• Title/Summary/Keyword: in vitro production

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The Effect of Yerba Mate (Ilex Paraguariensis) Supplementation on Nutrient Degradability in Dairy Cows: An In sacco and In vitro Study

  • Hartemink, Ellen;Giorgio, Daniela;Kaur, Ravneet;Di Trana, Adriana;Celi, Pietro
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1606-1613
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    • 2015
  • This study was carried out to investigate the effects of Yerba Mate (YM) supplementation on nutrients' degradation, in vitro dry matter disappearance, gas production and rumen ammonia concentration. Three rumen-fistulated Holstein Friesian cows were used for the in situ incubations and provided rumen liquor for in vitro incubations. The inclusion of YM in a control diet (pasture+pellets) affected some in sacco degradation parameters. YM supplementation decreased the effective degradability and degradation rate of pasture crude protein (CP), and it seems to slow down the degradation of pasture neutral detergent fiber. A significant increase of degradation of pasture acid detergent fiber (ADF) was detected after YM inclusion in the control diet. YM supplementation reduced in vitro gas production of pasture and ammonia concentration of pellets. The addition of YM in ruminant diet could decrease ammonia production and increase protein availability for productive purposes. The moderate presence of tannins in YM could have affected the degradation kinetics of pasture CP and ADF and the ammonia production of pellets.

In Vitro Growth and Development of Mouse Preantral Follicles

  • Kim, Dong-Hoon
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.9-9
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    • 2000
  • The mammalian ovary has a large number of primordial and preantral follicles, which are a potential source of oocytes for the in vitro mass production of embryos. Several in vitro culture systems have been developed to support the growth and development of oocytes from mouse preantral follicles. Under the appropriate condition, meiotically incompetent oocytes from preantral follicles can grow to final size and complete nuclear maturation in vitro. Furthermore, the successful production of live young from in vitro grown and matured oocytes demonstrates that oocytes from preantral follicles are able to acquire full developmental capacity in vitro. However, the efficiency of in vitro production of embryos from mouse preantral follicles is still low. In farm animals as well as human, the growth of oocyte from preantral follicle to the meiotic competence stage has yet to be demonstrate. Therefore, further studies to improve the culture condition or to develope new culture system should be needed in the future. In addition, the visible progress in the establishment of the in vitro culture system for preantral follicles of farm animals and human could help to enlarge the populations of valuable agricultural, phamaceutical product-producing, and endangered animals, and to rescue the oocytes of women about to undergo clinical procedures that jeopardize oocytes.

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Comparison of In vivo and In vitro Techniques for Methane Production from Ruminant Diets

  • Bhatta, Raghavendra;Tajima, K.;Takusari, N.;Higuchi, K.;Enishi, O.;Kurihara, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1049-1056
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    • 2007
  • This study was conducted to compare the methane ($CH_4$) production estimated by in vivo (sulfur hexafluoride tracer technique ($SF_6$)) with that of two in vitro rumen simulation (RUSITEC) and gas production (IVGPT)) techniques. Four adult dry Holstein cows, aged $7.4{\pm}3.0$ years and weighing $697{\pm}70$ kg, were used for measuring methane production from five diets by the $SF_6$ technique. The experimental diets were alfalfa hay ($D_1$), corn silage + soybean meal (SBM) (910: 90, $D_2$), Italian rye grass hay +SBM (920: 80, $D_3$), rice straw +SBM (910: 90, $D_4$) and Sudan grass hay +SBM (920: 80, $D_5$). Each diet was individually fed to all 4 cows and 5 feeding studies of 17 d each were conducted to measure the methane production. In the RUSITEC, methane production was measured from triplicate vessels for each diet .In vitro gas production was measured for each of the diets in triplicate syringes. The gas produced after 24 and 48 h was recorded and gas samples were collected in vacuum vials and the methane production was calculated after correction for standard temperature and pressure (STP). Compared to the $SF_6$ technique, estimates of methane production using the RUSITEC were lower for all diets. Methane production estimated from 24 h in vitro gas production was higher (p<0.001) on $D_1$ as compared to that measured by $SF_6$, whereas on $D_2$ to $D_5$ it was lower. Compared to $SF_6$, methane production estimated from 48 h in vitro gas production was higher on all diets. However, methane estimated from the mean of the two measurement intervals (24+48 h/2) in IVGPT was very close to that of $SF_6$ (correlation 0.98), except on $D_1$. The results of our study confirmed that IVGPT is reflective of in vivo conditions, so that it could be used to generate a database on methane production potential of various ruminant diets and to examine strategies to modify methane emissions by ruminants.

