• Parasites, Hosts and Diseases
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• 제35권1호
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• pp.25-30
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• 1997

참굴큰입흡충(GMmophQHoidesseoi)은 새로운 인체 기생 장흡충으로서 최근 우리 나라에서 인체 감염의 유행이 보고된 바 있다. 이 연구는 참굴큰입흡충의 피낭유충을 시험관내에서 배양하여 성충으로 발육시킬 수 있는지 알아보고자 시행하였다. 피낭유충은 자연산 감염 굴에서 회수하였고. NCTC 109 배지에 우태아 혈청(FBS) 20%와 항생제-항진균제(Gibco) 1%를 첨가하여 사용하되 $37^{\circ}C$/5%{\;}CO_2,{\;}41^{\circ}C/8%{\;}CO_2{\;}및{\;}41^{\circ}C/5%{\;}CO_2$의 3가지 배양조건 하에서 배양하였다 배양 2일에서 5일까지 각 조건 하에서 얻은 충체를 실험 감염 C3H 마우스에서 회수한 충체와 발육 및 성숙도를 비교하였다. 배양군의 충체 크기는 길이 $200-300{\;}\mu\textrm{m}$로서 피낭유충과 비교할 때 유의한 차이를 보이지 않았으나. 마우스로부터 회수한 충체는 $300-400{\;}\mu\textrm{m}$에서 유의한 차이를 보였다. 마우스 체내, $41^{\circ}C/5%{\;}CO_2{\;}및{\;}41^{\circ}C/8%{\;}CO_2$ 조건 하에서는 모두 2일째에 충란이 형성되기 시작하였으나, $37^{\circ}C/5%{\;}CO_2$ 하에서는 배앙 5일까지도 충란이 형성되지 않았다 배양군 중에서는 $41^{\circ}C/5%{\;}CO_2$ 조건이 충체 성숙 및 충란 생산에 가장 적합하였으나 마우스 체내에서 자란 충체에 비해 자궁내 충란 수가 약 1/2에 불과하였다. 결론적으로 참굴큰입흡충의 시험관내 배양은 $41^{\circ}C/5%{\;}CO_2$ 또는 $41^{\circ}C/8%{\;}CO_2$ 조건에서 부분적으로 성공적임을 알 수 있었다.

• Parasites, Hosts and Diseases
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• 제48권1호
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• pp.49-55
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• 2010

In vitro cultivation of trematodes would assist studies on the basic biology of the parasites and their hosts. This is the first study to use the yolk of unfertilized chicken eggs as a simple and successful method of ovocultivation and the first time to obtain the adult-stage of the trematode Cymatocarpus solearis Braun, 1899 (Digenea: Brachycoeliidae). Chicken eggs were inoculated with metacercariae from the muscle of the spiny lobster, Panulirus argus (Latreille, 1804). The metacercariae were excysted and incubated for 576 hr (24 days) at $38^{\circ}C$ to obtain the adult stage. Eggs in utero were normal in shape and light brown color. The metacercariae developed into mature parasites that have been identified as the adultstage found in marine turtles. The adult lobsters collected in Quintana Roo State, Mexico, showed the prevalence of 49.4% and the mean intensity of 26.0 per host (n=87). A statistical study was performed to determine that no parasitic preference was detected for male versus female parasitized lobsters. Morphometric measurements of the adult-stage of C. solearis obtained in our study have been deposited in the National Helminths Collection of the Institute of Biology of the National Autonomous University of Mexico. This study is significant because it is the first time that a digenean of the family Brachycoeliidae has been demonstrated to develop in vitro from metacercariae into adults capable of producing eggs using the yolk of unfertilized chicken eggs. Secondly, this technique allows to obtain the adult stage of C. solearis without the presence of its marine turtle host, allows us to describe the mature parasites, and thus contribute to our understanding of the biology of C. solearis.

• Plant Resources
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• 제8권1호
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• pp.48-51
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• 2005

Optimmum cultivation period was determined for producing seed bulb of Korean native Allium wakegi Araki in vitro in hydroponic culture. The growth gradually increased during cultivation period. In general, plants grown for 5 months produced significantly the highest bulb number and bulb fresh weight per plant. Raising the cultivation period from 1 to 5 months remarkably increased seed bulb yield.

