• Journal of The Korean Society of Grassland and Forage Science
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• v.41 no.4
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• pp.302-307
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• 2021

In the current study, lactic lactic acid bacteria (LAB) Lactobacillus plantarum and Pediococcus pentosaceus were used as a mixed additive for the production of Orchardgrass silage by ensiled method and nutritional change fermentation ability and microbial content of experimental silages. The addition of LAB to Orchardgrass during ensiling process rapidly reduced the pH of the silages than the non-inoculated silages. In addition, the lactic and acetic acid content of silage was increased by LAB strains than the non-inoculated silages whereas butyric acid content was reduced in silage treated with LAB. A microbiological study revealed that higher LAB but lower yeast counts were observed in inoculated silages compared to non-inoculated silage. Overall data suggested that the addition of LAB stains could have ability to induce the fermentation process and improve the silage quality via increasing lactic acid and decreasing undesirable microbes.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.6
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• pp.788-796
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• 2017

Objective: In order to improve fermentation quality of natural grasses, their silage preparation and fermentation quality in meadow steppe (MS) and typical steppe (TS) were studied. Methods: The small-scale silages and round bale silages of mixed natural grasses in both steppes were prepared using the commercial lactic acid bacteria (LAB) inoculants Chikuso-1 (CH, Lactobacillus plantarum) and cellulase enzyme (AC, Acremonium cellulase) as additives. Results: MS and TS contained 33 and 9 species of natural grasses, respectively. Stipa baicalensis in MS and Stipa grandi in TS were the dominant grasses with the highest dry matter (DM) yield. The crude protein (CP), neutral detergent fiber and water-soluble carbohydrate of the mixed natural grasses in both steppes were 8.02% to 9.03%, 66.75% to 69.47%, and 2.02% to 2.20% on a DM basis, respectively. All silages treated with LAB and cellulase were well preserved with lower pH, butyric acid and ammonia-N content, and higher lactic acid and CP content than those of control in four kinds of silages. Compared with CH- or AC-treated silages, the CH+ AC-treated silages had higher lactic acid content. Conclusion: The results confirmed that combination with LAB and cellulase may result in beneficial effects by improving the natural grass silage fermentation in both grasslands.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.8
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• pp.1136-1144
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• 2016

Leucaena silage was supplemented with different levels of molasses and urea to study its nutritive value and in vitro rumen fermentation efficiency. The ensiling study was randomly assigned according to a $3{\times}3$ factorial arrangement in which the first factor was molasses (M) supplement at 0%, 1%, and 2% of crop dry matter (DM) and the second was urea (U) supplement as 0%, 0.5%, and 1% of the crop DM, respectively. After 28 days of ensiling, the silage samples were collected and analyzed for chemical composition. All the nine Leucaena silages were kept for study of rumen fermentation efficiency using in vitro gas production techniques. The present result shows that supplementation of U or M did not affect DM, organic matter, neutral detergent fiber, and acid detergent fiber content in the silage. However, increasing level of U supplementation increased crude protein content while M level did not show any effect. Moreover, the combination of U and M supplement decreased the content of mimosine concentration especially with M2U1 (molasses 2% and urea 1%) silage. The result of the in vitro study shows that gas production kinetics, cumulation gas at 96 h and in vitro true digestibility increased with the increasing level of U and M supplementation especially in the combination treatments. Supplementation of M and U resulted in increasing propionic acid and total volatile fatty acid whereas, acetic acid, butyric acid concentrations and methane production were not changed. In addition, increasing U level supplementation increased $NH_3$-N concentration. Result from real-time polymerase chain reaction revealed a significant effect on total bacteria, whereas F. succinogenes and R. flavefaciens population while R. albus was not affected by the M and U supplementation. Based on this study, it could be concluded that M and urea U supplementation could improve the nutritive value of Leucaena silage and enhance in vitro rumen fermentation efficiency. This study also suggested that the combination use of M and U supplementation level was at 2% and 1%, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.6
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• pp.843-851
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• 2003

