• Title/Summary/Keyword: B-2 cell

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Effects of Deer Antler on the Regeneration of Peripheral Nerves; About Sprout Formation of Experimentally Transected Sciatic Nerves in Rat (말초신경의 재생에 대한 녹용의 효과; 랫드에서 실험적 절단 좌골신경의 Sprout 형성에 관해)

  • Chang, Byung-Joon;Cho, Ik-Hyun;Choi, Hye-Young;Won, Hui-Young;Park, Chang-Hyun;Bae, Chun-Sik;Choe, Nong-Hoon
    • Applied Microscopy
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    • v.32 no.1
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    • pp.67-80
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    • 2002
  • This study was carried out to investigate the effects of deer antler extract on the regeneration of peripheral nerves. Sprague-Dawley male rats weighing about 300 gm were fed deer antler extract for 1, 2, and 3 weeks per oral (1.5 ml/100 gm B.W.), respectively, once a day and transected both sides of sciatic nerve of each leg. After keeping for 6 hours, sciatic nerves taken from proximal part of transected region were treated with conventional transmission electron microscopical method and then observed with electron microscope. The results obtained were summarized as follows; 1. Sciatic nerves of normal control group were not showing any sprouts and electron dense axolemmal projections were frequently observed. 2. Sciatic nerves of saline treated groups were showing axonal sprouts at the nodes of Ranvier. The length of them was usually short, and numerous vesicles, vacuoles and organelles including neurofilament were contained. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 29 (29%) in 1 week treated group, 32 (32%) in 2 weeks treated group, and 30 (30%) in 3 weeks treated group, respectively. 3. Sciatic nerves of deer antler treated groups were showing axonal sprouts at the node of Ranvier as well. Although most of the sprouts were short, some sprouts of 2 weeks and 3 weeks treated groups were quite long. Sprouts usually contained numerous vesicles, vacuoles and cell organelles such as neurofilaments and mitochondria. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 38 (38%) in 1 week treated group, 46 (46%) in 2 weeks treated group, and 48 (48%) in 3 weeks treated group respectively. The results described above explain pretreatment of deer antler extract improves the sprout formation of transected sciatic nerves, and then it suggests deer antler may be effective for the regeneration of peripheral nerves.

Prediction of Parthenogenetic Developmental Potential by Polar Body Extrusion and First Cleavage on In Vitro Maturation and Development of Porcine Follicular Oocytes (돼지 난모 세포의 Ethanol 처리에 의한 단위 발생에 있어서 극체 방출란과 분할란 선별에 따른 배발달율 비교)

  • Kim, H.J.;Cho, S.R.;Choe, J.Y.;Choi, S.H.;Han, M.H.;Son, D.S.;Lee, S.S.;Sang, B.D.;Ryu, I.S.;Kim, I.C.;Kim, S.J.;Kim, I.H.;Kim, S.K.
    • Journal of Embryo Transfer
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    • v.22 no.2
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    • pp.121-126
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    • 2007
  • The objective of this study was carried out to examine the selection effects of in vitro matured porcine follicular oocytes with polar body extrusion and early cleavage as non-invasive marker to know the developmental competence in advance. The porcine oocytes matured for 48 h were examined the polar body extrusion. The examined oocytes were matured for additional $16{\sim}18h$ and activated with 7% ethanol and cultured in $5{\mu}g/ml$ cytochalasin B for 5 h for diploid formation. The treated oocytes were cultured and examined the cleavage after 48 h and continued culturing for 5 days. The oocytes of 21.9% were discarded in morphological selection and 32.1% oocytes were discarded by failure of first polar body extrusion. The selected oocytes were matured and activated and then after 48 h the cleavage rates were examined. In morphologically selected oocytes, 15.8% oocytes were not cleaved and 52.6% oocytes were normally cleaved and 31.6% oocytes were hyper-cleaved over 8-cell stage. However in the first polar body extruded oocytes, 7.1% oocytes were not cleaved and 73.1% oocytes were normally cleaved and 19.8% oocytes were hyper-cleaved. The morphologically selected embryos that not cleavage-selected were developed in 16.7% up to blastocyst and the morphologically selected and cleavage-selected embryos were developed in 31.7%. The polar body extruded oocytes that were not carried out cleavage selection were developed in 39.0% and the polar body extruded and cleavage-selected embryos were developed 49.0%. The first cleavage timing was examined with 12 h interval after activation. In $0{\sim}12,\;12{\sim}24,\;24{\sim}36,\;and\;36{\sim}48h$ intervals, 4.1%, 68.6%, 19.1%, and 2.3% oocytes were cleaved and 5.9% oocytes were not cleaved until 48 after activation. The cleaved oocytes in each interval were cultured and developed upto blastocyst with 0, 39.1, 9.5, and 0%, respectively. This results suggests that polar body extruded and cleaved at $12{\sim}36h$ embryo has higher developmental potential than the others.

