• Title/Summary/Keyword: In vitro Incubation

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In vitro Nutrient Digestibility, Gas Production and Tannin Metabolites of Acacia nilotica Pods in Goats

  • Barman, K.;Rai, S.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.1
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    • pp.59-65
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    • 2008
  • Six total mixed rations (TMR) containing 0, 4, 6, 8, 10, 12% tannin (TMR I-VI), using Accacia nilotica pods as a source of tannin, were used to study the effect of Acacia tannin on in vitro nutrient digestibility and gas production in goats. This study also investigated the degraded products of Acacia nilotica tannin in goat rumen liquor. Degraded products of tannins were identified using high performance liquid chromatography (HPLC) at different hours of incubation. In vitro digestibility of dry matter (IVDMD) and organic matter (IVOMD) were similar in TMR II, and I, but declined (p<0.05) thereafter to a stable pattern until the concentration of tannin was raised to 10%. In vitro crude protein digestibility (IVCPD) decreased (p<0.05) with increased levels of tannins in the total mixed rations. Crude protein digestibility was much more affected than digestibility of dry matter and organic matter. In vitro gas production (IVGP) was also reduced (p<0.05) with increased levels of tannins in the TMR during the first 24 h of incubation and tended to increase (p>0.05) during 24-48 h of incubation. Gallic acid, phloroglucinol, resorcinol and catechin were identified at different hours of incubation. Phloroglucinol and catechin were the major end products of tannin degradation while gallate and resorcinol were produced in traces. It is inferred that in vitro nutrient digestibility was reduced by metabolites of Acacia nilotica tannins and ruminal microbes of goat were capable of withstanding up to 4% tannin of Acacia nilotica pods in the TMR without affecting in vitro nutrient digestibility.

Liquid Boar Sperm Quality during Storage and In vitro Fertilization and Culture of Pig Oocytes

  • Park, C.S.;Kim, M.Y.;Yi, Y.J.;Chang, Y.J.;Lee, S.H.;Lee, J.J.;Kim, M.C.;Jin, D.I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.10
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    • pp.1369-1373
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    • 2004
  • The percentages of sperm motility and normal acrosome on the liquid boar semen diluted and preserved at $4^{\circ}C$ with lactose hydrate, egg yolk and N-acetyl-D-glucosamine (LEN) diluent were significant differences according to preservation day and incubation time, respectively. The sperm motility steadily declined from 96.9% at 0.5 h incubation to 78.8% at 6 h incubation at 1 day of preservation. However, the sperm motility rapidly declined after 4 day of preservation during incubation. The normal acrosome steadily declined from 93.3% at 0.5 h incubation to 73.8% at 6 h incubation at 1 day of preservation. However, the normal acrosome rapidly declined after 3 day of preservation during incubation. The rates of sperm penetration and polyspermy were higher in 5 and $10{\times}10^6$ sperm/ml than in 0.2 and $1{\times}10^6$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in $10{\times}10^6$ sperm/ml compared with other sperm concentrations. The rates of blastocysts from the cleaved oocytes (2-4 cell stage) were highest in $1{\times}10^6$sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at $4^{\circ}C$ could be used for in vitro fertilization of pig oocytes matured in vitro. Also, we recommend $1{\times}10^6$sperm/ml concentration for in vitro fertilization of pig oocytes.

Comparison of In situ Dry Matter Degradation with In vitro Gas Production of Oak Leaves Supplemented with or without Polyethylene Glycol (PEG)

