journal 91 ( 1 ) e13441 2020.1

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal)

Microbiota of individual cow milk, bulk tank milk, and feces of Jersey cows were examined. Samples were collected from two farms (F1 and F2) in cool (November, Nov) and hot (July, Jul) seasons. Milk yield and milk composition were similar between the two farms and between the two seasons. Prevalent taxa of the fecal microbiota, i.e. Ruminococcaceae, Bacteroidaceae, Lachnospiraceae, Rikenellaceae, and Clostridiaceae, were unaffected by the farm and season. Relative abundance of milk microbiota for Pseudomonadaceae, Enterobacteriaceae, and Streptococcaceae (F1 > F2) and Lactobacillaceae, Bifidobacteriaceae, and Cellulomonadaceae (F1 < F2) were different between the two farms, and those for Staphylococcaceae, Bacillaceae, Ruminococcaceae, and Veillonellaceae (Nov < Jul) and Methylobacteriaceae and Moraxellaceae (Nov > Jul) were different between the two seasons. The microbiota of bulk tank milk was numerically different from that of individual cow milk. Principal coordinate analysis indicated that the milk microbiota was unrelated to the fecal microbiota. The finding that relative abundance of Pseudomonadaceae and Moraxellaceae appeared greater than those reported for Holstein milk suggested that higher protein and fat content may result in a greater abundance of proteolytic and lipolytic taxa in Jersey cow milk.

DOI： 10.1111/asj.13441

The relationship between uterine, fecal, bedding, and airborne dust microbiota from dairy cows and their environment: A pilot study Reviewed

Nguyen, T.T, Miyake, A, Tran, T.T.M, Tsuruta, T, Nishino, N

Animals 9 ( 12 ) 1007 2019.12

High-fat diet reduces the level of secretory immunoglobulin A coating of commensal gut microbiota Reviewed

Muhomah, T.A, Nishino, N, Katsumata, E, Wu, H, Tsuruta, T

Bioscience of Microbiota, Food and Health 38 ( 2 ) 55 - 64 2019.4

Rumen fluid, feces, milk, water, feed, airborne dust, and bedding microbiota in dairy farms managed by automatic milking systems Reviewed

Wu, H, Nguyen, Q.D, Tran, T.T.M, Tang, M.T, Tsuruta, T, Nishino, N

Animal Science Journal 90 ( 3 ) 445 - 452 2019.3

Dietary soy, meat, and fish proteins modulate the effects of prebiotic raffinose on composition and fermentation of gut microbiota in rats Reviewed

Gaowa Bai, Takeshi Tsuruta, Naoki Nishino

International Journal of Food Sciences and Nutrition 69 ( 4 ) 480 - 487 2018.5

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Taylor and Francis Ltd

Soy, meat (mixture of pork and beef), and fish proteins were fed to rats with and without prebiotic raffinose (RAF), and the composition and fermentation of gut microbiota were examined. Bifidobacterium spp. populations were higher, and propionic acid concentration was lower in soy protein-fed than meat protein-fed rats. Likewise, Enterobacteriaceae populations were higher in fish protein-fed rats than other rats. RAF feeding increased Bifidobacterium spp. and decreased Faecalibacterium prausnitzii populations regardless of the dietary protein source. Interactions between dietary proteins and RAF were shown for Lactobacillus spp. and Clostridium perfringens group
the increase of Lactobacillus spp. populations by RAF was seen only for soy protein-fed rats, whereas the reduction of C. perfringens group by RAF was evident in fish and meat protein-fed rats. It is concluded that dietary proteins may differentially modulate the effects of prebiotic oligosaccharides on gut fermentation and microbiota, with differences observed between plant and animal proteins.

DOI： 10.1080/09637486.2017.1382454

Effects of wilting and lactic acid bacteria inoculation on fermentation and microbial community of elephant grass silage produced in Vietnam Reviewed

Thi Minh Tu Tran, Mui Thi Nguyen, Huu Van Nguyen, Naoki Nishino

Grassland Science 64 ( 2 ) 151 - 155 2018.4

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Blackwell Publishing Inc.

Direct-cut (dry matter [DM] 165 g kg−1) and wilted (DM 250 g kg−1) elephant grass silages were prepared in Hue, Vietnam, with and without a mixture of Lactobacillus paracasei and Lactococcus lactis (LP+LC) or Lactobacillus buchneri (LB). The silages were stored for 4 months, and fermentation products and aerobic stability were determined. The microbial community was assessed using denaturing gradient gel electrophoresis and quantitative polymerase chain reaction. Acetate was predominantly produced during the fermentation of control and LB-inoculated silages, whereas the content was not sufficient to inhibit aerobic spoilage for 7 days in control silage. The LP+LC inoculation greatly enhanced lactate content, suppressed the alcohol content, and did not improve aerobic stability or decrease the total fungal population. The bacterial community and L. buchneri populations were similar between control and LB-inoculated silages
however, Kluyveromyces delphensis was undetectable and the total fungal population was lowered in LB-inoculated silage.

DOI： 10.1111/grs.12187

Identification of lactic acid bacteria in the feces of dairy cows fed whole crop maize silage to assess the survival of silage bacteria in the gut Reviewed

Hongyan Han, Chao Wang, Yanbing Li, Zhu Yu, Qingfang Xu, Guangpeng Li, Tang Thuy Minh, Naoki Nishino

Animal Science Journal 89 ( 1 ) 97 - 104 2018.1

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：Blackwell Publishing

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 106–7 and 103–4 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.

DOI： 10.1111/asj.12896

Variability, stability, and resilience of fecal microbiota in dairy cows fed whole crop corn silage Reviewed

Minh Thuy Tang, Hongyan Han, Zhu Yu, Takeshi Tsuruta, Naoki Nishino

Applied Microbiology and Biotechnology 101 ( 16 ) 6355 - 6364 2017.8

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER

The microbiota of whole crop corn silage and feces of silage-fed dairy cows were examined. A total of 18 dairy cow feces were collected from six farms in Japan and China, and high-throughput Illumina sequencing of the V4 hypervariable region of 16S rRNA genes was performed. Lactobacillaceae were dominant in all silages, followed by Acetobacteraceae, Bacillaceae, and Enterobacteriaceae. In feces, the predominant families were Ruminococcaceae, Bacteroidaceae, Clostridiaceae, Lachnospiraceae, Rikenellaceae, and Paraprevotellaceae. Therefore, Lactobacillaceae of corn silage appeared to be eliminated in the gastrointestinal tract. Although fecal microbiota composition was similar in most samples, relative abundances of several families, such as Ruminococcaceae, Christensenellaceae, Turicibacteraceae, and Succinivibrionaceae, varied between farms and countries. In addition to the geographical location, differences in feeding management between total mixed ration feeding and separate feeding appeared to be involved in the variations. Moreover, a cow-to-cow variation for concentrateassociated families was demonstrated at the same farm; two cows showed high abundance of Succinivibrionaceae and Prevotellaceae, whereas another had a high abundance of Porphyromonadaceae. There was a negative correlation between forage-associated Ruminococcaceae and concentrate-associated Succinivibrionaceae and Prevotellaceae in 18 feces samples. Succinivibrionaceae, Prevotellaceae, p-253418B5, and Spirochaetaceae were regarded as highly variable taxa in this study. These findings help to improve our understanding of variation and similarity of the fecal microbiota of dairy cows with regard to individuals, farms, and countries. Microbiota of naturally fermented corn silage had no influence on the fecal microbiota of dairy cows.