Biological Characters of Bacillus pumilus CPB-St Inhibiting the Growth of Fish Pathogenic Streptococci (어류병원성 연쇄구균의 생장을 억제하는 Bacillus pumilus CPB-St의 생물학적 특성)

  • Lee, Minyeong;Kim, Eunheui
    • Journal of fish pathology
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    • v.28 no.2
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    • pp.63-69
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    • 2015
  • The biological characteristics of Bacillus sp.CPB-St as a probiotic strain to control fish streptococcosis was determined. Based on 16S rRNA sequencing, Bacillus sp.CPB-St was identified as Bacillus pumilus and named B. pumilus CPB-St (Abbreviated as CPB-St). Growth inhibitory activity of CPB-St against Streptococcus spp. was examined at three different incubation temperatures ($20^{\circ}C$, $25^{\circ}C$, and $30^{\circ}C$) and three culture media (NA, TSA, and BHIA) based on the diameter of inhibition zone. Its activity (inhibition zone of 11~29 mm) at $20^{\circ}C$ was higher than that (12~21 mm) at $30^{\circ}C$. Its activity (29 mm) in NA media was the same as that (29 mm) in TSA media. However, it was higher than that (22 mm) in BHIA media. The inhibitory activity of CPB-St against Streptococcus spp. was high at pH7. However, its activity was the same at salinity of 0.5% to 3%. CPB-St showed maximum growth after incubation at $25^{\circ}C$ for 48 h. To use CPB-St as probiotics, settlement studies in fish intestine and its efficacy through feeding are needed. CPB-St was highly resistant to gastric juice at pH4 and flounder's bile salt as well as deoxycholic acid at $300{\mu}g/ml$. CPB-St showed optimal viability in 1% NaCl. It showed similar growth in 0% to 7% NaCl. CPB-St could tolerate $-20^{\circ}C$ and $-70^{\circ}C$ for 45 min. There was no difference in the growth of the strain between room temperature and $4^{\circ}C$. Fish diet supplemented with CPB-St could be stored at low temperature without cell loss. Therefore, CPB-St might be used as probiotics to control streptococcosis of fish.

Enhancement of Anti-inflammatory Activity by Fermentation of Sargassum siliquanstrum (꽈배기모자반의 발효를 통한 항염증 활성의 증진)

  • Lee, Sol-Ji;Lee, Dong-Geun;Kim, Mihyang;Kong, Chang-Suk;Yu, Ki-Hwan;Kim, Yuck-Young;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.318-324
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    • 2016
  • This study was aimed to verify anti-inflammatory activity of fermented Sargassum siliquanstrum with lactic acid bacteria. Anti-inflammatory activities were compared by measuring the amount of nitric oxide (NO) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and suppressive effect on inducible nitric oxide synthase (iNOS) expression in stably transfected RAW 264.7 cells. Inhibitory activities of NO production and iNOS expression were measured after confirmation of NO radical scavenging activities. Fermentation increased NO radical scavenging activities from 7.6% to 15.2% compared to non-fermented condition, and fermentation with Lactobacillus sp. SH-1 was the most efficient. Fermentation without algal debris showed better NO radical scavenging activities than that with debris. Fermentation with Lactobacillus sp. SH-1 also showed the highest NO production inhibitory activity (64.1%) in LPS-stimulated RAW 264.7 cells. LPS-induced iNOS expression was diminished to 28.6, 35.6, 49.4 and 58.5 at 50, 100, 500 and 1,000 μg/ml, respectively, by fermentation with Lactobacillus sp. SH-1. According to MTT assay, fermented S. siliquanstrum did not influence the cell viability at all concentrations tested, meaning no or less cytotoxicity. These results suggest that S. siliquanstrum has NO radical scavenging activity and anti-inflammatory activity. Thus biological activities of S. siliquanstrum were upgraded by fermentation, which could be used for the development of functional foods.

