• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.1
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• pp.1-7
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• 1997

In order to utilize fish by-products from the skipjack tuna (Katsuwonus pelamis) canning manufactures Lactobacillus buigaricus KCTC 3188 and L. piantarum KCTC 1048 were used as a starter culture for the preparation of fermented fish silage with skipjark tuna viscera. The optimum temperature and pH on barterial growth and lactic acid production of L. bulgaricus and L. plantarum in MRS broth were $35^{\circ}C$ and around pH 6.0, respectively. And the optimum concentrations of the carbohydrate sources added to the broths were $7\%$ for dextrose and $10\%$ for molasses on the basis of total weights of skipjack tuna viscera. The pH of acid treated skipjack tuna viscera silage (ASS) slightly increased from 4.0 to 4.5, while that of fermented skipjack tuna viscera silages by the use of lactic acid bacterias (FSS) was significantly declined from 5.9 to about 40 after 42 days of storage at $35^{\circ}C$. Though the content of volatile basie nitrogen (VBN) in ASS was lower than those of FSS after 42 days of storage at $35^{\circ}C$, VBN content in silages slightly increased from an initial value of $62\~65{\cdot}mg/100g$ to final value of $113\~155\;mg/100g$ over 42 days. The fermented silage by L. piantarum reached a maximum concentration of amino nitrogen and showed $81\%$ of hydrolysis degree after 4 days of storage at $35^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.10
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• pp.1528-1539
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• 2019

Objective: To evaluate the effects on microbial diversity and biochemical parameters of gradually increasing temperatures, from $5^{\circ}C$ to $25^{\circ}C$ on corn silage which was previously fermented at ambient or low temperature. Methods: Whole-plant corn silage was fermented in vacuum bag mini-silos at either $10^{\circ}C$ or $20^{\circ}C$ for two months and stored at $5^{\circ}C$ for two months. The mini-silos were then subjected to additional incubation from $5^{\circ}C$ to $25^{\circ}C$ in $5^{\circ}C$ increments. Bacterial and fungal diversity was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) profiling and biochemical analysis from mini-silos collected at each temperature. Results: A temperature of $10^{\circ}C$ during fermentation restricted silage fermentation compared to fermentation temperature of $20^{\circ}C$. As storage temperature increased from $5^{\circ}C$ to $25^{\circ}C$, little changes occurred in silages fermented at $20^{\circ}C$, in terms of most biochemical parameters as well as bacterial and fungal populations. However, a high number of enterobacteria and yeasts (4 to $5\;log_{10}$ colony forming unit/g fresh materials) were detected at $15^{\circ}C$ and above. PCR-DGGE profile showed that Candida humilis predominated the fungi flora. For silage fermented at $10^{\circ}C$, no significant changes were observed in most silage characteristics when temperature was increased from $5^{\circ}C$ to $20^{\circ}C$. However, above $20^{\circ}C$, silage fermentation resumed as observed from the significantly increased number of lactic acid bacteria colonies, acetic acid content, and the rapid decline in pH and water-soluble carbohydrates concentration. DGGE results showed that Lactobacillus buchneri started to dominate the bacterial flora as temperature increased from $20^{\circ}C$ to $25^{\circ}C$. Conclusion: Temperature during fermentation as well as temperature during storage modulates microorganism population development and fermentation patterns. Silage fermented at $20^{\circ}C$ indicated that these silages should have lower aerobic stability at opening because of better survival of yeasts and enterobacteria.

• Animal Bioscience
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• v.35 no.9
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• pp.1379-1389
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• 2022

Objective: This study identified the major lactic acid bacteria (LAB) strains from different fermented total mixed rations (FTMRs) via metataxonomic analysis and evaluated the ability of their standard strain as ensiling inoculants for corn stover silage. Methods: The bacterial composition of eight FTMRs were analyzed by 16S rDNA sequencing. Corn stover was ensiled without LAB inoculation (control) or with 1×10⁶ cfu/g LAB standard strain (Lactobacillus vaginalis, Lactobacillus reuteri, Lactobacillus helveticus, or Lactobacillus paralimentarius) selected from the FTMRs or 10 g/t commercial silage inoculant (CSI) around 25℃ for 56 days. For each inoculation, a portion of the silage was sampled to analyze ensiling characteristics at time intervals of 0, 1, 3, 7, 14, 28, and 56 days, gas production (GP), microbial crude protein and volatile fatty acids as the measurements of rumen fermentation characteristics were evaluated in vitro with the silages of 56 days after 72 h incubation. Results: Lactobacillus covered >85% relative abundance of all FTMRs, in which L. pontis, L. vaginalis, L. reuteri, L. helveticus, and L. paralimentarius showed >4% in specific FTMRs. CSI, L. helveticus, and L. paralimentarius accelerated the decline of silage pH. Silage inoculated with L. paralimentarius and CSI produced more lactic acid the early 14 days. Silage inoculated with L. paralimentarius produced less acetic acid and butyric acid. For the in vitro rumen fermentation, silage inoculated with CSI produced more potential GP, isobutyric acid, and isovaleric acid; silage inoculated with L. helveticus produced more potential GP and isovaleric acid, silage inoculated with L. paralimentarius or L. reuteri produced more potential GP only. Conclusion: The standard strain L. paralimentarius (DSM 13238) is a promising ensiling inoculant for corn stover silage. The findings provide clues on strategies to select LAB to improve the quality of silage.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.6
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• pp.807-811
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• 2005

