Dynamics of Early Fermentation of Italian Ryegrass (Lolium multiflorum Lam.)Silage

Shao, Tao,Ohba, N.,Shimojo, M.,Masuda, Y. Asian Australasian Association of Animal Productio 2002 Animal Bioscience Vol.15 No.11

The dynamics of fermentation were studied with Italian ryegrass ensiled in the laboratory silos. The silos were kept in the room set at 25$^{\circ}C$, and then were opened on 0.5, 1, 2, 3, 5, 7 and 14 days after ensiling, respectively. The samples were taken from three silos at each sampling time for chemical analyses. Mono-and disaccharides composition was determined for glucose, fructose and sucrose by high performance liquid chromatography. The Italian ryegrass silage succeeded to achieve lactate type fermentation; high values of lactic acid (85.83 g/kg) and lactic acid/acetic acid at the end of ensiling (14 day), low values of pH (3.74), acetic acid (5.38 g/kg), ethanol (19.20 g/kg) and $NH_3-N/Total\;N$ (75.84 g/kg), no or only small amounts of butyric acid, valeric acid and propionic acid. The fermentation dynamics showed a fast and large pH decrease caused by a fast and large production of lactic acid during the first 5 days. Mono-and disaccharides composition largely decreased within initial 0.5 day (12 h) of ensiling. Sucrose disappeared rapidly within initial 0.5 day of ensiling, and fructose and glucose contents showed an initial rise during ensiling, and then decreased gradually. These indicated that the enzymes of plant tissue were active within 2 days of ensiling, which caused the initial rise in fructose and glucose from the hydrolysis of sucrose and fructans. After 5 days of ensilage, glucose was consumed completely, suggesting that glucose was the first fermentation substrate. After 2 days of ensiling, sum amounts of lactic acid and remaining mono-and disaccharides proved to be larger than the quantity of mono-and disaccharides in the initial grass. From the facts mentioned above, it was suggested that considerable amounts of lactic acid were produced from some other substrate such as fructans than initial mono-and disaccharides.

Delta-endotoxin proteins associated with spherical parasporal inclusions of the four Lepidoptera-specific Bacillus thuringiensis strains

Kim, K.-H.,Wasano, N.,Ohba, M. 동의대학교 기초과학연구소 1999 基礎科學硏究論文集 Vol.9 No.1

Four Lepidoptera-specific reference strains of Bacillus thuringiensis, belonging to serovars sumiyoshiensis(H3a:3d), fukuokaensis(H3a:3d:3e), darmstadiensis(H10a:10b) and japonensis(H23), which produce spherical parasporal inclusions, were examined for comparative characterization of δ-endotoxins. SDS-PAGE profiles of the alkali-solubilized parasporal inclusions revealed the presence of single major protein bands of 130 kDa in the four strains. Chymotrypsin and trypsin treatment of the proteins gave profiles different from those of the strains HD-1(serovar kurstaki, H3a:3b:3c) and T84Al(serovar sotto, H4a:4b). Also, minor variations were observed in proteolysis profiles among the four strains. The LC_50 values of purified parasporal inclusions for the silkworm(Bombyx mori) larvae were 7·35, 6·45, 3·08 and 2·63 ㎍ g^-1 diet, respectively, showing that their toxicity levels were 5-15 times lower than that of the strain HD-1 (0·49㎍ g^-1 diet). Analysis by immunodiffusion and immunoblotting with polyclonal antisera revealed that parasporal inclusion proteins of the four strains are highly related, whereas they shared few or no common anitgens with those of the strains HD-1, T84Al and Buibui(serovar japonensis).