The Analysis of Kinetic Parameters for BNR Process Simulation in Domestic Wastewater (국내하수에 적합한 BNR 공정 시뮬레이션을 위한 최적 동력학적 계수 산출)

  • Kim, Dae-Sung;Park, Myung-Gyun;Ahn, Ho-Chul;Ahn, Won-Sik;Lee, Eui-Sin
    • Proceedings of the Korea Water Resources Association Conference
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    • 2006.05a
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    • pp.1385-1390
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    • 2006
  • 외국에서 ASM 모델의 BNR 적용 연구결과를 국내 하수에 적용하기에는 하수농도, 온도, 슬러지농도 등이 국내와는 달라 적용상 무리가 있다. 본 연구에서 BNR 시뮬레이션을 위한 입력 자료로 활용되는 인자들은 IAWPRC task group에서 제안하는 값들을 사용하되 국내 하수성상에서 필요로 하는 인자들은 직접 실험을 통하여 부분적으로 구해냄으로써 모델 시뮬레이션의 신뢰도를 높이고자 하였다. F/M비의 변화량과 1/SRT과의 관계로부터, 종속영양미생물 생산계수 $Y_H$값을 구한결과, 0.40mg VSS/mg COD였다. 이것을 ASM No.2d에 적용하기 위하여 mg cell COD formed/mg COD oxidized 단위로 환산한 결과 0.58을 나타냈다. H 하수처리장의 1차 침전지 하수를 이용하여 호기성상태에서 OUR Test를 통한 미생물에 의한 유기물 섭취시 산소섭취율 변화를 측정하였다. 호기성상태와 무산소상태에서 구한 쉽게 분해되는 용존성유기물(Ss)값을 비교해보면 각각 35.5mg/L와 39.9mg/L로 약간의 차이는 있으나 유사한 값을 보여주고 있다. 시뮬레이션을 위한 동력학적 계수 중 무산소 상태에서 종속영양미생물의 ${\mu}_{max,H}$$3.56d^{-1}$로 나타났고, 호기성상태에서는 구하면 ${\mu}_{max,H}$$4.2d^{-1}$로 산출되었다. 종속영양미생물의 사멸계수 $b_H$를 구하기 위한 실험에서 초기 OUR의 10%이내가 될 때까지 걸린 시간은 7일정도가 걸렸으며, 사멸률 $b_H$$0.043hr^{-1}$로 나타났다. 독립영양미생물의 최대비성장률 ${\mu}_{max,A}$는 최대암모니아 섭취률을 이용하여 구한 결과 $0.65d^{-1}$로 나타났다.EX>$60%{\sim}87%$가 수심 10m 이내에 분포하였고, 녹조강과 남조강이 우점하는 하절기에는 5m 이내에 주로 분포하였다. 취수탑 지점의 수심이 연중 $25{\sim}35m$를 유지하는 H호의 경우 간헐식 폭기장치를 가동하는 기간은 물론 그 외 기간에도 취수구의 심도를 표층 10m 이하로 유지 할 경우 전체 조류 유입량을 60% 이상 저감할 수 있을 것으로 조사되었다.심볼 및 색채 디자인 등의 작업이 수반되어야 하며, 이들을 고려한 인터넷용 GIS기본도를 신규 제작한다. 상습침수지구와 관련된 각종 GIS데이타와 각 기관이 보유하고 있는 공공정보 가운데 공간정보와 연계되어야 하는 자료를 인터넷 GIS를 이용하여 효율적으로 관리하기 위해서는 단계별 구축전략이 필요하다. 따라서 본 논문에서는 인터넷 GIS를 이용하여 상습침수구역관련 정보를 검색, 처리 및 분석할 수 있는 상습침수 구역 종합정보화 시스템을 구축토록 하였다.N, 항목에서 보 상류가 높게 나타났으나, 철거되지 않은 검전보나 안양대교보에 비해 그 차이가 크지 않은 것으로 나타났다.의 기상변화가 자발성 기흉 발생에 영향을 미친다고 추론할 수 있었다. 향후 본 연구에서 추론된 기상변화와 기흉 발생과의 인과관계를 확인하고 좀 더 구체화하기 위한 연구가 필요할 것이다.게 이루어질 수 있을 것으로 기대된다.는 초과수익률이 상승하지만, 이후로는 감소하므로, 반전거래전략을 활용하는 경우 주식투자기간은 24개월이하의 중단기가 적합함을 발견하였다. 이상의 행태적 측면과 투자성과측면의 실증결과를 통하여 한국주식시장에 있어서 시장수익률을 평균적으로 초과할 수 있는 거래전략은 존재하므로 이러한 전략을 개발 및 활용할 수 있으며, 특히, 한국주식시장에 적합한 거래전략은 반전거래전략이고, 이 전략의 유용성은 투자자가 설정한 투자기간보다 더욱 긴 분석기간의 주식가격정보에 의하여 최대한 발휘될 수 있음을 확인하였다.(M1), 무역적자의 폭, 산업의 생산