This experiment examined the characteristics of fermented juice of epiphytic lactic acid bacteria (FJLB) prepared by the addition of glucose, sucrose and molasses as a fermentation substrate. The effect of FJLB on the fermentative quality and changes in chemical composition during fermentation of guineagrass silage were also investigated. The pH value of the silages treated with FJLB rapidly decreased, and reached to the lowest value within 7 days of start of fermentation, as compared to the control. The number of lactic acid bacteria (LAB) in the treated silages increased for the first 3 days, thereafter the number of LAB declined gradually up to the end of the experiment. Silages treated with FJLB had larger populations of LAB than the control. Ammonia-nitrogen production increased throughout the ensiling period, which in the control and no-sugar added FJLB silages were higher than the other treated silages. Lactic acid levels varied with the time of ensiling and among the silage treatments. For any sugar FJLB treated silages, the lactic acid increased initially, and then slightly reduced to less than 50 g/kg of dry matter until 49 days after ensiling, except the silage treated with glucose added FJLB. Nevertheless, lactic acid content of the control decreased constantly from the beginning of ensiling and was not found after 35 days. Moreover, acetic acid content increased throughout the ensiling period. All the FJLB treated silages had significantly (p<0.05) lower pH and ammonia-nitrogen content, while significantly (p<0.05) higher lactic acid content and V-score value compared with the control. This study confirmed that the applying of FJLB with any sugar substrate improved fermentative quality of silage.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.7
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• pp.942-946
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• 2004

This study aimed to examine the fermentation quality of a tropical Elephant grass (Pennisetum purpuereum) and temperate Italian ryegrass (Lolium multiflorum) forages ensiled additive of fermented juice of epiphytic lactic acid bacteria (LAB) and to determine what factor affects the fermentation characteristics of the crops. In both species cell walls neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents in silages were consistently decreased (p<0.05) with the addition of FJLB at ensiling more then Control treatment. The FJLB additive increased number of LAB (cfu) and lactate concentration in the silages in both species. The Control treatment without additive underwent a clostridial type of fermentation with traces of propionic, iso-butyric, n-butyric acids contents with higher (p<0.01) levels of volatile basic nitrogen (VBN %TN) and had appreciable decreased of nutrient in silages. FJLB treatment improved silage nutritive value with little contents of VBN %TN, ethanol and very small amount of dry matter (DM) and hemicellulose losses (p<0.05) between 2 to 5% and 7 to 3% respectively, in Elephant grass and Italian ryegrass species. The results in this study indicates that while among the factors affecting silage fermentation butyric type of fermentation was more pronounced in tropical elephant grass compared to the temperate Italian ryegrass, FJLB additive revealed a better silage fermentation products in both species.

• Journal of Animal Science and Technology
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• v.44 no.6
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• pp.711-718
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• 2002

In order to investigate the effect of feeding browses on the incidence of abortion in pregnant Korean native female goats, 20 pregnant does(3${\sim}$4 month) were used as experimental animal and they grouped 4 treatments with 5 heads according to experimental diets. Each group was fed pine browse, pine browse silage, fermented pine browse, or oak browse silage. Browses intake a day was 0.36kg for oak browse silage, 0.28kg for pine browse, 0.24kg for pine browse silage, and 0.14kg for fermented pine browse. Salmonella and Fungi were not found in pine browses but they were found in oak browse silage in the amount of 7.93${\times}$$10^3$ and 11.1${\times}$$10^1$ cfu/g, respectively. E. Coliwas found 11.67${\times}$$10^1$cfu/g in oak browse silage. The incidence of abortion was 60% for fermented pine browse, 40% for pine browse silage, and 20% for pine browse feeding. Abortion did not occur by feeding oak browse silage. Progesterone concentration was similar each other regardless of normal delivery or abortion at delivery day but the concentration of estradiol was higher for normal delivery, concentration of cortisol was decreased until the delivery day in normal but increased in abortion does. The results were suggested.

