• Title/Summary/Keyword: In vitro degradation

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Effects of Strains of Saccharomyces cerevisiae and Incubation Conditions on the In vitro Degradability of Yeast and Roughage

  • Ando, S.;Nishiguchi, Y.;Hayasaka, K.;Yoshihara, Y.;Takahashi, J.;Iefuji, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.3
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    • pp.354-357
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    • 2005
  • The in vitro degradability of yeast and the effect of yeast on the in vitro degradability of forage may differ in terms of the specific yeast strains or their incubation conditions. Thus in experiment 1, two strains of sake yeast (strainK7 and strainK9) and one strain of bakers' yeast (KY5649) were incubated in an aerobic condition. In experiment 2, aerobically or anaero bically incubated K7 was used for investigating the in vitro degradability of yeast, the effect of yeast on the in vitro degradability of forage, and the degradability of yeast by pepsin and pronase treatment. The in vitrodegradability of bakers' yeast was significantly (p<0.05) higher than those of sake yeasts. The in vitro degradability of anaerobically incubated yeast was significantly (p<0.01) higher than that of aerobically incubated yeast. The degradability of bakers' yeast by pepsin treatment was significantly (p<0.01) higher than that of the sake yeasts. The degradability of bakers' yeast by pronase treatment was slightly higher than that of the two sake yeasts, while the degradability of anaerobically incubated yeast by both enzymes, respectively, was significantly (p<0.01) higher than that of aerobically incubated yeast. The degradability of forages was increased significantly (p<0.05) by the addition of yeasts. The degradability of roughage by sake yeast tended to be higher than that by the bakers' yeast. The degradability of roughage was significantly (p<0.05) higher by anaerobically incubated yeast than by aerobically incubated yeast. Given the above results, it seems that in vitro degradability of yeast and the magnitude of the increment of roughage degradation differ among the yeast strains and their incubation conditions.

Effects of Defaunation on Fermentation Characteristics and Methane Production by Rumen Microbes In vitro When Incubated with Starchy Feed Sources

  • Qin, W.Z.;Li, C.Y.;Kim, J.K.;Ju, J.G.;Song, Man-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.10
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    • pp.1381-1388
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    • 2012
  • An in vitro experiment was conducted to examine the effects of defaunation (removal of protozoa) on ruminal fermentation characteristics, $CH_4$ production and degradation by rumen microbes when incubated with cereal grains (corn, wheat and rye). Sodium lauryl sulfate as a defaunation reagent was added into the culture solution at a concentration of 0.000375 g/ml, and incubated anaerobically for up to 12 h at $39^{\circ}C$. Following defaunation, live protozoa in the culture solution were rarely observed by microscopic examination. A difference in pH was found among grains regardless of defaunation at all incubation times (p<0.01 to 0.001). Defaunation significantly decreased pH at 12 h (p<0.05) when rumen fluid was incubated with grains. Ammonia-N concentration was increased by defaunation for all grains at 6 h (p<0.05) and 12 h (p<0.05) incubation times. Total VFA concentration was increased by defaunation at 6 h (p<0.05) and 12 h (p<0.01) for all grains. Meanwhile, defaunation decreased acetate and butyrate proportions at 6 h (p<0.05, p<0.01) and 12 h (p<0.01, p<0.001), but increased the propionate proportion at 3 h, 6 h and 12 h incubation (p<0.01 to 0.001) for all grains. Defaunation increased in vitro effective degradability of DM (p<0.05). Production of total gas and $CO_2$ was decreased by defaunation for all grains at 1 h (p<0.05, p<0.05) and then increased at 6 h (p<0.05, p<0.05) and 12 h (p<0.05, p<0.05). $CH_4$ production was higher from faunation than from defaunation at all incubation times (p<0.05).