Effects of Preheated Additives on the Fermentation Quality of Napiergrass Silage

Yunus, M.,Ohba, N.,Tobisa, M.,Shimojo, M.,Masuda, Y. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.11

Hydrolysis of plant protein to non-protein nitrogen (N) or ammonia can reduce quality of silage crops. Heating or non-enzymatic browning is a treatment to inhibit this hydrolysis. This experiment was conducted to examine the effects of pre-heated soybean meal and molasses on the fermentation quality of napiergrass silage. The initial growth of napiergrass was harvested at 85 days of age and immediately chopped into about 1 cm length. About 700 g of the grass was ensiled into a laboratory silo (1.0 liter polyethylene container) and incubated for 30 days at room temperature ($28^{\circ}C$). No additives (control), molasses, soybean meal and molasses + soybean meal treatments were prepared. All additives were non-heated or heated in an oven at $150^{\circ}C$ for 30 minutes before ensiling. Molasses was added at 3% on the fresh weight basis and soybean meal was added at 0.5% N, respectively. After opening the silo, pH, total nitrogen (TN), volatile basic nitrogen (VBN), lactic acid (LA), acetic acid (AA), butyric acid (BA) and dry matter (DM) contents were determined. The data were analyzed statistically by analysis of variance. Compared with control, molasses addition significantly decreased pH value, VBN/TN, AA and BA and increased LA production. Soybean meal addition significantly increased TN and VBN/TN of silage. Both molasses and soybean meal addition significantly reduced pH value, AA, and BA and increased DM and LA contents of silage. The heating of additives was only effective to reduce VBN/TN production compared with non-heated additives in soybean meal and soybean meal with molasses addition.

Effect of Glucose and Formic Acid on the Quality of Napiergrass Silage After Treatment With Urea

Yunus, M.,Ohba, N.,Tobisa, M.,Shimojo, M.,Masuda, Y. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.2

Urea as a silage additive increases crude protein but reduces fermentation quality of silage by increasing pH and enhancing clostridial bacteria growth, especially in low sugar forages. Glucose and formic acid might be expected to compensate these defects caused by urea addition to grass silage. Thus, in this experiment urea formic acid or urea with glucose was applied to improve N content and the quality of napiergrass (Pennisetum purpureum Schumach.) silage. The first growth of napiergrass was harvested at 85 days of age and about 700 g of the grass was ensiled in laboratory silos (1.0 liter polyethylene containers) for 2, 7, 14, and 30 days at room temperature ($28^{\circ}C$). The treatments were no additives (control), urea, urea+glucose or urea+formic acid. Urea was added before ensiling at 0.5% of fresh weight of napiergrass and glucose and formic acid were added at 1% of fresh weight, respectively. After opening the silo, pH, dry matter content (DM), contents on DM basis of total N (TN), volatile basic nitrogen (VBN), lactic acid (LA), acetic acid (AA) and butyric acid (BA) were determined. The control at 30 days of fermentation showed 5.89 for pH with 13.8% for VBN/TN and 1.51% for AA. The addition of urea increased TN by about 1.5% units but decreased the fermentation quality by increasing pH from 5.89 to 6.86, increasing VBN/TN from 13.8% to 24.63%, increasing BA from 0.02% to 0.56%, and decreasing LA from 1.03% to 0.02%. Glucose addition with urea significantly decreased VBN/TN from 13.8% to 4.44% by reducing pH from 6.86 to 4.83 because of higher production of LA (2.62%). Adding urea and formic acid resulted in a more pronounced depression of VBN/TN and fermentation than the addition of urea and glucose. This study suggested that the combination of 1% glucose or 1% formic acid with 0.5% urea will improve nutritive value and fermentation quality of napiergrass silage.

Effects of Adding Glucose, Sorbic Acid and Pre-fermented Juices on the Fermentation Quality of Guineagrass (Panicum maximum Jacq.) Silages

Shao, Tao,Ohba, N.,Shimojo, M.,Masuda, Y. Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.6