DOI： 10.1007/s00253-017-8348-8

Comparative microbiota assessment of wilted Italian ryegrass, whole crop corn, and wilted alfalfa silage using denaturing gradient gel electrophoresis and next-generation sequencing Reviewed

Kuikui Ni, Tang Thuy Minh, Tran Thi Minh Tu, Takeshi Tsuruta, Huili Pang, Naoki Nishino

Applied Microbiology and Biotechnology 101 ( 4 ) 1385 - 1394 2017.2

Authorship：Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER

The microbiota of pre-ensiled crop and silage were examined using denaturing gradient gel electrophoresis (DGGE) and next-generation sequencing (NGS). Wilted Italian ryegrass (IR), whole crop corn (WC), and wilted alfalfa (AL) silages stored for 2 months were examined. All silages contained lactic acid as a predominant fermentation product. Across the three crop species, DGGE detected 36 and 28 bands, and NGS identified 253 and 259 genera in the pre-ensiled crops and silages, respectively. The NGS demonstrated that, although lactic acid bacteria (LAB) became prevalent in all silages after 2 months of storage, the major groups were different between crops: Leuconostoc spp. and Pediococcus spp. for IR silage, Lactobacillus spp. for WC silage, and Enterococcus spp. for AL silage. The predominant silage LAB genera were also detected by DGGE, but the presence of diverse non-LAB species in pre-ensiled crops was far better detected by NGS. Likewise, good survival of Agrobacterium spp., Methylobacterium spp., and Sphingomonas spp. in IR and AL silages was demonstrated by NGS. The diversity of the microbiota described by principal coordinate analysis was similar between DGGE and NGS. Our finding that analysis of pre-ensiled crop microbiota did not help predict silage microbiota was true for both DGGE and NGS.

DOI： 10.1007/s00253-016-7900-2

Dietary casein and soy protein isolate modulate the effects of raffinose and fructooligosaccharides on the composition and fermentation of gut microbiota in rats Reviewed

Gaowa Bai, Kuikui Ni, Takeshi Tsuruta, Naoki Nishino

Journal of Food Science 81 ( 8 ) H2093 - H2098 2016.8

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

Although diet has an important influence on the composition of gut microbiota, the impact of dietary protein sources has only been studied to a minor extent. In this study, we examined the influence of different dietary protein sources regarding the effects of prebiotic oligosaccharides on the composition and metabolic activity of gut microbiota. Thirty female rats were fed casein and soy protein isolate with cellulose, raffinose (RAF), and fructooligosaccharides (FOS). Microbiota composition was examined by real-time qPCR and denaturing gradient gel electrophoresis. Dietary protein source affected cecum microbiota; acetic acid concentration and Lactobacillus spp. populations were greater with soy protein than with casein. Prebiotic oligosaccharides had distinctive effects on gut microbiota; RAF increased the acetic acid concentration and Bifidobacterium spp. populations, and FOS increased the butyric acid concentration regardless of the dietary protein. Likewise, Bifidobacterium sp., Collinsella sp., and Lactobacillus sp. were detected in microbiota of the rats fed RAF, and Bacteroides sp., Roseburia sp., and Blautia sp. were seen in microbiota of the rats fed FOS. Interactions between dietary proteins and prebiotic oligosaccharides were observed with Clostridium perfringens group populations and cecum IgA concentration. RAF and FOS decreased C. perfringens group populations in casein-fed rats, and the combination of soy protein and RAF substantially increased cecum IgA concentration. These results indicate that dietary proteins can differentially modulate the effects of prebiotic oligosaccharides on gut fermentation and microbiota, depending on the type of carbohydrate polymers involved.

DOI： 10.1111/1750-3841.13391

Lactic acid bacteria in total mixed ration silage containing soybean curd residue: their isolation, identification and ability to inhibit aerobic deterioration Reviewed

Li, Y., Wang, F., Nishino, N.

Asian-Australasian Journal of Animal Sciences 29 ( 4 ) 516 - 522 2016.4

Language：English Publishing type：Research paper (scientific journal) Publisher：ASIAN-AUSTRALASIAN ASSOC ANIMAL PRODUCTION SOC

We investigated the effects of the predominant lactic acid bacteria (LAB) on the fermentation characteristics and aerobic stability of total mixed ration (TMR) silage containing soybean curd residue (SC-TMR silage). The SC-TMR materials were ensiled in laboratory silos for 14 or 56 days. LAB predominant in SC-TMR silage were identified (Exp. 1). Lactobacillus fermentum (L. fermentum) and Streptococcus bovis (S. bovis) were found in the untreated materials, Leuconostoc pseudomesenteroides (L. pseudomesenteroides) in 14-day silage and Lactobacillus plantarum (L. plantarum) in all silages. Pediococcus acidilactici (P. acidilactici), Lactobacillus paracasei (L. paracasei), and Lactobacillus brevis (L. brevis) formed more than 90% of the isolates in 56-day silage. Italian ryegrass and whole crop maize were inoculated with P. acidilactici and L. brevis isolates and the fermentation and aerobic stability determined (Exp. 2). Inoculation with P. acidilactici and L. brevis alone or combined improved the fermentation products in ryegrass silage and markedly enhanced its aerobic stability. In maize silage, P. acidilactici and L. brevis inoculation caused no changes and suppressed deterioration when combined with increases in acetic acid content. The results indicate that P. acidilactici and L. brevis may produce a synergistic effect to inhibit SC-TMR silage deterioration. Further studies are needed to identify the inhibitory substances, which may be useful for developing potential antifungal agents.

DOI： 10.5713/ajas.15.0267

Identification and isolation of Lactobacillus fructivorans from wilted alfalfa silage with and without molasses Reviewed

B. Wu, N. Nishino

Journal of Applied Microbiology 120 ( 3 ) 543 - 551 2016.3

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

AimTo gain insights into the bacterial species associated with anaerobic storage and aerobic stability of alfalfa silage.
Methods and ResultsWilted alfalfa silage (498gdry matterkg(-1)) was prepared with and without the addition of molasses. Aerobic spoilage tests were conducted at 5, 10 and 60days after ensiling. The composition of fermentation products and the bacterial communities of silage were determined at 1, 3, 5 and 7days after silo opening. Silage without molasses had small amounts of lactic and acetic acids detectable at silo opening but resisted deterioration due to aerobic spoilage for at least 5days after opening. Resistance to aerobic deterioration in silage increased with the addition of molasses. The predominant bacterial species in molasses-added silage was Lactobacillus fructivorans, which was detected by denaturing gradient gel electrophoresis analysis. Different bacterial growth media were used for Lact.fructivorans isolation from alfalfa silage with added molasses: isolation was successful using liver infusion sake medium, but was unsuccessful when de Man, Rogosa and Sharpe medium was used.
ConclusionA nonconventional lactic acid bacterium (LAB) species may be involved in the high aerobic stability of alfalfa silage.
Significance and Impact of the StudyThe findings demonstrate that culture-independent microbiota analysis may be useful in the isolation and identification of nonconventional LAB species involved in fermentation and the aerobic stability of silage.

DOI： 10.1111/jam.13031

Fermentation and microbial population dynamics during the ensiling of native grass and subsequent exposure to air Reviewed

Qing Zhang, Baiyila Wu, Naoki Nishino, Xianguo Wang, Zhu Yu

Animal Science Journal 87 ( 3 ) 389 - 397 2016.3

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY

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 20days in LN silage and the pH decreased to 6.13 after 45days of ensiling. The temperature of the LN silage increased after approximately 30h 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.