Factors Influencing the Efficiency of In Vitro Embryo Production in the Pig

  • Lin, Tao;Lee, Jae Eun;Shin, Hyun Young;Oqani, Reza K.;Jin, Dong Il
    • Reproductive and Developmental Biology
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    • v.39 no.2
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    • pp.29-36
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    • 2015
  • Pigs are considered an ideal source of human disease model due to their physiological similarities to humans. However, the low efficiency of in vitro embryo production (IVP) is still a major barrier in the production of pig offspring with gene manipulation. Despite ongoing advances in the associated technologies, the developmental capacity of IVP pig embryos is still lower than that of their in vivo counterparts, as well as IVP embryos of other species (e.g., cattle and mice). The efficiency of IVP can be influenced by many factors that affect various critical steps in the process. The previous relevant reviews have focused on the in vitro maturation system, in vitro culture conditions, in vitro fertilization medium, issues with polyspermy, the utilized technologies, etc. In this review, we concentrate on factors that have not been fully detailed in prior reviews, such as the oocyte morphology, oocyte recovery methods, denuding procedures, first polar body morphology and embryo quality.

Improvement of Nutritive Value and In vitro Ruminal Fermentation of Leucaena Silage by Molasses and Urea Supplementation

  • Phesatcha, K.;Wanapat, M.
    • 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.

Production of Mycoplasma-Free Jujube Trees through In vitro Micrografting (기내미세접목(器內微細接木)에 의(依)한 대추나무의 마이코플라즈마 무병주(無病株) 생산(生産))

  • Park, Jae In
    • Journal of Korean Society of Forest Science
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    • v.82 no.3
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    • pp.254-259
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    • 1993
  • For the production of mycoplasma-free jujube trees from mycoplasma-infected trees in vitro micrografting visas carried out using apical meristems of in vitro grown plantlets as scions. The rootstocks were hypocotyl segments of in vitro germinated seedlings of Zizyphus jujuba Mill. which is widely used as rootstocks in the field grafting. Ten percent of scions showed normal growth and grew into plants. The presence of mycoplasma was tested using transmission electron microscopy and fluorescence microscopy. Mycoplasma was not found in the tissues of scion parts and seedlings, whereas it was found in in vitro grown plantlets. This suggests that the production of mycoplasmaa-free jujube trees is possible by the in vitro micrografting technique.

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Indirect Estimation of CH4 from Livestock Feeds through TOCs Evaluation

  • Kim, M.J.;Lee, J.S.;Kumar, S.;Rahman, M.M.;Shin, J.S.;Ra, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.4
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    • pp.496-501
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    • 2012
  • Thirty-five available feeds were fermented in vitro in order to investigate their soluble total organic carbon (TOCs) and methane ($CH_4$) production rate. A fermentation reactor was designed to capture the $CH_4$ gas emitted and to collect liquor from the reactor during in vitro fermentation. The results showed that $CH_4$ production rate greatly varied among feeds with different ingredients. The lowest $CH_4$-producing feeds were corn gluten feed, brewer's grain, and orchard grass among the energy, protein, and forage feed groups, respectively. Significant differences (p<0.05) were found in digestibility, soluble total organic carbon (TOCs), and $CH_4$ emissions among feeds, during 48 h of in vitro fermentation. Digestibility and TOCs was not found to be related due to different fermentation pattern of each but TOCs production was directly proportional to $CH_4$ production (y = 0.0076x, $r^2$ = 0.83). From this in vitro study, TOCs production could be used as an indirect index for estimation of $CH_4$ emission from feed ingredients.