• Animal Bioscience
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• v.36 no.5
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• pp.720-730
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• 2023

Objective: This study investigated the effects of corn silage as a source of microbial inoculant containing antifungal and carboxylesterase-producing bacteria on fermentation, aerobic stability, and nutrient digestibility of fermented total mixed ration (FTMR) with different energy levels. Methods: Corn silage was used as a bacterial source by ensiling for 72 d with an inoculant mixture of Lactobacillus brevis 5M2 and L. buchneri 6M1 at a 1:1 ratio. The corn silage without or with inoculant (CON vs MIX) was mixed with the other ingredients to formulate for low and high energy diets (LOW vs HIGH) for Hanwoo steers. All diets were ensiled into 20 L mini silo (5 kg) for 40 d in quadruplicate. Results: The MIX diets had lower (p<0.05) acid detergent fiber with higher (p<0.05) in vitro digestibilities of dry matter and neutral detergent fiber compared to the CON diets. In terms of fermentation characteristics, the MIX diets had higher (p<0.05) acetate than the CON diets. The MIX diets had extended (p<0.05) lactic acid bacteria growth at 4 to 7 d of aerobic exposure and showed lower (p<0.05) yeast growth at 7 d of aerobic exposure than the CON diets. In terms of rumen fermentation, the MIX diets had higher (p<0.05) total fermentable fraction and total volatile fatty acid, with lower (p<0.05) pH than those of CON diets. The interaction (p = 0.036) between inoculant and diet level was only found in the immediately fermentable fraction, which inoculant was only effective on LOW diets. Conclusion: Application of corn silage with inoculant on FTMR presented an antifungal effect by inhibiting yeast at aerobic exposure and a carboxylesterase effect by improving nutrient digestibility. It also indicated that fermented feedstuffs could be used as microbial source for FTMR. Generally, the interaction between inoculant and diet level had less effect on this FTMR study.

• 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.5 no.2
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• pp.249-256
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• 1992

Italian ryegrass (Lolium multiflorum Lam.) was fractionated into leaf nutrient concentrate, fiber and deproteinised juice (DPJ). The fresh and fermented DPJs were concentrated and referred to as fresh deproteinised juice concentrate (FDPJC) and fermented deprotenised juice concentrate (FMTD DPJC). The FDPJC and FMTD DPJC were separately mixed with dried fiber and ensiled. Wilted crop silage and fresh fiber silage were also prepared from the same material crop. The nutritive value of these four silages were compared using four goats by $4{\times}4$ Latin square design. Green crop fractionation resulted lesser amount of crude protein and ash, and higher amount of neurtal detergent fiber, acid detergent fiber, cellulose and hemicellulose in fresh fiber. The pH of fresh fiber silage was lower than that of the other silages. Addition of FDPJC or FMTD DPJC to the dried fiber at ensiling did not improve the silage qualities; but all the silages were satisfactorily preserved. Goats fed these silages showed similar ruminal pH and total volatile fatty acid concentrations. But addition of FMTD DPJC was effective on increasing ruminal acetic acid concentration reducing propionic acid concentration. Ruminal n- and iso- butyric acid concentrations were proportional to that of propionic acid. High ammonia content of the silage containing FMTD DPJC was reflected to the ruminal ammonia concentration, urinary nitrogen excretion and serum urea level of goats. Inclusion of FDPJC or FMTD DPJC added 15 to 25% dry matter to the fiber silages with a little reduction in the digestibilities of most components of the silages.

• Animal Bioscience
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• v.34 no.4
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• pp.642-651
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• 2021

Objective: This study aimed to determine the effects of different roughages in total mixed ration (TMR) inoculated with or without coculture of Lactobacillus acidophilus (L. acidophilus) and Bacillus subtilis (B. subtilis) on in vitro rumen fermentation and microbial population. Methods: Three TMRs formulations composed of different forages were used and each TMR was grouped into two treatments: non-fermented TMR and fermented TMR (F-TMR) (inoculated with coculture of L. acidophilus and B. subtilis). After fermentation, the fermentation, chemical and microbial profile of the TMRs were determined. The treatments were used for in vitro rumen fermentation to determine total gas production, pH, ammonianitrogen (NH3-N), and volatile fatty acids (VFA). Microbial populations were determined by quantitative real-time polymerase chain reaction (PCR). All data were analyzed as a 3×2 factorial arrangement design using the MIXED procedure of Statistical Analysis Systems. Results: Changes in the fermentation (pH, lactate, acetate, propionate, and NH3-N) and chemical composition (moisture, crude protein, crude fiber, and ash) were observed. For in vitro rumen fermentation, lower rumen pH, higher acetate, propionate, and total VFA content were observed in the F-TMR group after 24 h incubation (p<0.05). F-TMR group had higher acetate concentration compared with the non-fermented group. Total VFA was highest (p<0.05) in F-TMR containing combined forage of domestic and imported source (F-CF) and F-TMR containing Italian ryegrass silage and corn silage (F-IRS-CS) than that of TMR diet containing oat, timothy, and alfalfa hay. The microbial population was not affected by the different TMR diets. Conclusion: The use of Italian ryegrass silage and corn silage, as well as the inoculation of coculture of L. acidophilus and B. subtilis, in the TMR caused changes in the pH, lactate and acetate concentrations, and chemical composition of experimental diets. In addition, F-TMR composed with Italian ryegrass silage and corn silage altered ruminal pH and VFA concentrations during in vitro rumen fermentation experiment.