DOI： 10.1111/asj.12427

Identification of bacteria in total mixed ration silage produced with and without crop silage as an ingredient Reviewed

Naoki Nishino, Yu Ogata, Hongyan Han, Yasunari Yamamoto

Animal Science Journal 86 ( 1 ) 45 - 50 2015.1

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

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.

DOI： 10.1111/asj.12234

Bacterial communities in alfalfa and corn silages produced in large-scale stack and bunker silos in China Reviewed

Baiyila Wu, Qing Zhang, Zhongkuan Liu, Zhu Yu, Naoki Nishino

Grassland Science 60 ( 4 ) 247 - 251 2014.12

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

Alfalfa silage (Al) and corn silage (Cs) samples were collected from two large-scale farms in China; one of these farms used stack silos and the other used bunker silos for silage production. Three samples were collected from the upper and lower layers of each silo. Thus, six samples were examined for each of Al-stack, Al-bunker, Cs-stack, and Cs-bunker. Bacterial community was determined by denaturing gradient gel electrophoresis (DGGE). Lactic acid was the main fermentation product in Al-stack, Cs-stack and Cs-bunker, whereas acetic acid and butyric acid were predominant in Al-bunker. Pediococcus pentosaceus and Clostridium sp. were common to four silage types. Lactobacillus acetotolerans and Lactobacilluspontis were detected specifically in Cs, whereas the lactic acid bacteria identified in Al were regarded as those usually found in silage. Differences between stack and bunker silos were apparent in Al with regard to fermentation products and DGGE band patterns. Differences between the upper and lower layers were observed in the fermentation products in Cs regardless of silo type, whereas variations in the DGGE band patterns of Cs-stack were detected.

DOI： 10.1111/grs.12063

A pilot examination of the fermentation products, aerobic stability and bacterial community of total mixed ration silage produced in Vietnam Reviewed

Tu Tran Thi Minh, Van Nguyen Huu, Naoki Nishino

Grassland Science 60 ( 1 ) 63 - 68 2014.3

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

To evaluate the fermentation characteristics and aerobic stability of total mixed ration (TMR) silage produced in the tropics, rice straw (Rs) and corn stover (Cst) were mixed with molasses (M) and other feeds as TMRs, which were then preserved as silage in Hue, Vietnam. The silages were opened after 4months, and the microbial counts, fermentation product levels and aerobic stability were determined. The bacterial community was assessed by denaturing gradient gel electrophoresis (DGGE). All the silages were well preserved, with lactic acid serving as the major preservative. However, the lactic acid content was low (5.99gkg(-1) dry matter [DM]) in the Rs-M silage, while the ethanol content was substantial (37.2gkg(-1)DM) in the Cst-M silage. Heating due to spoilage was observed at 115 and 81h after silo opening in the Cst-M and Cst-TMR silages, respectively, whereas no heating was observed for 7days after the opening of the Rs-M and Rs-TMR silages. Among the 14 bacterial strains identified in the silages by the DGGE analysis, only three were lactic acid bacteria. Lactobacillus brevis and Weissella paramesenteroides were detected in all silages, while W.cibaria was only detected in the Rs-M and Cst-M silages. In the aerobically unstable Cst-TMR silage, bands indicative of Acetobacter pasteurianus, Staphylococcus sp. and Streptomyces sp. were specifically observed. These results indicate that although desirable lactic acid fermentation can be expected in a TMR silage in a tropical environment, aerobic stability is lowered if Cst instead of Rs is used as the ingredient crop. The presence of A.pasteurianus in the air-tight laboratory silo was unusual, but it could account for the low aerobic stability of the Cst-TMR silage.

DOI： 10.1111/grs.12041

A survey of fermentation products and bacterial communities in corn silage produced in a bunker silo in China Reviewed

Chao Wang, Hongyan Han, Xueying Gu, Zhu Yu, Naoki Nishino

Animal Science Journal 85 ( 1 ) 32 - 36 2014.1

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

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 250g/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 (<5.0g/kg dry matter (DM)), two Wc silages had 1,2-propanediol levels >25g/kg DM. In contrast, 3 St silages had large amounts (>10g/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.

DOI： 10.1111/asj.12076

Effects of ensiling fermentation and aerobic deterioration on the bacterial community in italian ryegrass, guinea grass, and whole-crop maize silages stored at high moisture content Reviewed

Yanbing Li, Naoki Nishino

Asian-Australasian Journal of Animal Sciences 26 ( 9 ) 1304 - 1312 2013.9

Language：English Publishing type：Research paper (scientific journal)

The effects of storage period and aerobic deterioration on the bacterial community were examined in Italian ryegrass (IR), guinea grass (GG), and whole-crop maize (WM) silages. Direct-cut forages were stored in a laboratory silo for 3, 7, 14, 28, 56, and 120 d without any additives
live counts, content of fermentation products, and characteristics of the bacterial community were determined. 2,3-Butanediol, acetic acid, and lactic acid were the dominant fermentation products in the IR, GG, and WM silages, respectively. The acetic acid content increased as a result of prolonged ensiling, regardless of the type of silage crop, and the changes were distinctively visible from the beginning of GG ensiling. Pantoea agglomerans, Rahnella aquatilis, and Enterobacter sp. were the major bacteria in the IR silage, indicating that alcoholic fermentation may be due to the activity of enterobacteria. Staphylococcus sciuri and Bacillus pumilus were detected when IR silage was spoiled, whereas between aerobically stable and unstable silages, no differences were seen in the bacterial community at silo opening. Lactococcus lactis was a representative bacterium, although acetic acid was the major fermentation product in the GG silage. Lactobacillus plantarum, Lactobacillus brevis, and Morganella morganii were suggested to be associated with the increase in acetic acid due to prolonged storage. Enterobacter cloacae appeared when the GG silage was spoiled. In the WM silage, no distinctive changes due to prolonged ensiling were seen in the bacterial community. Throughout the ensiling, Weissella paramesenteroides, Weissella confusa, and Klebsiella pneumoniae were present in addition to L. plantarum, L. brevis, and L. lactis. Upon deterioration, Acetobacter pasteurianus, Klebsiella variicola, Enterobacter hormaechei, and Bacillus gibsonii were detected. These results demonstrate the diverse bacterial community that evolves during ensiling and aerobic spoilage of IR, GG, and WM silages. © 2013 by Asian-Australasian Journal of Animal Sciences.

DOI： 10.5713/ajas.2013.13175

Changes in the bacterial community and composition of fermentation products during ensiling of wilted Italian ryegrass and wilted guinea grass silages Reviewed

Yanbing Li, Naoki Nishino

Animal Science Journal 84 ( 8 ) 607 - 612 2013.8

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

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.