Effects of lactic acid bacteria and molasses on fermentation dynamics, structural and nonstructural carbohydrate composition and in vitro ruminal fermentation of rice straw silage

  • Zhao, Jie;Dong, Zhihao;Li, Junfeng;Chen, Lei;Bai, Yunfeng;Jia, Yushan;Shao, Tao
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.783-791
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    • 2019
  • Objective: This study was to evaluate the fermentation dynamics, structural and nonstructural carbohydrate composition and in vitro gas production of rice straw ensiled with lactic acid bacteria and molasses. Methods: Fresh rice straw was ensiled in 1-L laboratory silos with no additive control (C), Lactobacillus plantarum (L), molasses (M) and molasses+Lactobacillus plantarum (ML) for 6, 15, 30, and 60 days. After storage, the silages were subjected to microbial and chemical analyses as well as the further in vitro fermentation trial. Results: All additives increased lactic acid concentration, and reduced pH, dry matter (DM) loss and structural carbohydrate content relative to the control (p<0.05). The highest organic acid and residual sugar contents and lignocellulose reduction were observed in ML silage. L silage had the highest V-score with 88.10 followed by ML silage. L and ML silage improved in vitro DM digestibility as compared with other treatments, while in vitro neutral detergent fibre degradability (IVNDFD) was increased in M and ML silage (p<0.05). M silage significantly (p<0.05) increased propionic acid (PA) content and decreased butyric acid content and acetic acid/PA as well as 72-h cumulative gas production. Conclusion: The application of ML was effective for improving both the fermentation quality and in vitro digestibility of rice straw silage. Inclusion with molasses to rice straw could reduce in vitro ruminal gas production.

Relative Palatability to Sheep of Some Browse Species, their In sacco Degradability and In vitro Gas Production Characteristics

  • Abdulrazak, S.A.;Nyangaga, J.;Fujihara, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1580-1584
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    • 2001
  • A study was conducted to estimate the nutritive value of some selected acacia forages using palatability index, in sacco degradability and in vitro gas production characteristics. Ten wethers (mean wt. $18{\pm}3.5kg$) were offered Acacia tortilis, Acacia nilotica, Acacia mellifera, Acacia brevispica, Acacia Senegal and Leucaena leucocephala (control) using a cafeteria system to determine the species preference by the animals. The acacia species were rich in nitrogen and showed variable palatability pattern. Significant (p<0.05) differences in relative palatability index (RPI) were detected among the species with the following ranking: brevispica > leucaena > mellifera > tortilis > Senegal > nilotica. Acacia nilotica appeared to be of low relative palatability with RPI of 24% and this was attributed to relatively high phenolic concentrations. The DM potential degradability (B) and rate of degradation (c) of the species were significantly (p<0.05) different, ranging from 40.1 to 59.1% and 0.0285 to 0.0794/h respectively. Acacia species had moderate levels of rumen undegradable protein, much higher than that in leucaena. In vitro gas production results indicated the effect of polyphenolic compounds on the fermentation rate, with lower gas production recorded from A. nilotica and tortilis. Based on RPI, A. brevispica and mellifera were superior to the rest and comparable to L. leucocephala. Long-term feeding trials are required with the superior species when used as protein supplements to poor quality diets.

Effect of Crude Protein Levels in Concentrate and Concentrate Levels in Diet on In vitro Fermentation

  • Dung, Dinh Van;Shang, Weiwei;Yao, Wen
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.6
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    • pp.797-805
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    • 2014
  • The effect of concentrate mixtures with crude protein (CP) levels 10%, 13%, 16%, and 19% and diets with roughage to concentrate ratios 80:20, 60:40, 40:60, and 20:80 (w/w) were determined on dry matter (DM) and organic matter (OM) digestibility, and fermentation metabolites using an in vitro fermentation technique. In vitro fermented attributes were measured after 4, 24, and 48 h of incubation respectively. The digestibility of DM and OM, and total volatile fatty acid (VFA) increased whereas pH decreased with the increased amount of concentrate in the diet (p<0.001), however CP levels of concentrate did not have any influence on these attributes. Gas production reduced with increased CP levels, while it increased with increasing concentrate levels. Ammonia nitrogen ($NH_3$-N) concentration and microbial CP production increased significantly (p<0.05) by increasing CP levels and with increasing concentrate levels in diet as well, however, no significant difference was found between 16% and 19% CP levels. Therefore, 16% CP in concentrate and increasing proportion of concentrate up to 80% in diet all had improved digestibility of DM and organic matter, and higher microbial protein production, with improved fermentation characteristics.