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Analysis of Interactions in Multiple Genes using IFSA(Independent Feature Subspace Analysis) (IFSA 알고리즘을 이용한 유전자 상호 관계 분석)

  • Kim, Hye-Jin;Choi, Seung-Jin;Bang, Sung-Yang
    • Journal of KIISE:Computer Systems and Theory
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    • v.33 no.3
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    • pp.157-165
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    • 2006
  • The change of external/internal factors of the cell rquires specific biological functions to maintain life. Such functions encourage particular genes to jnteract/regulate each other in multiple ways. Accordingly, we applied a linear decomposition model IFSA, which derives hidden variables, called the 'expression mode' that corresponds to the functions. To interpret gene interaction/regulation, we used a cross-correlation method given an expression mode. Linear decomposition models such as principal component analysis (PCA) and independent component analysis (ICA) were shown to be useful in analyzing high dimensional DNA microarray data, compared to clustering methods. These methods assume that gene expression is controlled by a linear combination of uncorrelated/indepdendent latent variables. However these methods have some difficulty in grouping similar patterns which are slightly time-delayed or asymmetric since only exactly matched Patterns are considered. In order to overcome this, we employ the (IFSA) method of [1] to locate phase- and shut-invariant features. Membership scoring functions play an important role to classify genes since linear decomposition models basically aim at data reduction not but at grouping data. We address a new function essential to the IFSA method. In this paper we stress that IFSA is useful in grouping functionally-related genes in the presence of time-shift and expression phase variance. Ultimately, we propose a new approach to investigate the multiple interaction information of genes.

The Effect of Hyperthermia Combined with Radiation on Crypts of the Mouse Jejunum (마우스공장 소낭선의 방사선 효과에 온열요법의 병용이 미치는 영향에 관한 실험적 연구)

  • Bae, Hoon-Sik;Park, Charn-Il;Kim, Jung-Jin
    • Radiation Oncology Journal
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    • v.5 no.1
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    • pp.13-21
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    • 1987
  • The effect of local hyperthermia of 41 to $43^{\circ}C$ for 30 minutes on radiosensitivity of normal tissue was studied utilizing jejunal crypt microcolony assay. Hyperthermia of this range enhanced the radiation effect and the effect was mainly additive without significant effect on the slopes of cell survival curves. At the isoeffect level of 20 microcolony formation, the thermal enhancement ratio was 1.02, 1.10 and 1.39 for $41^{\circ},\;42^{\circ}\;and\;43^{\circ}C$, respectively. The distribution of microcolony formation along the circumference of jejunum was not uniform, having more colonies around the mesenteric border, and this suggests the effect of uneven cooling by blood circulation.