• Asian-Australasian Journal of Animal Sciences
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• 제24권11호
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• pp.1541-1546
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• 2011

This study was carried out to examine the influence of minimum essential medium (MEM) vitamins supplementation to in vitro maturation medium and in vitro culture medium on the development of porcine embryos. Porcine embryo development was investigated following cultivation in both in vitro maturation and culture medium with the supplementation of MEM vitamins (0, 0.1, 0.2 and 0.4%) using immature oocytes collected from the ovary of prepubertal gilts. Embryo development was observed and the total cell number in each blastocyst generated under the culture conditions was quantified following supplementation of the medium. The embryonic development rate of the blastocyst and hatched blastocyst was higher, but not significantly so, when 0.4% MEM vitamins were supplemented to the in vitro maturation medium of the porcine oocyte. Interestingly, the total number of cells in the blastocyst was significantly higher in the in vitro maturation MEM vitamins supplemented group compared to either the untreated group or the group which had MEM vitamins supplemented to both in vitro maturation and in vitro culture medium simultaneously (p<0.05). Therefore, the supplementation of 0.4% MEM vitamins to the in vitro mature medium has a beneficial effect on the embryonic development of in vitro produced blastocysts derived from the immature porcine oocytes.

• Journal of the Korean Wood Science and Technology
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• 제33권5호통권133호
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• pp.92-98
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• 2005

This study was performed to get the basic information concerned to the optimum culture condition of Inonotus obliquus. Several solid media, PDA, MEA and Czapek-Dox, and three liquid media were adopted for the in vitro cultivation. Some main features of the fungal morphological characteristics under cultivation conditions were observed and described. Preliminary results showed that appearance of the mycelial mat, hyphal size and substrate pigmentation differed according to the media. The PDA medium was the most favorable substrate for the growth on solid culture, followed by MEA and Czapek-Dox media. Concerned to the addition of amino acids, 5 amino acids, such as alanine, alginine, isoleucine, leucine and threonine, enhanced to the mycelial growth. Isoleucine was shown the best fungal growth. An important morphological hyphal structure for the fungus, the setae, was found in abundance and diverse its shape and size. In liquid culture, fresh potato broth was the best growth stimulant of the fungus, followed by Malt extract and potato broth. Addition of yeast extract to the liquid media had improved the biomass, but not laccase production.

• Journal of Plant Biotechnology
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• 제42권2호
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• pp.111-116
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• 2015

Interest and great demand for blueberry (Vaccinium corymbosum) have increased, as V. corymbosum is now one of the most economically important crops in Korea. It is expected that blueberry production and the area planted for cultivation will increase consistently in the years ahead because of high profitability and the consumer's demand for healthy ingredients. Effective mass production of blueberry is urgently needed for commercial cultivation establishment, but a main limitation is lack of a propagation system that produces a disease-free plant material for commercial plantation. A large amount of research has focused entirely on developing tissue culture techniques for blueberry propagation. However, controlling fungal and bacterial contamination of woody plant material is extremely difficult. Our study was conducted to investigate the effect of biocide addition during the in vitro culture of blueberry on plantlet growth and contamination occurrence. Four biocides, including Plant Preservative Mixture ($PPM^{TM}$), vancomycin, nystatin and penicillin G, were used in varying concentrations during the in vitro propagation of blueberry. When nystatin was added into the medium at low concentrations, the overall growth of blueberry plantlets was retarded. Addition of vancomycin and penicillin G in high concentrations decreased contamination but induced plantlet mortality. On the other hand, when 1ml/L $PPM^{TM}$ was added, the growth characteristics of blueberry plantlets did not significantly differ from non-treatment (control), and the contamination occurrence rate was very low. From these results, we found that the addition of the appropriate biocide could provide an effective method to reduce contamination in the culture process, thereby raising in vitro production efficiency.