DOI： 10.1111/asj.12046

Effects of inoculation of Lactobacillus rhamnosus and Lactobacillus buchneri on fermentation, aerobic stability and microbial communities in whole crop corn silage Reviewed

Yanbing Li, Naoki Nishino

Grassland Science 57 ( 4 ) 184 - 191 2011.12

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

Whole crop corn silage may be prone to spoilage after exposure to air. To investigate control of fermentation and aerobic spoilage by using silage additives, Lactobacillus rhamnosus and Lactobacillus buchneri were added to whole crop corn silage, and shifts in microbial communities were examined. Bacterial and fungal communities were identified by denaturing gradient gel electrophoresis analysis at the time of opening the silo and after conducting aerobic deterioration tests. Inoculation of L. rhamnosus did not affect fermentation, whereas that of L. buchneri decreased the lactic acid content and increased the acetic acid content. Aerobic stability was enhanced in silage stored for a long period (120 days), with increases in the acetic acid content even without L. buchneri inoculation. A high aerobic stability was achieved with a shorter ensiling period (56 days) in L. buchneri-inoculated silage. Lactic acid bacteria (LAB) inoculation did not alter the indigenous bacterial community, and the inoculated species were detected only as additions. Lactobacillus brevis, Pediococcus parvulus, Weissella confusa and Klebsiella pneumoniae were found in both the pre-ensiled crop and the silage, whereas Weissella paramesenteroides, Lactobacillus plantarum and Lactobacillus lactis were seen exclusively after ensiling. Inoculation also had little effect on the fungal community during the fermentation process. Candida magnolia, Cryptococcus flavus and Candida intermedia were detected, both in the pre-ensiled crop and the silage, whereas Candida glabrata and Candida quercitrusa were identified exclusively in the silage. Marked changes were seen in the fungal community after aerobic spoilage; Saccharomyces exiguus, Saccharomyces martiniae, Pichia fermentans, Pichia deserticola and Pichia kudriavzevii appeared in the spoiled silage. These results indicated that LAB inoculation produces few changes in bacterial and fungal communities in corn silage and that substantial changes occur in the fungal community when the silage is spoiled.

DOI： 10.1111/j.1744-697X.2011.00226.x

Monitoring the bacterial community of maize silage stored in a bunker silo inoculated with Enterococcus faecium, Lactobacillus plantarum and Lactobacillus buchneri Reviewed

Y. Li, N. Nishino

Journal of Applied Microbiology 110 ( 6 ) 1561 - 1570 2011.6

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

Aims:
To monitor variations in the bacterial community and fermentation products of maize silage within and between bunker silos.
Methods and Results:
Silage samples were collected in 2008 and 2009 from three dairy farms, wherein the farmers arranged for a contractor to produce maize silage using bunker silos. Silage was prepared using a lactic acid bacteria (LAB) inoculant consisting of Enterococcus faecium, Lactobacillus plantarum and Lactobacillus buchneri. Eight samples were collected from each bunker silo; 4 'outer' and 4 'inner' samples were collected from near the top and the bottom of the silo. The dry matter, lactic acid, acetic acid, ethanol, 1-propanol and 1,2-propanediol contents differed between bunker silos in both sampling years. Higher acetic acid, 1-propanol and 1,2-propanediol contents were found in the bottom than the top layers in the 2008 samples, and higher lactic acid content was found in the top than the bottom layers in the 2009 samples. The bacterial community varied more between bunker silos than within a bunker silo in the 2008 samples, whereas differences between the top and the bottom layers were seen across bunker silos in the 2009 samples. The inoculated LAB were uniformly distributed, while several nonconventional silage bacteria were also detected. Lactobacillus acetotolerans, Lactobacillus panis and Acetobacter pasteurianus were detected in both years. Stenotrophomonas maltophilia was detected in the 2008 samples, and Lactobacillus reuteri, Acinetobacter sp. and Rahnella sp. were detected in the 2009 samples.
Conclusions:
Although differences were seen within and between bunker silos, the bacterial community may indicate a different relationship between bunker silos and sampling locations within a bunker silo from that indicated by the fermentation products.
Significance and Impact of the Study:
Analysis of bacterial community can help understand how diverse non-LAB and LAB species are involved in the ensiling process of bunker-made maize silage.

DOI： 10.1111/j.1365-2672.2011.05010.x

Chemical Changes during Ensilage and In sacco Degradation of Two Tropical Grasses: Rhodesgrass and Guineagrass Treated with Cell Wall-degrading Enzymes Reviewed

Yu Zhu, Naoki Nishino, Guo Xusheng

Asian-Australasian Journal of Animal Sciences 24 ( 2 ) 214 - 221 2011.2

Language：English Publishing type：Research paper (scientific journal) Publisher：ASIAN-AUSTRALASIAN ASSOC ANIMAL PRODUCTION SOC

Effects of the cell wall-degrading enzymes derived from Acremonium cellulolyticus and Trichoderma viride on the silage fermentation and in sacco degradation of tropical grasses i.e. rhodesgrass (Chloris gayana Kunth. cv. Callide) and guineagrass (Panicum maximum Jacq. cv. Natsukaze) were investigated in laboratory-scale experiments. These two grasses were either treated with or without the enzymes before ensiling. Untreated rhodesgrass produced acetate fermentation silage (lactate, 13.0 g kg(-1) DM; acetate, 38.7 g kg(-1) DM) with high final pH value and NH(3)-N content (5.84 and 215 g kg(-1) DM). Addition of enzymes significantly increased (p<0.01) the lactate production (lactate, 45.6; acetate, 34.0 g kg(-1) DM) and decreased (p<0.01) the pH and NH(3)-N (4.80 and 154 g kg(-1) DM) in the ensiled forages when compared with the control silages. Untreated guineagrass was successfully preserved with a high lactate proportion (lactate, 45.5; acetate, 24.1 g kg(-1) DM), and the addition of enzymes further enhanced the desirable fermentation (lactate, 57.5 g kg(-1) DM: acetate, 19.4 g kg(-1) DM). The content of NDF was lowered (p<0.05) by enzymes in both silages, but the extent appeared greater in the enzyme-treated rhodesgrass (rhodesgrass, 48 g kg(-1) DM; guineagrass, 21 g kg(-1) DM). Changes in the kinetics of in sacco degradation showed that enzyme treatment increased (p<0.01) the rapidly degradable DM (rhodesgrass, 299 vs. 362 g kg(-1) DM; guineagrass, 324 vs. 343 g kg(-1) DM) but did not influence the potential degradation, lag time and degradation rate of DM and NDF in the two silages.

Presence of sourdough lactic acid bacteria in commercial total mixed ration silage as revealed by denaturing gradient gel electrophoresis analysis

C. Wang, N. Nishino

LETTERS IN APPLIED MICROBIOLOGY 51 ( 4 ) 436 - 442 2010.10

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

Aims:
To characterize the bacterial communities in commercial total mixed ration (TMR) silage, which is known to have a long bunk life after silo opening.
Methods and Results:
Samples were collected from four factories that produce TMR silage according to their own recipes. Three factories were sampled three times at 1-month intervals during the summer to characterize the differences between factories; one factory was sampled 12 times, three samples each during the summer, autumn, winter and spring, to determine seasonal changes. Bacterial communities were determined by culture-independent denaturing gradient gel electrophoresis. All silages contained lactic acid as the predominant acid, and the contents appeared stable regardless of factories and product seasons. Acetic acid and 1-propanol contents were different between factories and indicated seasonal changes, with increases in warm seasons compared to cool seasons. Both differences and similarities existed among the bacterial communities from each factory and product season. Lactobacillus parabuchneri was found in the products from three of four factories. Various sourdough lactic acid bacteria (LAB) were identified in commercial TMR silage; Lactobacillus panis, Lactobacillus hammesii, Lactobacillus mindensis, Lactobacillus pontis, Lactobacillus frumenti and Lactobacillus farciminis were detected in many products. Moreover, changes owing to product season were distinctive, and Lact. pontis and Lact. frumenti became detectable in summer products.
Conclusion:
Sourdough LAB are involved in the ensiling of commercial TMR silage. Silage bacterial communities vary more by season than by factory. The LAB species Lact. parabuchneri was detected in the TMR silage but may not be essential to the product's long bunk life after silo opening.
Significance and Impact of the Study:
Commercial TMR silage resembles sourdough with respect to bacterial communities and long shelf life. The roles of sourdough LAB in the ensiling process and aerobic stability are worth examining.