A COMPARATIVE STUDY OF THE PROTECTION OF DHAINCHA (Sesbania aculeata) SEED MEAL AND FISH MEAL FROM RUMEN DEGRADATION USING NYLON BAG TECHNIQUE

  • Hussain, M.;Chowdhury, B.;Siddiqua, A.;Routh, C.K.;Saadullah, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.2
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    • pp.197-203
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    • 1993
  • The protection of dry matter and nitrogen from rumen degradation of dhaincha seed (Sesbania aculeate) meal was studied using nylon bag technique. The dhaincha seed meal was subjected to various heat treatments that included oven drying, autoclaving and boiling. Similar experiment was conducted with fish meal as reference for comparison. The oven-dried dhaincha meal was found to retain more dry matter and nitrogen than was found boiled or autoclaved meal. While autoclaving appeared to improve nitrogen and dry matter retention to some extent, boiling seemed to cause more loss of dry matter from nylon bag. Heat treatment caused high retention of nitrogen by fish meal. The calculated effective protein degradation was 80.4% and 83.2% for the oven dried fish meal and dhaincha seed meal whereas same values were 74.2% and 86.7% for autoclaved fish and dhaincha seed meal respectively at the outflow rate of 4.4% per hour. The in vitro study revealed higher digestibility for heat treated samples by pepsin. The dry matter, nitrogen and ash content of dhaincha seed meal were 85.93%, 5.93% and 7.31% respectively.

Enhanced In Vitro Protein Synthesis Through Optimal Design of PCR Primers

  • Ahn Jin-Ho;Son Jeong-Mi;Hwang Mi-Yeon;Kim Tae-Wan;Park Chang-Kil;Choi Cha-Yong;Kim Dong-Myung
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.355-359
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    • 2006
  • The functional stability of mRNA is one of the crucial factors affecting the efficiency of in vitro translation. As the rapid degradation of mRNA in the cell extract (S30 extract) causes early termination of the translational reactions, extending the mRNA half-life will improve the productivity of the in vitro protein synthesis. Thus, a simple PCR-based method is introduced to increase the stability of mRNA in an S30 extract. The target genes are PCR-amplified with primers designed to make the ends of the transcribed mRNA molecule anneal to each other. When compared with normal mRNA, the mRNA with the annealing sequences resulted in an approximately 2-fold increase of protein synthesis in an in vitro translation reaction. In addition, sequential transcription and translation reactions in a single tube enabled direct protein expression from the PCR-amplified genes without any separate purification of the mRNA.

Comparison of in vitro ruminal fermentation incubated with different levels of Korean corn grains with total mixed ration as a basal

  • Hamid, Muhammad Mahboob Ali;Park, Ha Young;Choi, Chang Weon
    • Korean Journal of Agricultural Science
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    • v.45 no.3
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    • pp.419-427
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    • 2018
  • The present study was conducted to investigate the effect of different levels of Korean corn grain on in vitro ruminal fermentation with total mixed ration (TMR) as a basal feed. Three ruminal cannulated Holstein steers (Body Weight $479{\pm}33.0kg$) were used as rumen fluid donors. Treatments for in vitro fermentation were TMR only (control, 3.0 g), TMR substituted partially with high level (HC, TMR 1.5 and corn 1.5 g), and with low level of Korean corn grain (LC, TMR 2.25 and corn 0.75 g), respectively. To measure in vitro ruminal pH, gas production, ammonia N and volatile fatty acids (VFA), the in vitro fermentation incubation was triplicated at $39^{\circ}C$, 120 rpm for 0, 1, 3, 6, 12, 24 and 48 h, respectively. Mean ruminal pH was significantly lower (p < 0.05) for HC than control. Changes in rumen pH was rather similar between the groups till 6 h after incubation, but the lowest pH for HC (pH 5.10) appeared at 48 h compared with control and LC. Total gas production was tended (p < 0.09) to be higher and ammonia N was significantly lower (p < 0.05) for HC than control and LC. Total VFA was higher (p < 0.05) for HC and LC than control but no differences appeared between HC and LC. Overall, the present data indicate that feeding different levels of Korean domestic corn grain may lead to high and sustainable starch degradation in the rumen.