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Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

Analysis of Clonorchis sinensis antigens and diagnosis of clonorchiasis using monoclonal antibodies (단세포군 항체를 이용한 간흡충 항원의 분석 및 간흡충증의 진단)

  • Yong, Tae-Sun;Im, Gyeong-Il;Jeong, Pyeong-Rim
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.293-310
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    • 1991
  • Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

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The Effect of Temperature on the Colony Development of Bombus ignitus Smith (호박벌(Bombus ignitus Smith)의 봉군발육에 미치는 영향)

  • 이상범;마영일;배태웅
    • Journal of Life Science
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    • v.9 no.4
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    • pp.395-407
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    • 1999
  • The queens of Korean native bumblebee species, Bombus ignitus were collected from the field in the spring of 1997 and reared under the various temperatures as 15$^{\circ}C$$\pm$1$^{\circ}C$, 2$0^{\circ}C$$\pm$1$^{\circ}C$, $25^{\circ}C$ $\pm$1$^{\circ}C$, 3$0^{\circ}C$$\pm$1$^{\circ}C$, 60%$\pm$5% of relative humidity and 14L : 10D, to investigate their ecological characteristics, as comparing with control(29$^{\circ}C$$\pm$1$^{\circ}C$, 60%$\pm$5% of relative humidity and red light). As a result, eggs were laid by foundation queen at 15$^{\circ}C$, but they didn't hatched out while at 2$0^{\circ}C$ eggs were laid by them and they hatched out. When a foundation queen laid eggs at $25^{\circ}C$ and they all were hatched out and normally grown. Finally they were developed to the normal colony. B. ignitus worker survived for 77days at 2$0^{\circ}C$, about 69days at $25^{\circ}C$, about 68days at 3$0^{\circ}C$ and about 63days at 29$^{\circ}C$(control). The first brood workers emerged in 25-27days from the egg which a foundation queen laid. At the various temperatures, the captive queens stand to lay eggs in about 18days at 15$^{\circ}C$, 15days at 2$0^{\circ}C$, 11days at $25^{\circ}C$, 11days at 3$0^{\circ}C$ and 4days in control. The first worker and the first drone from the egg cell of the collected queens appeared in about 27days and 72days at 2$0^{\circ}C$, 26days and 88days in $25^{\circ}C$, 24days and 65days at 3$0^{\circ}C$, 25days and 71days in control, respectively. In colony foundation, 33% of the collected queen at 2$0^{\circ}C$ and $25^{\circ}C$, 100% of the disposed queen at 3$0^{\circ}C$ and in control, and 67% of them at 3$0^{\circ}C$ and in control both produced new queens in 66days and 88days, respectively. The life span of the colony founded covered fro about 3 months at $25^{\circ}C$, 3$0^{\circ}C$ and in control. At lower temperatures, the life span of queen is shorter; 2 months at 15$^{\circ}C$ and 3 months at 2$0^{\circ}C$, respectively. A colony which normally developed, varied in size with rearing temperatures; about 20heads at 2$0^{\circ}C$, 482heads at $25^{\circ}C$, 330heads at 3$0^{\circ}C$ and 452heads in control. A foundation queen monthly oviposited 1.5egg cells at 15$^{\circ}C$, 3.0egg cells at 2 $0^{\circ}C$, 21.7egg cells at $25^{\circ}C$, 42.3egg cells at 3$0^{\circ}C$ and 47.0egg cells in control. As a colony developed in June and July, egg cells as well as daily average cells increased in number, as compared to those in May and August. Also, in June and July, interval(days) of egg laying is shorter than in May and August. Number of nectar pots during the colony development varied with rearing temperatures; 31pots at $25^{\circ}C$, 39pots at 3$0^{\circ}C$, 23pots in control. The emergence of new queen showed a distinctively different two patterns; early emerging type and late emerging type. Workers are rapidly increased from early in July to early in August at $25^{\circ}C$, but at 3$0^{\circ}C$ and in control, emergence of workers are remarkably increased from the middle of June and last until July. No new queen emerged at 15$^{\circ}C$, 2$0^{\circ}C$ and $25^{\circ}C$. New queens at 3$0^{\circ}C$ and in control emerged between late in June and early in July. Thus emergence of new queen was temperature dependent.

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