An experiment was conducted to investigate the effects of pre-wilting Bana grass (Pennisetum purpureum x P. americanum) herbage under sunny conditions for 0, 6, 18, 24, 32 and 48 h and ensiling it with maize meal. Four levels of maize meal(viz., 0, 5, 10 and 15% on fresh weight (Fw) basis) were tested. The experiment had a split-plot design. Wilting increased the concentration of water soluble carbohydrates (WSC) significantly (p<0.001) on a Fw basis, although there were no significant changes on DM basis. Unwilted grass contained $36.1g{\cdot}WSC{\cdot}kg^{-1}{\cdot}Fw$ ($127.6g{\cdot}kg^{-1}{\cdot}DM$) and this increased to $64.1g{\cdot}WSC{\cdot}kg^{-1}{\cdot}Fw$ ($116.7g{\cdot}kg^{-1}{\cdot}DM$) after 48 h of pre-wilting. Wilting also increased the DM content of herbage significantly (p>0.001) from 250 to $620g{\cdot}kg^{-1}$, between 0 and 48 h respectively. The concentration of fermentation end-products decreased (except butyric acid) and pH increased when the period of wilting increased, indicating that fermentation was restricted. In particular, lactic acid content declined from 50.8 to $26.2g{\cdot}kg^{-1}{\cdot}DM$ (p<0.01) and the residual WSC contents of silage increased from 2.7 with fresh herbage to $18.1g{\cdot}kg^{-1}{\cdot}DM$ with 48 h of wilting (p<0.001). Rapid wilting for 24 h, to a DM of $450g{\cdot}kg^{-1}$ was optimum since important increases in pH, residual WSC and DMD occurred at this level of wilting. Acetic acid, butyric acid and ammoniacal-N contents were lowest with 24 h of wilting. There were no significant interactions between length of wilting and the incorporation of maize meal. Wilting had a greater influence on fermentation than the incorporation of maize meal. Addition of maize meal facilitated fermentation by increasing forage DM content and reducing effluent production. In addition, the maize meal increased DMD. It was concluded that maize meal should generally be incorporated at a level of 5% on fresh weight basis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.6
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• pp.529-536
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• 1986

By modified method yellow corvenia(called $Y_3$) was prepared with $4\%$ salt, $4\%$ KCl, $6\%$ sorbitol, $0.5\%$ lactic acid and $4\%$ alcohol extract of red pepper to improve the quality of fermented sea food. In this study, changes of volatile compounds and fatty acid composition obtained from modified fermented yellow corvenia($Y_3$) were experimented during fermentation, comparing with conventional fermented yellow corvenia(called $Y_1,\;20\%$ of salt contents). Total lipid of yellow corvenia was composed of $78.1\%$ of neutral lipid, $21.2\%$ of phospholipid and $0.7\%$ of glycolipid. And monoeonoic acid was held $37.4\%$ of fatty acid composition of total lipid and saturated fatty acid ($34.8\%$), polyenoic acid ($27.7\%$) were followed. Saturated fatty acid($C_{14:0},\;C_{16:0},\;C_{18:0}$) in $Y_1,\;Y_3$ increased, polyenoic acid ($C_{22:6}\;C_{22:5}\;C_{20:5}$) decreased while monoenoic acid($C_{16:1}\;C_{18:1}$) in those was little fluctuated during fermentation. Thirty-three kinds of volatile component in whole volatile compounds obtained from $Y_1,\;Y_3$ at 90 days fermentation were identified, and composed of some hydrocarbons (8 kinds), alcohols (7 kinds), acids (6 kinds), aldehydes(4 kinds), sulfides(2 kinds), ketones (2 kinds), one of phenol and 3 kinds of the other components. Among the whole volatile compounds 2-ethoxy ethanol and was held $79.35\%$ in $Y_3$, whereas nonadecane was held $75.85\%$ in $Y_1$. During fermentation 8 kinds of volatile acids, 5 kinds of amines and 9 kinds of carbonyl compounds were also detected. Those volatile acid such as acetic acid, isovaleric acid, n-caproic acid, n-butyric acid were the major portion of total volatile acids in $Y_3$ at 90 days fermentation. Meanwhile, carbonyl compounds such as ethanal, 2-butanone and butanal were the major ones, while TMA held the most part of volatile amines in $Y_3$ during fermentation. From the result of sniff test, the components which are believed to contribute to the characteristic flavor of fermented product $Y_1,\;Y_3$ are deduced to be volatile acid, carbonyl compounds and amines in order. Conclusively, there was little difference in composition of volatile components, but merely a little difference in content of those between $Y_3$ and $Y_1$.