DOI： 10.1111/j.1472-765X.2010.02915.x

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.

Changes in Soluble Sugars and Fermentation Products of TMR Silages Prepared with High and Low Malt Type Brewer's Grains Reviewed

NISHINO Naoki, HARADA Hiroaki, SAKAGUCHI Ei

Japanese Journal of Grassland Science 49 ( 4 ) 367 - 372 2003

Authorship：Corresponding author Language：Japanese Publisher：Japanese Society of Grassland Science

Two types of brewer's grains (BG), derived from high and low malt brewing, were mixed with various dry feeds (BG : beet pulp : oats hay : wheat bran : molasses=5 : 2 : 1 : 1 : 1 on a fresh weight basis), and ensiled in polyethylene bottles to prepare a total mixed ration (TMR silage). Silos were opened after 5, 20 and 40 d of storage ; the composition of soluble sugars and the profile of fermentation products were determined. Experiment was duplicated using different lots of BG (Exp. 1 & 2). Regardless of the type of BG, lactic acid production occurred and large numbers of lactic acid bacteria were found at 12 h after BG production. Principal sugars were maltose and raffinose in both BG, whilst the amount was higher in low malt type BG. When TMR silage was made, the main sugars altered to fructose, glucose and sucrose due to added feeds, and the differences in sugar contents diminished. Lactic acid (Exp. 1) or ethanol (Exp. 2) was the major fermentation product in TMR silage, and this property was not influenced by the type of BG used. Mannitol accumulated in lactate silages as storage prolonged, indicating that heterolactic fermentation was enhanced even when lactate silage was obtained. Accelerated ethanol production was found in ethanol silage from the beginning of storage ; the highest content (75gkg^<-1> DM) was given in TMR silage prepared with low malt type BG. Propionic and butyric acids were almost undetectable and ammonia N accounted for less than 60gkg^<-1> N in any of the silages, suggesting that TMR silage can be considered "well preserved" regardless of lactate and ethanol fermentation. Significant differences due to high and low malt type BG were found in the fermentation of TMR silage ; however, the production lot had much greater effects in the two experiments.

DOI： 10.14941/grass.49.367

Other Link： http://agriknowledge.affrc.go.jp/RN/2010682300

Response of intestinal proteinase activities to the feeding of isolated winged bean (Psophocarpus tetragonolobus) and soya bean (Glycine max) trypsin inhibitors in rats Reviewed

N Nishino, M Taniguchi, E Sakaguchi

Journal of the Science of Food and Agriculture 81 ( 11 ) 1055 - 1059 2001.9

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

The antinutritional activities of trypsin inhibitors (TIs) were compared between winged beans (Psophocarpus tetragonolobus) and soya beans (Glycine max). The inhibitors of the two beans were isolated by trypsin-bound Sepharose 4B, and 50 mg of lyophilised powders were intubated intragastrically into 24 h fasted rats. The activities of trypsin and chymotrypsin were compared after 30, 60 and 180 min in the washings of the upper, middle and lower parts of the small intestine. The elution profiles of TI and non-TI compounds in the affinity chromatography were similar in the two beans, and the antitryptic activities were concentrated 5.5 and 6.2 times (based on specific activity) for winged beans and soya beans respectively. Regardless of the TI fed to rats, trypsin activity in the upper intestine was suppressed to almost undetectable levels at 30 and 60 min after intubation. The activities in the middle and lower intestines were also substantially lowered when rats were fed winged bean TI, and significant differences were detected at 30 and 60 min after intubation when compared with rats fed soya bean TI. However, at ISO min after feeding, no differences were found in the trypsin activity in any gut segments. Similar inhibitory properties of isolated TIs were observed in chymotrypsin activities in the small intestine. The results suggest that winged bean TI may have greater inhibitory activity on the intestinal proteinase compared with soya bean TI. (C) 2001 Society of Chemical Industry.

DOI： 10.1002/jsfa.891

Disappearance of Maillard reaction products during ensilage and rumen fermentation in vitro Reviewed

N Nishino, S Uchida

Journal of the Science of Food and Agriculture 81 ( 2 ) 275 - 280 2001.1

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

The disappearance of Maillard reaction products (MRPs) during ensilage and rumen fermentation was studied. MRPs were prepared by heating mixtures of D-glucose and glycine (GG) or D-xylose and glycine (XG); lyophilised powder (MW > 1000) was subjected to anaerobic incubation for 24h with lucerne juice or buffered rumen fluid. Changes in MRPs were assessed by comparing gel filtration profiles before and after incubation, and the proportion of disappeared MRPs was calculated based on the area under the curve. A portion of MRPs, particularly high-MW fractions, disappeared when incubated with lucerne juice, and the extent was greater (P < 0.01) in XG than in GG. The disappearance of GG was increased (P < 0.01) when ensilage was fortified with added glucose, while that of XG was unaffected with or without the fermentable substrate. Rumen fermentation had little impact on GG and XG, whereas a portion of MRPs disappeared when microbial activity was enhanced by adding lucerne hay to the media. Volatile fatty acid production was unaffected by MRPs in either ensilage or rumen fermentation. These results suggest that the effect on MRPs of anaerobic fermentation may be different between ensilage and rumen digestion. Significant amounts of MRPs would enter the intestine when ruminants are fed hay-based diets rather than silage-based diets. (C) 2000 Society of Chemical Industry.

DOI： 10.1002/1097-0010(20010115)81:2<275::AID-JSFA814>3.0.CO;2-L

Silage Making and Utilization of High Moisture By-products(Part 1) : 3. Ensiling Characteristics of Brewer's Grain Left after the Production of Beer and Happo-shu(Low Malt Beer)<Special Feature> Reviewed

NISHINO Naoki, HARADA Hiroaki, SAKAGUCHI Ei

Japanese Journal of Grassland Science 47 ( 3 ) 318 - 322 2001

Fermentation of Rhodesgrass and Guineagrass Silages with or without Wilting and Added Cell Wall Degrading Enzymes Reviewed

ZHU Yu, NISHINO Naoki, UCHIDA Senji

Japanese Journal of Grassland Science 46 ( 1 ) 22 - 27 2000

Authorship：Corresponding author Language：English Publisher：Japanese Society of Grassland Science

The effects of wilting and added enzymes on fermentation and in vitro digestion of rhodesgrass and guineagrass silages were examined. First cuts of the two forages were ensiled in laboratory silos directly or after being wilted for 4 h with or without cell wall degrading enzymes (0.2 g kg^<-1>). Silos were opened after 45 d and the chemical composition and the fermentation quality were determined. The silages were subjected to in vitro rumen incubation and the degradation of NDF was determined. Without enzymes rhodesgrass silage failed to achieve lactate type fermentation, while guineagrass silage had high proportion of lactate even without any of the treatments. Wilting inhibited the organic acid production but decreased the proportion of lactate in rhodesgrass silage. Use of enzymes improved the preservation of both the silages, as evidenced by the increase of lactate and the decrease of pH value and NH3-N. The contents of NDF and ADF were reduced by enzyme treatment, but the reduction was considerably suppressed when forages were wilted before ensiling. In vitro degradation of NDF was low in direct-cut rather than wilted silage. Addition of enzymes also lowered the degradation, but this effect was restrictedly found in guineagrass silage incubated for 72h. These results indicate that although added cell wall degrading enzymes can promise an improved fermentation of tropical grass silage, the benefit would be less pronounced when treated with low moisture material.