Protein Fractionation of Whole Crop Silages, and Effect of Borate-phosphate Buffer Extraction on In vitro Fermentation Characteristics, Gas Production and Degradation (사료작물 사일리지의 단백질 분획 및 Borate-phosphate Buffer 추출이 In vitro 발효성상, Gas 발생 그리고 분해율에 미치는 효과)

  • Shinekhuu, Judder;Jin, Guang-Lin;Ji, Byung-Ju;Li, Xiangzi;Oh, Young-Kyoon;Hong, Seong-Ku;Song, Man-Kang
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.369-378
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    • 2009
  • Protein fractionation was evaluated from whole crop silages of rye (RS), wheat (WS), triticale (TS), oat (OS), barley (BS), and rice straw silage (RSS), and in vitro trial was carried out to examine the effect of silage and extraction of soluble protein on fermentation characteristics, total gas production and degradation. Soluble protein of silages was extracted with borate-phosphate buffer, and fermentation characteristics, gas production and degradation of silages were estimated by incubating anaerobically the mixed solution of strained rumen fluid and artificial saliva (1:1, v/v) containing dried and ground silages placed in nylon bag at $39^{\circ}C$ up to 48h. Soluble protein (SP) content was lowest for RSS as 2.11% in total CP compared to those for other silages. Highest A fraction (NPN) was observed from RS (74.33% of total CP) while those from TS and RSS were relatively low (48%). B2 fraction was relatively higher for RS, RSS and WS than for TS and BS. $B_3$ fraction was lowest in WS among silages. C fraction (27.07) in RSS was higher than in other silages (1.40~9.93%). pH in incubation solution was increased (P<0.01~P<0.001) for extracted silages up to 12h but decreased (P<0.01) at 48h for non-extracted ones. Contents of ammonia-N (P<0.001) and total VFA (P<0.01~P<0.001) were higher for non-extracted silages than for extracted ones. Acetate proportion was increased (P<0.001) in buffer extracted silages while those of propionate and butyrate were decreased (P<0.001) up to 24h incubation. Increased (P<0.001) total gas production was obtained from non-extracted silages up to 12h while gas production was increased (P<0.01) in extracted ones thereafter. In vitro degradation of dry matter and CP was increased (P<0.001) in non-extracted silages but that of neutral detergent fiber was increased (P<0.001) in extracted ones without difference among silages. Difference in mean values of degradability for each silage prior to- and post extraction with borate buffer, however, was not found among silages. It may be concluded that high NPN content of silages may reduce the protein availability in silages and borate buffer soluble components in silages can stimulate the early stage of fermentation.

Effect of Fungal Elimination on Bacteria and Protozoa Populations and Degradation of Straw Dry Matter in the Rumen of Sheep and Goats

  • Li, D.B.;Hou, X.Z.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.1
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    • pp.70-74
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    • 2007
  • An in vitro study was carried out to investigate the differences in rumen microbes and fiber degradation capacity between sheep and goats. Three local male sheep and three Inner Mongolia male cashmere goats (aged 1.5 to 2 years; weight 25.0 to 32.0 kg) were each fitted with a permanent rumen cannula used to provide rumen fluid. Cycloheximide was used to eliminate rumen anaerobic fungi. The results showed that the quantities of fungal zoospores in the culture fluid of the control group were significantly greater in the sheep than in the goats; however, bacteria and protozoa counts were significantly higher in goats than in sheep. The digestibility of straw dry matter did not differ significantly between the two species before elimination of fungi, but tended to be higher for sheep (55.4%) than for goats (53.3%). The results also indicated that bacteria counts increased significantly after elimination of anaerobic fungi; however, the digestibility of straw dry matter significantly decreased by 12.1% and 8.6% for sheep and goats respectively. This indicated that the anaerobic fungi of the rumen played an important role in degradation of fiber.

Colon Delivery of Prednisolone Based on Chitosan Coated Polysaccharide Tablets

  • Park, Hyun-Sun;Lee, Jue-Yeon;Cho, Sun-Hye;Baek, Hyon-Jin;Lee, Seung-Jin
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.964-968
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    • 2002
  • Colon drug delivery is advantageous in the treatment of colonic disease and oral delivery of drugs unstable or suceptible to enzymatic degradation in upper GI tract. In this study, multilayer coated system that is resistant to gastric and small intestinal conditions but can be easily degraded by colonic bacterial enzymes was designed to achieve effective colon delivery of prednisolone. Variously coated tablets containing prednisolone were fabricated using chitosan and cellulose acetate phthalate (CAP) as coating materials. Release aspects of prednisolone in simulated gastrointestinal fluid and rat colonic extracts (CERM) were investigated. Also, colonic bacterial degradation study of chitosan was performed in CERM. From these results, a three layer (CAP/Chitosan/CAP) coated system exhibited gastric and small intestinal resistance to the release of prednisolone in vitro most effectively. The rapid increase of prednisolone in CERM was revealed as due to the degradation of the chitosan membrane by bacterial enzymes. The designed system could be used potentially used as a carrier for colon delivery of prednisolone by regulating drug release in stomach and the small intestine.