This study was conducted to evaluate the effects of adding glucose (G), sorbic acid (S), pre-fermented juice of epiphytic lactic acid bacteria (FJLB) and their combinations on the fermentation qualities and residual mono-and di-saccharides compositions of guineagrass silage. The additives used in this experiment were 1% glucose, 0.1% sorbic acid and FJLB at a theoretical application rate of 9.0${\times}$105 CFU $g^{-1}$ on the fresh weight basis of guineagrass, respectively. There was a total of eight treatments in this experiment: (1) C (without additives), (2) FJLB, (3) S, (4) G, (5) FJLB+S, (6) FJLB+G, (7) S+G, (8) FJLB+S+G. After 30 days of storage, the silos were opened for chemical analyses. Based on the results, all additives were efficient in improving the fermentation quality of guineagrass silage. This was well indicated by significantly (p<0.05) lower pH and BA content and significantly (p<0.05) higher LA content in the treated silages except for the FJLB than in the C. However, there was only a slight increase in LA for the FJLB as compared with the C, which might be due to the low WSC content of the original guineagrass (34.4 g $kg^{-1}$). When the FJLB+S and FJLB+G were added, there were significant (p<0.05) decreases in pH and significant (p<0.05) increases in LA as compared with the FJLB alone. This indicated that the G, S and FJLB were of synergestic effects on the silage fermentation quality. The G combination treatments including the G alone showed large improvements in the fermentation quality as compared with the treatments without the G. This suggested that adding fermentable substrates (G) to plant materials such as guineagrass, which contain low WSC, intermediate population of epiphytic LAB, CP and DM content, is more important and efficient for improving the fermentation quality of silages than adding a number of species of domestic LAB (FJLB) and aerobic bacteria inhibitor (S).

Improving Fermentation and Nutritive Quality of Napiergrass Silage by Mixing with Phasey Bean

Yunus, M.,Ohba, N.,Tobisa, M.,Nakano, Y.,Shimojo, M.,Furuse, M.,Masuda, Y. Asian Australasian Association of Animal Productio 2001 Animal Bioscience Vol.14 No.7

To improve the low content of crude protein and low quality fermentation of napiergrass silage, ensiling with phasey bean was examined. Napiergrasses, (Pennisetum purpureum cv. Merkeron), hybrid napiergrass (Pennisetum purpureum${\times}$ Pennisetum typhoides) and phasey bean (Macroptilium lathyroides cv. Murray) were cultivated in Fukuoka, Japan. The first growth of napiergrasses and phasey bean were harvested at 90 days of age and chopped into about 1 cm lengths. The mixing levels of phasey bean with napiergrasses were 0, 25, 50 and 75; 700 g of plant materials were ensiled into a laboratory silo (1.0 litre. polyethylene container) and incubated for 30 days. After opening silos, pH, total nitrogen (TN), volatile basic nitrogen (VBN) and organic acids (lactic acid: LA, acetic acid: AA, butyric acid: BA) were determined. The experiment was a factorial design of $2{\times}4$ with 3 replicates. The data were analyzed statistically by two-way analysis of variance. Without inclusion of phasey bean, lower quality was observed for hybrid silage than for Merkeron silage. The inclusion of phasey bean at the rate of 25% increased DM and TN, and decreased pH value, VBN/TN, AA and BA in both Merkeron and hybrid napiergrass silages. The value of pH, TN, VBN/TN, AA and BA were not significantly different among 25, 50 and 75% mixing levels of phasey bean. LA production increased significantly with the increase in phasey bean mixing level in both hybrid and Merkeron silages. At all mixing levels of phasey bean, LA production did not show significant differences between Merkeron and hybrid silages.

Effects of Adding Urea and Molasses on Napiergrass Silage Quality

Yunus, M.,Ohba, N.,Shimojo, M.,Furuse, M.,Masuda, Y. Asian Australasian Association of Animal Productio 2000 Animal Bioscience Vol.13 No.11

To standardize proper formulation of urea and molasses, the former to increase crude protein content of tropical grass and the latter for improving its silage quality, we examined the fermentation quality of silage of fresh and wilted napiergrass (Pennisetum purpureum Schumach) with different levels of urea and molasses with or without lactic acid bacteria (LAB). Silage was made of napiergrass with conditions of fresh young (Exp. 1),young wilted for half day (Exp. 2) and fresh mature (Exp. 3). Chopped plant materials of about 1cm length were ensiled into a laboratory silo and incubated for one month at $25^{\circ}C$. The treatments were the combination of 0, 0.2 and 0.6% of urea and 0, 2 and 5% of molasses (fresh material basis) with or without LAB inoculation. After opening the silo, pH, organic acids, volatile basic nitrogen (VBN) and total nitrogen (TN) were determined. Addition of molasses significantly (p<0.01) lowered pH values in three experiments. Though molasses addition increased lactic acid production even at a higher level of urea, pH values at 0 and 2% molasses were significantly increased by urea in fresh and wilted young silages, but in fresh mature silage it occurred only when molasses was not added. VBN/TN at 0.6% urea were decreased significantly by the highest molasses in three experiments. Significant increases in TN by the increasing of urea addition were observed at all levels of molasses in wilted young and fresh mature silages. In conclusion, a combination of 5% molasses and 0.6% urea could improve the nutritive and fermentation qualities of napiergrass silage under young, wilting and mature conditions.