• Asian-Australasian Journal of Animal Sciences
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• v.4 no.4
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• pp.377-382
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• 1991

The effect of molasses addition and hot temperature on the ensiling characteristics of napier grass (Pennisetum purpureum Schum.) were studied. Napier grass was harvested five times at intervals from 22 to 39 days and each harvest was divided into two equal portions. The half portion was ensiled directly and the other half was ensiled after mixing with molasses into polyethylene bag silos of 15 kg capacity. Molasses was added at the rate of 4% of fresh weight of the grass. One half of the each treatment was conserved at a room of $40^{\circ}C$ for a month and then moved to an ambient temperature room. The other half was kept at ambient temperature for the whole experimental duration. The silages were opened 3 to 7 months after ensiling. Addition of molasses enhanced lactic acid fermentation by increasing lactic acid content and reducing pH value, ammonia nitrogen and acetic, propionic and butyric acid contents of the silages in both temperature treatments. Enhanced temperature increased pH value and decreased acetic, propionic and butyric acids.

• Applied Biological Chemistry
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• v.42 no.2
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• pp.111-115
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• 1999

This study was attempted to develop technique of masking the pungent odor of chonggugjang using two-stage fermentation method with mixed pure cultures. Cooked soybeans were fermented with Lactobacilli or Aspergillus oryzae at $38^{\circ}C$ for 36 hrs, and then re-fermented with Bacillus subtilis for 12 hrs. The volatile compounds of chonggugjang were obtained with a SDE(Simultaneous steam Distillation and solvent Extraction) system and the extracts were identified by GC and GC-MS. The experimental results revealed the presence of 35 volatile compounds in control chonggugjang(only Bacillus subtilis inoculation). Among them, the major volatile compounds were 2,5-dimethyl pyrazine, 2,3,5-trimethyl pyrazine and 2-methyl pyrazine. Twenty-nine kinds of volatile compounds were in chonggugjang prepared by two-stage fermentation method with Lactobacilli and Bacillus subtilis(II), and major volatile compounds were identified to be 2,3,5-trimethyl pyrazine and 2,5-dimethyl Pyrazine. In chonggugjang(Asp. oryzae and then Bacillus subtilis inoculation(III)), the contents of hexadecanoic acid and 2-methyl pyrazine were patricularly high and the main components of chonggugjang fermented with rice straw(IV) were 2,5-dimethyl pyrazine, 2,3,5-trimethyl pyrazine etc. In conclusion, the flavor compounds such as 2, 5-dimethyl pyrazine, 2,3,5-trimethyl pyrazine and 2-methyl pyrazine were increased by the inoculation of Lactobacilli or Asp. oryzae, where as unpleasant odor components-butyric acid and valeric acid were decreased. Compared with volatile compounds of chonggugjang made from different soybean cultivars, the flavor compounds (2-methyl pyrazine 2,5-dimethyl pyrazine 2,6-dimethyl pyrazine 2,3-dimethyl pyrazine 2-acetyl pyrazine 2,3,5-trimethyl pyrazine 2-ethyl-3,5-dimethyl pyrazine) of chonggugiang prepared with Sinpaldalkong were high.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.8
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• pp.1128-1133
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• 2002

The whole plant of crop maize was chopped and ensiled in airtight 1-L capacity glass jars to determine the influence of residual sugar on anaerobic yeast growth and on the fermentation of lactic acid by L. buchneri in whole crop maize silage. There were a total of six treatments used in this experiment as follow: added 25 g de-mineralised water per kg chopped maize serving as control (con), 37.5 g glucose solution containing 12.5 g glucose ($g_1$), 75 g glucose solution containing 25 g glucose ($g_2$), 25 g L. buchneri suspension intended for $10^6$ cfu $g^{-1}$ (L.b.), $g_1+L.b.$ and $g_2+L.b.$ All silos were stored in the dark at $20^{\circ}C$ until end of experiment. Jars were opened on duplicates at day 2, 7, 14, 28, 56 or triplicates at day 91 after ensiling for measuring the pH, microbiological enumeration and fermentative products. Results indicated that acidification rates for all silages were very fast, no difference occurred among treatments before day 28. After day 28 the pH values for silages inoculated by L. buchneri. with or without sugar tended to increase especially for treated only with L. buchneri, resulting in higher (p<0.01) finial pH than uninoculated silages. Compared with control silage, the added sugar significantly (p<0.01) increased dry matter (DM) loss, L. buchneri enhanced (p<0.01) DM loss further at different sugar existence. Silages inoculated by L. buchneri only or in combination with sugar addition contained less (p<0.01) lactic acid than the correspondent silages without inoculation with L. buchneri. In comparison with control, ethanol production is about 3 or 6 fold higher due to addition 12.5 or 25 g glucose per kg chopped maize at ensiling. The added sugar resulted in less acetic acid concentration (p<0.01) than control, but inoculation with L. buchneri increased (p<0.01) acetic acid than correspondent uninoculated silages at different sugar levels. No butyric acid and propionic acid were found in uninoculted silages, silages inoculated with L. buchneri. produced more propionic acid, 1-propanol and butyric acid. Lactobacilli counts were not influenced by added sugar, but increased (p<0.01) with inoculation of L. buchneri. The added sugar increased significantly (p<0.01) the yeast count, whereas L. buchneri showed the contrary effect. No differences were found in the aerobic stability among all treatments. In conclusions, 1) the added sugars encourage the growth of yeast and yeasts convert extra sugar into ethanol in maize silages. 2) The added sugars and L. buchneri do not influence the aerobic stability of silages stored in anaerobic silos.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.1
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• pp.28-35
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• 1999