Effect of Ensiling with Acremonium Cellulase, Lactic Acid Bacterial and Formic Acid on Tissue Structure of Timothy and Alfalfa

  • Asian, Aniwaru;Okamoto, M.;Yoshihira, T.;Ataku, K.;Narasaki, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.10 no.6
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    • pp.593-598
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    • 1997
  • The changes of tissue structure in timothy and alfalfa during ensiling process with silage additives; lactic acid bacteria, cellulase and formic acid, were observed with a video microscope. Stem samples were obtained from the second internode, and cut to divide into 2 pieces. One piece was for observation of ensiled material and the other was for silage. The latter piece was put into a nylon cloth bag, and ensiled with grass for 50 days in a small experimental silo Lignification of the plant tissues was checked by acid phloroglucinol. Natural silage fermentation resulted in some degradation of less lignified parenchyma in both plant species. However, lignified sclerenchyma and vascular bundles remained intact. The cellulase enhanced the degradation of parenchyma tissue, while the formic acid suppressed the degradation. The effect of lactobacillus was small. The percentage of remained cross sectional area of stem and the loss of NDF and ADF by silage fermentation confirmed the observation. High negative correlations were obtained between the remained area and loss of fibrous components during silage fermentation in both plants, and between the loss of fibrous components and in vitro dry matter digestibility in timothy but not in alfalfa.

Effect of Vehicles and Enhancers on the in vitro Skin Penetration of Aspalatone and Its Enzymatic Degradation Across Rat Skins

  • Gwak, Hye-Sun;Chun, In-Koo
    • Archives of Pharmacal Research
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    • v.24 no.6
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    • pp.572-577
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    • 2001
  • The feasibility of skin penetration was studied for aspalatone (AM, acetylsalicylic acid maltol ester), a novel antithrombotic agent. In this studys hairless mouse dorsal skins were used as a model to select composition of vehicle and AM. Based on measurements of solubility and partition coefficient, the concentration of PC that showed the highest flux for AM across the hairless mouse skin was found to be 40%. The cumulative amount permeated at 48 h, however, appear inadequate, even when the PC concentration was employed. To identify a suitable absorption enhancer and its optimal concentration for AM, a number of absorption enhancers and a variety of concentration were screened for the increase in transdermal flux of AM. Amongst these, linoleic acid (LOA) at the concentration of 5% was found to have the largest enhancement factor (i.e., 132). However, a further increase in AM flux was not found in the fatty acid concentration greater than 5%, indicating the enhancement effect is in a bell-shaped currie. In a study of the effect of AM concentration on the permeation, there was no difference in the permeation rate between 0.5 and 1% for AM, below its saturated concentration. At the donor concentration of 2%, over the saturated condition, the flux of AM was markedly increased. A considerable degradation of AM was found during permeation studies, and the extent was correlated with protein concentrations in the epidermal and serosal extracts, and skin homogenates. In rat dorsal skins, the protein concentration decreased in the rank order of skin homogenate > serosal extract > epidermal extract. Estimated first order degradation rate constants were $6.15{\pm}0.14,{\;}0.57{\pm}0.02{\;}and{\;}0.011{\pm}{\;}0.004{\;}h^{-1}$ for skin homogenate, serosal extract and epidermal extract, respectively. Therefore, it appeared that AM was hydrolyzed to some extent into salicylmaltol by esterases in the dermal and subcutaneous tissues of skin. taken together, our data indicated that transdermal delivery of AM is feasible when the combination of PC and LOA is used as a vehicle. However, since AM is not metabolically stable, acceptable degradation inhibitors may be nervessary to fully realize the transdermal delivery of the drug.

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