DOI： 10.14941/grass.46.22_1

Other Link： http://agriknowledge.affrc.go.jp/RN/2010611429

Ensiling characteristics and ruminal degradation of Italian ryegrass and lucerne silages treated with cell wall-degrading enzymes Reviewed

Y Zhu, N Nishino, Y Kishida, S Uchida

JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 79 ( 14 ) 1987 - 1992 1999.11

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

Two experiments were carried out to investigate the effect of added cell wall-degrading enzymes at ensiling on the fermentation and in situ degradation of grass and legume silages. Primary growths of Italian ryegrass (Lolium multiflorum Lam) and lucerne (Medicago sativa L) were wilted and ensiled in laboratory-scale silos with or without enzymes. Silages were opened at 2, 5, 15 and 45 days after storage; the fermentation quality and the contents of cell wall components (NDF, ADF, ADL) were determined. The 45 day silages were subjected to in situ incubation experiments, and the kinetics of DM and NDF degradation was estimated. The enzyme treatment enhanced the lactic acid production (P < 0.01) and reduced the pH value (P < 0.01) of both Italian ryegrass and lucerne silages. The contents of cell wall components, however, were not affected by enzymes, except for NDF of Italian ryegrass silage. The in situ incubation experiments revealed that added enzymes increased the rapidly degradable DM (P < 0.01) and appeared to decrease the rate of degradation of DIM and NDF of Italian ryegrass silage. The rapidly degradable DM was not altered when lucerne was treated, but the rate of degradation of NDF was significantly reduced (P < 0.05). These results suggest that although added cell wall-degrading enzymes could improve the preservation of grass and legume silages, the effects on ruminal degradation may be different according to the herbage treated. Enzymatic hydrolysis during ensilage may be restricted to easily digestible cell walls, leaving relatively less digestible components that would be retained in treated silages. (C) 1999 Society of Chemical Industry.

DOI： 10.1002/(SICI)1097-0010(199911)79:14<1987::AID-JSFA466>3.0.CO;2-J

Laboratory evaluation of previously fermented juice as a fermentation stimulant for lucerne silage Reviewed

N Nishino, S Uchida

JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 79 ( 10 ) 1285 - 1288 1999.7

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

The potential use of forage juice anaerobically stored for 2 days (previously fermented juice, PFJ) as a silage additive was investigated. Lucerne (450 g) was ensiled in laboratory-scale silos (900 ml capacity) with or without sucrose (20 g kg(-1)) or PFJ (2.5 ml kg(-1)). Silages treated with both additives were also made and fermentation quality, in vitro dry matter digestibility and in vitro ruminal NH3 production were determined. The control silage showed very poor quality with high butyrate and NH3-N concentrations. Addition of sucrose significantly increased lactate but failed to inhibit the deterioration by clostridial fermentation. Use of PFJ further increased lactate and greatly reduced butyrate and NH3-N production during storage. The combination of sucrose and PFJ was more effective in improving fermentation than either additive on its own. In vitro digestibility appeared to be higher when lucerne was well preserved. Ruminal NH3 production in vitro digestibility appeared to be higher when lucerne was well preserved. Ruminal NH3 production in vitro was decreased in silages with sucrose and PFJ added at ensiling. This result indicates that use of PFJ can be a promising way to stimulate lactate production in some difficult direct cut silage. The potential of improving fermentation may be comparable to conventional sugar additives. (C) 1999 Society of Chemical Industry.

DOI： 10.1002/(SICI)1097-0010(19990715)79:10<1285::AID-JSFA362>3.0.CO;2-M

A note on the chemical changes associated with nonenzymatic browning of forages Reviewed

Nishino, N, Touno, E, Uchida, S

Grassland Science 44 ( 1 ) 86 - 89 1998.1

Effects of cell wall degrading enzymes and lactic acid bacteria on the fermentation of rhodesgrass (Chloris gayana Kunth.) silage stored at various ambient temperature Reviewed

NISHINO N

Grassland Science 44 199 - 203 1998

Effects of extraction and reconstitution of ryegrass juice on fermentation, digestion and in situ degradation of pressed cake silage Reviewed

Naoki Nishino, Keiko Miyase, Mitsuaki Ohshima, Hiro-Omi Yokota

Journal of the Science of Food and Agriculture 75 ( 2 ) 161 - 166 1997.10

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

Primary growth of Italian ryegrass was fractionated into green juice and pressed cake (PC). PC was ensiled either directly or after reconstitution with the juice which was spray dried and then added to water before ensiling (RPC). This process was aimed at distinguishing between chemical (removal of nutrients) and physical (mechanical disintegration) effects of fractionation. Two additional wilted silages were prepared from long cut or chopped Italian ryegrass (WL and WC, respectively). The fermentative quality, digestion and in situ degradation of dry matter for the four silages were compared. WC, RPC and PC silages indicated lactate-rich fermentation while WL silage had a comparatively low lactate content. PC silage had a lower (P &lt
0.05) pH value with lower (P &lt
0.05) amounts of organic acids compared with WC and RPC silage, suggesting that removal of buffering components with the juice had greater effects than shredding on fermentation of PC silage. The digestibility of protein was lower (P &lt
0.05) in PC than WL silage but the reduction was compensated for in RPC silage. Ruminal concentrations of total volatile fatty acids were the highest (P &lt
0.05) when goats fed WL silage while those on the other silages had similar concentrations. Ruminal pH was lower (P &lt
0.05) in goats fed PC than WC silage and the difference was not fully reversed by reconstituting the juice to PC. Juice extraction decreased (P &lt
0.05) the soluble fraction and the rate of disappearance of dry matter in the rumen but the potential degradability was not different among treatments. These results suggest that both chemical and physical changes due to the fractionation affect the characteristics of digestion of PC silage while the removal of nutrients has a major role in the modified silage fermentation.

DOI： 10.1002/(SICI)1097-0010(199710)75:2<161::AID-JSFA854>3.0.CO;2-Q

Ruminal and whole tract digestion of long cut, short cut and pressed cake hay prepared from Italian ryegrass by goats Reviewed

Nishino, N, Miyase, K, Ohshima, M, Yokota, H

Grassland Science 43 ( 2 ) 83 - 87 1997

Authorship：Lead author Language：English Publisher：Japanese Society of Grassland Science

From a winter harvest of autumn-sown Italian ryegrass, long cut, short cut and pressed cake hay were prepared. The three hays were solely fed to goats equipped with rumen cannulae and the digestibility of nutrients, rumen fluid characteristics and in situ disappearance of dry matter were determined. The content of fibrous components increased during hay-making and the increased proportion was the highest in short cut hay and the lowest in pressed cake hay. The digestibility of pressed cake hay was similar to those of the other hays except for protein which was lower in pressed cake hay. The digestibilities of NDF and hemicellulose of short cut hay were significantly higher than those of long cut hay. Goats given pressed cake hay showed lower ruminal pH than those given short cut hay while maintaining less concentrations of volatile fatty acids after feeding. Ruminal degradation characteristics of dry matter revealed that short cut hay had less soluble fraction but higher rate of degradation compared with long cut hay. The pressing of macerated Italian ryegrass further decreased the soluble fraction but potential degradability was not lowered despite of the removal of highly digestible nutrients. The rate of degradation was similar between long cut and pressed cake hay. These results indicate that the chopping before hay-making would alter the characteristics of ruminal digestion of Italian ryegrass while not affecting the whole tract digestibility. Pressed cake hay was comparable to long cut hay in terms of digestibility and rate of degradation in the rumen.