The effects of addition of celulases (A cremonium cellulolyticus and Trichoderma viride, CE), a commercial inoculum containing lactic acid bacteria (Lactobacillus casei, LAB), fermented green juice (macerated napier grass with water was incubated anaerobically with 2% glucose for 1 day, FGJ) and glucose (G), and regional difference of ensiling on napier grass (Pennisetum purpureum Schum.) silage were studied by using 900 ml laboratory glass bottle silos under 30 and $40^{\circ}C$ storage conditions in 1995 and 1996. Experiment 1 was carried out to compare the addition of CE, LAB, FGJ and the combinations. Silages were stored for 45 days after ensiling. Experiment 2 studied the effects of applications of CE, LAB, FGJ and G. Experiment 3 was carried out using the similar additives as experiment 2 except for LAB. Silages were stored for 60 days in the experiments 2 and 3. Experiments 1 and 2 were done in Nagoya, and experiment 3 in Okinawa. Sugar addition through CE or G improved the fermentation quality in all the experiments, which resulted in a greater decrease in the pH value and an increased level of lactic acid, while butyric acid contents increased under $30^{\circ}C$ storage condition in CE addition. LAB and FGJ additions hardly affected the silage fermentation quality without additional fermentable carbohydrate. But the combination of LAB, FGJ and glucidic addition (CE and G) improved the fermentation quality. The effect of the regional difference of ensiling between temperate (Nagoya; $35^{\circ}$ N) and subtropical (Okinawa; $26.5^{\circ}$ N) zones on silage fermentation quality was not shown in the present study.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.12
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• pp.1738-1742
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• 2003

The whole plant of crop maize was chopped and ensiled in double-layered polyethylene bags to determine the influence of residual sugar on the fermentation of lactic acid and aerobic stability by L. buchneri in whole crop maize silage made in airstress condition. There were a total of six treatments used in this experiment as follow: added 25 g de-mineralised water per kg chopped maize serving as control (con), 37.5 g glucose solution containing 12.5 g glucose ($g_1$), 75 g glucose solution containing 25 g glucose ($g_2$), 25 g, L,.buchneri suspension intended for $10^6$ cfu $g^{-1}$ (L.b.), $g_1$+L.b. and $g_2$+L.b. All silos were opened at day 91 after ensiling for measuring the pH values, microbiological enumeration, fermentative products and aerobic stability. The dry matter loss increased significantly (p<0.01) due to inclusion of sugar or L. buchneri. The lower lactic acid concentrations were observed (p<0.01) in silages inoculated with L. buchneri only or in combination with sugar addition than the correspondent uninoculated silages. Compared with control silage, ethanol production was about 3 or 6-fold higher due to addition 12.5 or 25 g glucose per kg chopped maize at ensiling. The silages added with sugar contained less acetic acid concentration (p<0.01) than control, but silages inoculated with L. buchneri showed the contrary effects (p<0.01) at different sugar levels. No butyric acid was found in uninoculated silages, silages inoculated with L. buchneri. producted more propionic acid, 1-propanol and butyric acid. Lactic acid bacteria counts increased markedly (p<0.01) due to inoculation with L. buchneri, whereas it was reduced (p<0.01) by added sugar. No significant difference was observed in count of yeast, but inoculation with L. buchneri shows a decreasing trend. Mould count in all silages was less than 2 (log cfu $g^{-1}$). The added sugar had negative effects on aerobic stability of maize silage made under air-stress conditions, whereas inoculation with L. buchneri improves (p<0.01) the aerobic stability.