  • Ozkan, C. Ozgur;Sahin, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.8
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    • pp.1120-1126
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    • 2006
  • Dry matter (DM) degradation of leaves from Quercus cercis, Quercus libari, Quercus branti, and Quercus coccifera was determined using two different techniques: (i) in vitro gas production and (ii) the nylon bag degradability technique. In vitro gas production in the presence or absence of PEG and in situ DM disappearance were measured at 3, 6, 12, 24, 48, 72 and 96 h. In situ and in vitro DM degradation kinetics were described using the equation y = a+b ($1-e^{-ct}$). At all incubation times leaves from Quercus branti incubated with or without PEG gave significantly higher gas production than the other oak leaves except for 3 and 6 h incubation when leaves from Quercus branti without PEG supplementation only gave higher gas production than Quercus cercis and Quercus coccifera. At all incubation times except at 3, 6 and 12 h the DM disappearance from Quercus branti was significantly higher than the other species. Generally, PEG supplementation considerably increased the gas production at all incubation times and estimated parameters such as gas production rate ($c_{gas}$), gas production (ml) from the quickly soluble fraction ($a_{gas}$), gas production (b) from the insoluble fraction, potential gas production (a+b). However, all oak leaves did not give the same response to the PEG supplementation. Although the increase in gas production at 96 h incubation time was 8.9 ml for Quercus libari the increase was 5.5 ml for Quercus coccifera. It was concluded that except at early incubation times the relationships between the two methodologies seem to be sufficiently strong to predict degradability parameters from gas production parameters obtained in the presence or absence of PEG.

Effects of Corn Processing on In Vitro and In Situ Digestion of Corn Grain in Holstein Steers

  • Lee, S.Y.;Kim, W.Y.;Ko, J.Y.;Ha, J.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.6
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    • pp.851-858
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    • 2002
  • This study was conducted to determine effects of whole (intact), coarsely-ground (4 mm), finely-ground (1 mm), steam-flaked and steam-flaked-ground (1 mm) corns on in vitro and in situ DM digestibilities and also in vitro fermentation characteristics. After 48 h incubation, in vitro dry matter digestibilities of whole, steam-flaked, coarsely-ground, steam-flaked-ground, and finely-ground corns were 6.79, 61.68, 76.48, 85.72 and 90.31%, respectively. Steam-flaked-ground corn showed the highest digestibility until 24 h incubation (p<0.01). After 48 h incubation, pH of whole corn decreased with a small range. However the values of pH of other media significantly decreased (p<0.01). The gas productions of finely-ground and steam-flaked-ground corns were higher than those of the other corns (p<0.01). After 24 h incubation, $NH_3$-N concentration of finely-ground and steam-flaked-ground corns increased rapidly. Total VFA was the highest in finely-ground corn, followed by steam-flaked-ground, steam-flaked, coarsely-ground and whole corns. Incorporating steam-flaked corn resulted in the highest propionate concentration (p<0.01) and the lowest acetate : propionate value (p<0.05). Finely-ground corn showed the highest in situ DM digestibility throughout the incubation period (p<0.01), followed by coarsely-ground, steam-flaked and whole corns, respectively. Overall, DM of whole corn was merely digested in vitro as well as in situ.

An Estimation on the Efficiency of Utilization of Sulfur Coated Urea and Diuredio Isobutane in Vitro (In Vitro에 의한 유황피복요소(硫黃被覆尿素)와 Diuredio Isobutane의 이용효과(利用效果) 판정(判定))

  • Lee, Ho-Il;Rhee, Jae-Ku;Choi, In-Hyuk
    • Korean Journal of Veterinary Research
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    • v.22 no.2
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    • pp.111-114
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    • 1982
  • In order to explore more efficient protein substitutes by improving the utilization of non-protein nitrogen compounds in ruminants, the experiment was undertaken. The effects of sulfur coated urea (SCU) and diuredio isobutane (DUIB) in the ruminal fluid on the concentration of $NH_3-N$, the total count of the ruminal ciliates and pH value were estimated in vitro. The results obtained from the experiment were as follows: 1. The pH of the media of the group added diuredio isobutane and sulfur costed urea tends to decrease slightly at 9 hours after the incubation, but no pH changes were observed in the media added urea alone. 2. The number of the ciliates in the ruminal fluid was slightly increased 9 hours after the incubation in all groups. 3. The concentrations of $NH_3-N$ in the ruminal fluid were gradually increased according to the incubation in vitro showing 418, 431 and $627{\mu}g/ml$ in the group added diuredio isobutane and 428, 569 and $792{\mu}g/ml$ in the group added sulfur coated urea at 0, 0.5 and 9 hours after the incubation, respectively.