DOI： 10.14941/grass.43.83

Other Link： http://agriknowledge.affrc.go.jp/RN/2010560580

Changes in nitrogenous compounds and rates of in situ N loss of soybean meal treated with sodium hydroxide or heat Reviewed

N Nishino, Y Masaki, S Uchida

Journal of Agricultural and Food Chemistry 44 ( 9 ) 2667 - 2671 1996.9

Authorship：Lead author,　Corresponding author Language：English Publishing type：Research paper (scientific journal) Publisher：AMER CHEMICAL SOC

Rates of in situ N loss of soybean meal (SBM) were altered by various combinations of sodium hydroxide and heat treatment; some nitrogenous compounds involving the modified proteolysis were investigated. The amount of lysinoalanine (LAL), neutral detergent insoluble nitrogen, 1-fluoro-2,4-dinitrobenzene reactive lysine, and reducing power of SBM were linearly related (P < 0.01) to rate of in situ N loss with correlation coefficients of -0.847, -0.835, 0.793, and -0.846, respectively. The results indicate that the LAL formation and the Maillard reaction are related to change in susceptibility of SBM protein to microbial degradation in the rumen.

DOI： 10.1021/jf9602481

Nitrogen utilization of goats fed silage with or without supplements having different rumen degradability Reviewed

Nishino, N, Miyase, K, Ohshima, M, Yokota, H

Grassland Science 41 ( 3 ) 202 - 206 1995

Authorship：Lead author Language：English Publisher：Japanese Society of Grassland Science

Effects of feeding corn, barley and sugar beet pulp to silage based diet on the nitrogen (N) utilization of goats were studied. These supplements were selected due to having the difference in rumen degradation. Four rumen cannulated goats were fed the diets consisting of wilted ryegrass (Lolium multiftorum Lam) silage and the supplements (4 : 1 on dry matter (DM) basis). Silage was solely used for the control diet and N intake was adjusted by urea added to the diets including supplements. The wilted silage showed the pH value 4.72 and relatively high NH3-N concentration (16.7%/total-N). The soluble fraction of DM in the rumen was 58.0, 26.8 and -1.25%, and the rate of degradation was 0.042, 0.107 and 0.099h^<-1>, for corn, barley and sugar beet pulp, respectively. Dietary inclusion of corn significantly increased DM and organic matter digestibility, and that of barley increased hemicellulose digestibility compared with the control diet. Urinary excretion of allantoin increased when goats fed the supplements, but the difference failed to be significant. Ruminal concentration of NH_3-N and retention of N showed no significant difference among the treatments, but feeding barley tended to lose more N compared with the other supplements in this study.

DOI： 10.14941/grass.41.202

Other Link： http://agriknowledge.affrc.go.jp/RN/2010532124

Formation of lysinoalanine following alkaline processing of soya bean meal in relation to the degradability of protein in the rumen Reviewed

N. Nishino, S. Uchida, M. Ohshima

Journal of the Science of Food and Agriculture 68 ( 1 ) 59 - 64 1995

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

Effects of sodium hydroxide (NaOH) treatment on the formation of lysinoalanine (LAL) and on in‐situ and in‐vivo utilisation of soya bean meal (SBM) protein were investigated. Defatted SBM was sprayed with 0 (water), 25, 50 and 100 g NaOH (kg−1 dry matter), dried at 80°C for 2 h and then fed to four rumen cannulated goats in a 4 × 4 Latin square as a mixed diet (9: 1 w/w) of sudangrass hay and the SBM. LAL was not detected in non‐alkaline SBM, but the cross‐linked amino acid increased according to the level of NaOH addition. The maximum concentration of LAL was 1.26 g per 16 g nitrogen when the highest level of NaOH was applied. The NaOH treatment did not alter the digestibility or the retention of nitrogen of the mixed diet. Addition of NaOH at 25 and 50 g (kg−1 dry matter) lowered the solubility of protein, while the highest level of treatment had little effect. Lag time and rate of degradation of protein decreased due to the increasing level in NaOH treatment. A significant correlation (r = —0.860, P &lt
0.01) between LAL content and rate of protein degradation suggested that the formation of cross‐linked amino acid would render protein more resistant to microbial degradation in the rumen. Copyright © 1995 John Wiley &amp
Sons, Ltd

DOI： 10.1002/jsfa.2740680110

Influence of conservation method on the fiber digestion of Italian ryegrass by goats Reviewed

Nishino, N, Wakita, Y, Uchida, S

Animal Science and Technology 65 ( 8 ) 709 - 715 1994.8

Authorship：Lead author Language：Japanese Publisher：Japanese Society of Animal Science

Effect of preservation as hay and silage on the fiber digestion of Italian ryegrass was investigated. From the same harvest at heading stage, field dried hay and wilted silage were prepared. These forages were restrictedly fed to four rumen cannulated goats as a diet consisting of forage-soybean meal (9:1) mixture to determine nutrient digestibility, nitrogen balance, rumen degradability of DM and NDF, particle-bound carboxymethylcellulase (CMCase) activity, and rate of passage of liquid (Co-EDTA) and solid digesta (Cr-CWC). The wilted silage (DM 30.8%) showed a relatively high pH value 4.75 and a butyrate-rich fermentation. The hay showed higher NDF, NDIN and ADIN contents than silage, suggesting some protein became associated with fibrous components during hay-making. All the digestibility of silage diet was higher than those of hay diet, while significant difference could not be detected in fiber digestibility. When goats were fed silage, they retained more nitrogen than those on hay. The silage had more rumen soluble (5.5 and 0.0%) and less degradable (71.4 and 74.4%) NDF compared with hay. The rate of NDF degradation was lower in silage than hay (3.2 and 4.2%h^-1), which may partly be ascribed to lower CMCase activity of silage. No significant difference was observed in the rate of passage of either liquid or feed particles. These results indicate that there is considerable difference as energy source in the fibrous components between hay and silage as well as a marked difference of soluble carbohydrate and volatile fatty acid contents.

DOI： 10.2508/chikusan.65.709

Other Link： https://jlc.jst.go.jp/DN/JALC/00367137654?from=CiNii

Effect of sodium hydroxide or ammonia treatment on digestion and rumen degradation of alfalfa silage in goats Reviewed

Nishino, N, Uchida, S, Ohshima, M

Journal of Japanese Society of Grassland Science 39 ( 4 ) 437 - 445 1994

Authorship：Lead author Language：English Publisher：Japanese Society of Grassland Science

Three rumen cannulated goats were used to investigate the effect of NaOH and NH_3 treatment (2.74% DM) at ensiling on digestion, nitrogen utilization and in situ protein degradation of low moisture alfalfa silage. Wilted alfalfa was sprayed with water (control), NaOH or NH_3 solution, and fed to goats after 6 months storage in 3×3 Latin square design. Both alkali treatments increased the pH value, NH_3-N and butyric acid of the silage, but NH_3-treated silage preserved as much amino acids as control silage. The two alkali treatments could not improve digestibilities of fibrous components (NDF, ADF and cellulose), and NaOH addition significantly reduced the digestibility of crude protein. The NaOH treatment decreased the rate of N degradation in the rumen, while solubility and potential degradability was not affected by the treatment. The NH_3 addition increased ruminal soluble nitrogen in the silage, but rate of N degradation was similar to that of control silage. These results suggest that NaOH treatment would make protein less susceptible to microbial degradation in the rumen despite of deteriorating the fermentation quality of the silage.