Sexual Dimorphism in the Compound Eye of Rhagophthalmus ohbai (Coleoptera: Rhagophthalmidae): II. Physiology and Function of the Eye of the Male

T.F.S. Lau,N. Ohba,K. Arikawa,V.B. Meyer-Rochow 한국응용곤충학회 2007 Journal of Asia-Pacific Entomology Vol.10 No.1

The eyes of male and female Rhagophthalmus ohbai are of very different sizes and possess approximately 3000 and 35 facets, respectively. In the male eye one can distinguish a smaller dorsal region with 500 facets and a larger ventral one with ca. 1800. Ultrastructural differences between them have been described earlier in this journal (Lau and Meyer- Rochow, 2006). Electrophysiological recordings from the two eye areas have now revealed that the ventral region is maximally sensitive to light of 600 nm wavelength, while the dorsal eye region responds maximally to light of 540-560 nm wavelengths. In the dorsal eye region sensitivity to UV-radiation at around 360 nm wavelength, being twice as high as that of the ventral eye region, amounted to ca. one quarter of peak wavelength sensitivity. The regional differences in spectral sensitivity seem to be a reflection of the different tasks of the two eye regions: looking downward to see the yellow light emitted by a female, sensitivity towards longer wavelengths would be advantageous, but looking upward into the twilight sky, sensitivity to shorter wavelength would be a more appropriate adaptation.

Sexual Dimorphism in the Compound Eye of Rhagophthalmus ohbai (Coleoptera: Rhagophthalmidae): II. Physiology and Function of the Eye of the Male

Lau, T.F.S.,Ohba, N.,Arikawa, K.,Meyer-Rochow, V.B. Korean Society of Applied Entomology 2007 Journal of Asia-Pacific Entomology Vol.10 No.1

The eyes of male and female Rhagophthalmus ohbai are of very different sizes and possess approximately 3000 and 35 facets, respectively. In the male eye one can distinguish a smaller dorsal region with 500 facets and a larger ventral one with ca. 1800. Ultrastructural differences between them have been described earlier in this journal (Lau and Meyer-Rochow, 2006). Electrophysiological recordings from the two eye areas have now revealed that the ventral region is maximally sensitive to light of 600 nm wavelength, while the dorsal eye region responds maximally to light of 540-560 nm wavelengths. In the dorsal eye region sensitivity to UV-radiation at around 360 nm wavelength, being twice as high as that of the ventral eye region, amounted to ca. one quarter of peak wavelength sensitivity. The regional differences in spectral sensitivity seem to be a reflection of the different tasks of the two eye regions: looking downward to see the yellow light emitted by a female, sensitivity towards longer wavelengths would be advantageous, but looking upward into the twilight sky, sensitivity to shorter wavelength would be a more appropriate adaptation.

Realtime Measurement of Impedance Locus using Impedance Spectroscopy : How Many and How Low Frequencies Are Required ?

T. Fukumoto,G. M. Eom,S. Ohba,N. Hoshimiya 한국정밀공학회 2004 한국정밀공학회 학술발표대회 논문집 Vol.2004 No.10월

High temporal-resolution and accurate measurement of skin impedance locus provides useful data for the identification of the physiological/psychological changes and also the identification of acupuncture point. An impedance spectroscopy method using digitally constructed current waveform consisting of many frequency components (multiples of 1Hz) was reported3. The time resolution of the method depends on the lowest frequency used in the waveform construction, and therefore, the measurement would be faster if the lowest frequency is the higher. However, it was not clear that how many and how low frequencies must be used for the estimation of the skin impedance parameters from which the impedance locus can be drawn. This study shows the relationship between the estimation error of the impedance parameters and the frequency coverage of the spectroscopy. The results of this study are expected to serve as the reference of the frequency selection in the impedance spectroscopy.