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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.

Comparison of in vitro ruminal fermentation between different originated corn grains

  • Kim, Do Hyung;Park, Ha Young;Choi, Chang Weon
    • Korean Journal of Agricultural Science
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    • v.44 no.4
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    • pp.541-548
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    • 2017
  • This study was conducted to compare in vitro rumen fermentation characteristics among corn grains imported from America, Brazil, Argentina and Ukraine A and Ukraine B. Two Holstein steers, each surgically fitted with a ruminal cannula, consuming total mixed ration were used as rumen fluid donors. In vitro rumen fermentation experiments were performed in a completely random design which included a control (no corn) and treatments with 3.0 g of corn from different geographical origins, i.e., America, Brazil, Argentina, and Ukraine A and Ukraine B, respectively. Ruminal pH, ammonia-N, volatile fatty acid (VFA) and total gas production were measured at 0, 1, 3, 6, 12, 24 and 48 h post-incubation, respectively. No differences (p > 0.05) in mean ruminal pH appeared among the treated groups, however, ruminal pH patterns differed; i.e. corn treated groups had dramatically lower pH compared with control during the entire incubation period. Similarly, no different patterns between the groups in ammonia-N (p > 0.05) appeared until 6 h post-incubation. Unexpectedly, higher ammonia-N concentration for control than that for the corn treated groups appeared after 12 h post-incubation despite that for all groups increased. Total VFA was similar between the groups until 6 h post-incubation, but VFA after 12 h post-incubation was different (p < 0.05), i.e. VFA for corn from Argentina, Ukraine A, Ukraine B, and Brazil were comparatively higher than for America. Overall, data in this study showed that the corns of different origins may have different feed values to ruminants despite having similar chemical compositions.

Effects of Supplementation of Spent Mushroom(Flammulina velutipes) Substrates on the in vitro Ruminal Fermentation Characteristics and Dry Matter Digestibility of Rye Silage (호밀 사일리지 제조 시 팽이버섯 수확 후 배지 첨가수준이 in vitro 반추위 발효특성 및 건물소화율에 미치는 영향)

  • Kang, Han-Byeol;Cho, Woong-Ki;Cho, Soo-Jeong;Lee, Shin-Ja;Lee, Sung-Sill;Moon, Yea-Hwang
    • Journal of agriculture & life science
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    • v.51 no.5
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    • pp.91-101
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    • 2017
  • The in vitro experiment was conducted to ensure the supplemental level of spent Flammulina velutipes mushroom substrates(SMS) as an energy source in manufacturing of rye silage. Rye harvested at heading stage was ensiled with spent mushroom substrates of 0%(Control), 20%(R-20), 40%(R-40) and 60%(R-60) as fresh matter basis for 6week. The rumen fluid for preparation of in vitro solution was collected from two cannulated Holstein bulls fed a 40:60 concentrate:timothy diet. The experiment was conducted by 3, 6, 9, 12, 24, and 48 hrs of ncubation time with 3 replications. The silages were evaluated fermentation characteristics and dry matter digestibility(DMD) in vitro. The pH of in vitro solution was inclined to decrease with elapsing the incubation time, and that of the R-60 was significantly(p<0.05) lower than the other treatment at 48 hr of incubation. The microbial growth in vitro was inclined to increase with elapsing the incubation time, and that of the R-20 was significantly(p<0.05) greater than the Control at 48 hr of incubation. Gas production was greater(p<0.05) in the Control than the other treatments at 48 hr of incubation. In vitro dry matter digestibility(IVDMD) was higher with increasing the supplemental level of SMS, and was significantly(p<0.05) lower in the Control compared with other treatments throughout whole incubation time. The IVDMD for R-60 was the highest(p<0.05) among treatments at 24 hr and 48 hr of incubation. Considering of above results and the availability of SMS, SMS could be supplemented by 60% in fresh matter basis for rye silage fermentation.