DOI： 10.14941/grass.39.437

Other Link： http://agriknowledge.affrc.go.jp/RN/2010510133

A note on the fermentation characteristics and rumen degradation of low moisture alfalfa silage treated with sodium hydroxide or ammonia Reviewed

N. Nishino, M. Ohshima, S. Uchida

Animal Feed Science and Technology 47 ( 3-4 ) 297 - 304 1994

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

The effects of alkali treatment on the fermentation characteristics and rumen degradation of low moisture alfalfa silage were investigated. First crop alfalfa was wilted and ensiled, either directly or after being sprayed with NaOH or NH3 solution (1.72% dry matter (DM)). The herbages were stored in laboratory silos and opened at 1, 2, 7, 14, 28 and 56 days after storage. Each NaOH and HN3 addition increased the pH value and delayed lactic acid production of the silage. Both alkali treatments decreased proteolysis during ensilage, and NaOH-treated silage showed the highest protein (hot water insoluble nitrogen) content. Nitrogen solubility in a mineral buffer solution was decreased by the alkali treatments, and NaOH-treated silage showed the lowest solubility. Degradation characteristics of DM and N in the rumen were determined by nylon bag incubation of 56th day silage samples with two rumen fistulated goats. The NaOH treatment significantly decreased the soluble N fraction (63.7%, 54.7% and 60.6% for untreated, NaOH-treated and NH3-treated silage, respectively) but did not influence the extent of degradation. The treatment tended to decrease the rate of N degradation. In contrast, the NH3 treatment significantly increased the rate of DM degradation (0.158 h-1, 0.151 h-1 and 0.182 h-1 for untreated, NaOH-treated and NH3-treated silage, respectively) without any great changes in the proportions of soluble and degradable fraction. © 1994.

DOI： 10.1016/0377-8401(94)90132-5

Ruminal degradation of alfalfa protein as influenced by sodium hydroxide and heat treatment Reviewed

Nishino, N., Uchida, S., Ohshima, M.

Animal Feed Science and Technology 48 ( 1-2 ) 131 - 141 1994

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

Effects of sodium hydroxide (NaOH) and heat treatment of alfalfa on the degradation of protein in the rumen was investigated in relation to the changes of nitrogenous compounds. Fresh alfalfa was treated with 0, 1.16, 2.32 or 4.64 g NaOH per 100 g dry matter (DM) and then subjected to freeze-drying, or oven-drying for 24 h at 60 or 120°C. Both NaOH and heat treatment produced lysinoalanine at increasing rate with level of treatment to reach a maximum value of 238 mg per 100 g protein. Both treatments also increased neutral detergent insoluble nitrogen (NDIN) according to the increase in level of treatment. Acid detergent insoluble nitrogen (ADIN) was not affected by NaOH treatment but was significantly increased by heating at 120°C. Both NaOH and heat treatment modified protein into being less soluble and more slowly degradable in the rumen, but heating showed greater effects than NaOH treatment. The proportion of potentially degradable protein was little influenced by the treatments. Both NaOH and heat treatment decreased the rate of protein degradation, but significant differences were detected only with the highest level of treatment (4.64 g NaOH per 100 g DM or 120°C). Correlation coefficients of lysinoalanine, NDIN and ADIN content with rate of protein degradation in the rumen were -0.716, -0.950 and -0.834, respectively. These results suggest that lysinoalanine formation in protein by NaOH and heat treatment can make protein less susceptible to microbial degradation in the rumen, while protein-carbohydrate reaction enhanced the effects on protein degradation in this study. © 1994.

DOI： 10.1016/0377-8401(94)90117-1

Digestion of alkali-treated alfalfa silage by goats Reviewed

Nishino, N, Miyase, K, Ohshima, M, Yokota, H

Asian-Australasian Journal of Animal Sciences 6 5 - 11 1993.1

Nutritive value of silages prepared from fiber and liquid residues after the separation of leaf nutrient concentrate from Italian ryegrass in goats Reviewed

Reddy, G.U, Ohshima, M, Nishino, S

Asian-Australasian Journal of Animal Sciences 5 249 - 256 1992

Effect of addition of intact or alkalized lucerne juice at ensiling on the nutritive value of rice straw silage Reviewed

Nishino, N, Ohshima, M, Yokota, H

Asian-Australasian Journal of Animal Sciences 5 487 - 494 1992

Nutritive value of rice straw ensiled with intact or alkalized Italian ryegrass green juice Reviewed

Nishino, N, Ohshima, M, Yokota, H

Journal of Japanese Society of Grassland Science 37 ( 2 ) 203 - 212 1991

Authorship：Lead author Language：English Publisher：Japanese Society of Grassland Science

Freshly harvested Italian ryegrass was disintegrated with a herbage crusher and pressed out green juice (GJ) with a screw press. The GJ was used as a protein supplement source for rice straw (RS). Rice straw was mixed with intact GJ (GJ・RS) and with NaOH or NH_3 (4% of RS dry matter) treated GJ (GJ・NaOH・RS or GJ・NH_3・RS) at 3:7 ratio on fresh weight basis and ensiled into polyethylene bags. Rice straw ensiled with water was also prepared as the control (W・RS). Crude protein content of RS was increased from 5.0 to 7.9% in dry matter basis by mixing intact or NaOH treated GJ and further increased to over 25% by NH_3 treated GJ. The control W・RS silage contained a trace amount of lactic acid and showed a butyrate type fermentation. The addition of intact GJ to RS significantly increased the lactic acid content, and reduced the butyric acid content, NH_3-N/TN and the pH value of the silage. In contrast to this, the addition of NaOH or NH_3 treated GJ increased the pH value, the butyric acid content and NH_3-N/TN of the silage, and GJ・NH_3・RS silage showed the lowest chemical quality. In the feeding trial, four castrated male goats were allocated in a 4×4 Latin-square and fed the 3:1 (DM basis) mixed ration of RS silage and wheat bran. Besides them, urea was supplemented to W・RS silage at the time of feeding to adjust the nitrogen intake except for that from NH_3 treated silage. The addition of intact GJ to RS at ensiling had little advantage to feeding urea with W・RS silage in respect to nutrient digestibilities but slightly improved nitrogen balance of goats. Compared with goats fed W・RS and GJ・RS silages, those fed GJ・NaOH・RS silage showed significantly higher fiber digestibilities, less urinary nitrogen excretion and more nitrogen retention with the reduction of ruminal NH_3-N and blood urea nitrogen concentrations. The goats fed GJ・NH_3・RS silage showed the highest nutrient digestibilities and nitrogen retention. These results suggest that the alkali treatments increased energy supply from fiber and consequently increased nitrogen uptake of rumen microbes. The disadvantage of feeding GJ・NaOH・RS compared with GJ・NH_3・RS was probably due to the insufficient nitrogen supply to rumen microbes for the maximum fiber digestion and protein synthesis.

DOI： 10.14941/grass.37.203

Other Link： http://agriknowledge.affrc.go.jp/RN/2010480737

Ruminal acid concentrations of goats fed hays and silages prepared from Italian ryegrass and its pressed cake Reviewed

Ohshima, M, Miyase, K, Nishino, N, Okajima, T, Yokota, H

Asian-Australasian Journal of Animal Sciences 4 59 - 65 1991

　 PREV - NEXT 　

1