• 대한미생물학회지
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• 제7권1호
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• pp.29-41
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• 1972

조직배양(組織培養)을 이용(利用)한 마우스복강거식세포내 인나균증식실험(人癩菌增殖實驗)으로서 trypsin-정제인나균(精製人癩菌)을 사용하여 1) 인나균(人癩菌)의 마우스복강내접종(腹腔內接種)에 의(依)하여 야기(惹起)되는 복강거식세포내의 생체내(生體內) 인나균감염(人癩菌感染)에 대(對)한 동역학적양상(動力學的樣相), 2) in vivo infection-in vitro cultivation에 의(依)한 인나균(人癩菌)의 복강거식세포내 증식태도(增殖態度) 그리고 3) 인나균(人癩菌)의 복강내접종(腹腔內接種)으로 인(因)한 마우스비장조직(脾臟組織)의 병리학적변화(病理學的變化)를 구명(究明)코저 본연구(本硏究)를 실시(實施)하였으며 아래의 결론(結論)을 얻었다. 1. 인나균(人癩菌)의 마우스복강내접종후(腹腔內接種後) 복강거식세포배양실시까지의 기간(其間)이 연장(延長)됨에 따라 배양(培養)된 복강거식세포에 있어서 세포질내(細胞質內)에 식균된 항산균수(抗酸菌數)와 항산균(抗酸菌)을 식균한 거식세포수(細胞數)가 각각 계속적(繼續的)으로 현저(顯著)하게 감소(減小)되어감이 관찰(觀察)되었다. 2. 인나균(人癩菌)의 복강내접종후(腹腔內接種後) 5개월(個月)에 이르기까지 생체내(生體內)에 존재(存在)하는 마우스복강거식세포에 있어서의 인나균증식(人癩菌增殖)을 관찰(觀察)할 수 없었다. 3. 인나균(人癩菌)의 복강내접종후(腹腔內接種後) 24시간(時間) 및 1주(週)만에 실시(實施)된 복강거식세포배양을 2 내지(乃至) 3개월간(個月間) 그 배양상태(培養狀態)를 유지(維持)하였던바 배양(培養)된 거식세포(細胞)의 염색표본(染色標本)에서 거식세포내(細胞內) 항산균수(抗酸菌數)의 뚜렷한 증가양상(增加樣狀)을 관찰(觀察)할수 있었다. 4. in vivo infection-in vitro cultivation 수기(手技)로서 배양(培養)된 복강거식세포를 사용(使用)한 총항산균수측정실험(總抗酸菌數測定實驗)에서 조직배양개시(組織培養開始) 9 및 11주(週)에 이르러 배양(培養)된 거식세포내(細胞內) 항산균수증가(抗酸菌數增加)에 대한 양적증거(量的證據)를 얻을수 있었다. 5. 인나균(人癩菌)의 복강내접종(腹腔內接種)으로 야기되는 마우스비장조직(脾臟組織)의 병리학적소견(病理學的所見)은 주로 적수부위(赤髓部位)에 나타난 변성변화(變性變化)이었으며, 균접종후(菌接種後) 5개월(個月)에 이르기까지 마우스비장내(脾臟內)에 있어서의 인나균(人癩菌)의 증식양상(增殖樣狀)을 관찰(觀察)할 수 없었다.

• 대한수의학회지
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• 제38권1호
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• pp.107-117
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• 1998