Effects of spent mushroom (Flammulina velutipes) substrates on in vitro ruminal fermentation characteristics and digestibility of whole crop sorghum silage (팽이버섯 수확후배지 첨가가 수수 사일리지의 in vitro 반추위 발효특성 및 소화율에 미치는 영향)

  • Moon, Yea Hwang;Chang, Sun Sik;Kim, Eun Tae;Cho, Woong Gi;Lee, Shin Ja;Lee, Sung Sil;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.3
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    • pp.163-169
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    • 2015
  • The in vitro experiment was conducted to ensure the supplemental level of spent Flammulina velutipes mushroom substrates (SMS) as an energy source in manufacturing of whole crop sorghum silage. Sorghum harvested at heading stage was ensiled with spent mushroom substrates of 20% (S-20), 40% (S-40) and 60% (S-60) as fresh matter basis for 6 week. The experiment was conducted by 3, 6, 9, 12, 24, 48 hrs of incubation time with 3 replications. The silages were evaluated fermentation characteristics and dry matter digestibility (DMD) in vitro. The pH of in vitro solution was inclined to decrease with elapsing the incubation time, and that of the S-20 was significantly (P<0.05) lower than the other treatment at 48 hr of incubation. Gas production was greater (P<0.05) in the S-20 than the other treatments at 6 and 12 hrs of incubation. The microbial growth in vitro was inclined to decrease following 24 hr of incubation, and thereafter sustained the similar levels. In vitro dry matter digestibility (IVDMD) was lowered by increasing the supplemental level of spent mushroom substrate, and was a low level in the S-60 throughout whole incubation time. Although the IVDMD for S-40 was steadily increased from 9 hr of incubation and reached to similar level with the S-20 at 48 hour of incubation, however SMS for whole crop sorghum silage fermentation might as well add about 20 to 30% in fresh matter basis when considering DMD.

Effects of Citrus Flavonoid, Hesperidin and Naringin on Lipid Metabolism in HepG2 Cells (간배양 HepG2 세포의 지질대사에 미치는 Hesperidin 및 Naringin의 영향)

  • 김범규;차재영;조영수
    • Journal of Life Science
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    • v.9 no.4
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    • pp.382-388
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    • 1999
  • The effects of citrus flavonoids, hesperidin and naringin, on the lipid metabolism were investigated in cultured human hepatocyte HePG2 cells. HepG2 cells were cultured for 6 h and 24 h to the control medium or the media containing hespridin and narigin, which concentrations were 0.5 and 5.0 mg/$m\ell$. There were no significant effects on cell proliferation and cellular protein content, except for increased in these parameters by adding both citrus flavonoids (0.5 mg/$m\ell$). The cellular content of triacylglycerol after 6 h incubation with 0.5 mg/$m\ell$ hesperidin and naringin was markedly increased, and after 24 h incubation that was decreased in both citrus flavonoids supplementation. The supplementation of 5.0 mg/$m\ell$ hesperidin caused a marked decrease in the cellular cholesterol content following 6 h incubation, and that was also reduced markdly, in a dose-dependent manner, during incubation for 24 h. However, there was no significant difference in the cellular cholesterol content in medium supplemented with naringin. The effect of hesperidin and naringin on acyl-CoA: cholesterol acyltransferase (ACAT) activity was studied in vivo and in vitro. The data confirmed that hesperidin inhibit ACAT activity in vivo and in vitro, whereas naringin had no such effect on ACAT activity in vivo but not in vitro. The present study suggests that hesperidin reduces the cellular triacyglycerol and cholesterol contents in human hepatocyte HepG2 cells.

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