The cultivation for development of Ascaris suum from the second-stage larvae($L_2$) embryonated egg and the third-stage of rat-derived larvae($L_3$) recovered from lung of rats were performed to use the screening test of anthelmintics in vitro. The preparations of larvae for cultivation were that the artificially-hatched $L_2$ incubated the embryonated eggs of Ascaris suum in 0.1% formalin solution at $25^{\circ}C$ for 28 days and the rat-derived larvae($L_3$) recovered from the lung of rat infected with the embryonated eggs of Ascaris suum on 7 days after infection(DAI). The cultivation for development of Ascaris suum from the embryonated eggs($L_2$) and the rat-derived larvae($L_3$) for 14 days in RPMI medium 1640(with 5% bovine calf serum) were as follows : 1. The sizes of the liberated larvae($L_2$) which were artificially hatched from embryonated eggs with glass beads(diameter 5mm) were $190{\sim}250{\mu}m$ on 1 days in culture(DIC). The second-stage larvae were molted into third-stage larvae(early $L_3$; $250{\sim}300{\mu}m$) and the features of these larvae were first observed such as cephalic cuticle, esophageal lumen and anus etc. on 5 DIC and the sizes of late third-stage larvae were $250{\sim}450{\mu}m$ on 10 DIC. The sizes of early fourth-stage larvae($L_4$) were $500{\sim}700{\mu}m$ and the features of these larvae were more pronounced in internal organs on 15 DIC. 2. The sizes of third-stage larvae($L_3$) recovered from the lung of rats were $1,340{\sim}1,370{\mu}m$ and the feartures of cephalic cuticle, esophageal lumen, intestine, rectum, anus were visualized by inverted microscope on 1 DIC. The fourth-stage larva($L_4$) completed by third ecdysis were recognizable and sizes of early fourth-stage larvae were developed as $1,400{\sim}2,200{\mu}m$ on 5 DIC. The sizes of middle fourth-stage of larva were $1,900{\sim}2,300{\mu}m$ and the thickened epithelial rectum was observed on 10 DIC. The rectum and anus of late fourth-stage larva($L_4$ $2,500{\sim}3,200{\mu}m$) had developed completely in RPMI medium 1640 on 15 DIC.

• 한국가축번식학회지
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• 제18권1호
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• pp.47-54
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• 1994

The objective of this study was to produce Korean native calves following transfer of in vitro matured, fertilized and cultured embryos into Holstein cows. The ovaries of Korean native cows or heifers were obtained from an abattoir and kept on 25 to 28$^{\circ}C$ and transported to laboratory within 2 hrs. The oocytes were matured in vitro (IVM) for 24 hrs in TCM-199 supplemented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, 1$\mu\textrm{g}$/ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% CO2 in air. They were fertilized in vitro (IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro (IVC) with bovine oviductal epithelial cells for 7 to 9 days. Late morulae and blastocysts produced in vitro were nonsurgically transferred to recipient cows by unilaterial. Recipients were monitored for estrus and for pegnancy by rectal plapation in 60 days after embryo transfer. One of them was pregnant to term and produced a female weighing 42.5kg at birth.

• Asian-Australasian Journal of Animal Sciences
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• 제32권1호
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• pp.103-109
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• 2019

Objective: Ensiling of tannin-rich fruit byproducts (FB) involves quantitative and qualitative changes in the tannins, which would consequently change the rumen fermentation characteristics. This study aimed to evaluate whether ensiled FBs are effective in mitigating methane emission from ruminants by conducting in vitro assessments. Methods: Fruit byproducts (grape pomace, wild grape pomace, and persimmon skin) were collected and subjected to four-week ensiling by Lactobacillus buchneri inoculant. A defined feed component with or without FB samples (both fresh and ensiled material) were subjected to in vitro anaerobic culturing using rumen fluid sampled from beef cattle, and the fermentation parameters and microbial populations were monitored. Results: Reduced methane production and a proportional change in total volatile fatty acids (especially enhanced propionate proportion) was noted in bottles containing the FBs compared with that in the control (without FB). In addition, we found lower gene copy number of archaeal 16S rRNA and considerably higher levels of one of the major fibrolytic bacteria (Fibrobacter succinogenes) in the bottles containing FBs than in the control, particularly, when it was included in a forage-based feed. However, in the following cultivation experiment, we observed that FBs failed to exhibit a significant difference in methane production with or without polyethylene glycol, implying that tannins in the FBs may not be responsible for the mitigation of methane generation. Conclusion: The results of the in vitro cultivation experiments indicated that not only the composition but also ensiling of FBs affected rumen fermentation patterns and the degree of methane generation. This is primarily because of the compositional changes in the fibrous fraction during ensiling as well as the presence of readily fermented substrates, whereas tannins in these FBs seemed to have little effect on the ruminal fermentation kinetics.