• Journal of Microbiology and Biotechnology
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• 제21권7호
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• pp.719-727
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• 2011

Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

• Mycobiology
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• 제43권2호
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• pp.137-149
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• 2015

In the search for novel potent fungi-derived bioactive compounds for bioinsecticide applications, crude ethyl acetate culture filtrate extracts from 110 mangrove fungal endophytes were screened for their toxicity. Toxicity tests of all extracts against brine shrimp (Artemia salina) larvae were performed. The extracts with the highest toxicity were further examined for insecticidal activity against Spodoptera litura larvae and acetylcholinesterase (AChE) inhibition activity. The results showed that the extracts of five isolates exhibited the highest toxicity to brine shrimp at 50% lethal concentration ($LC_{50}$) values of 7.45 to 10.24 ppm. These five fungal isolates that obtained from Rhizophora mucronata were identified based on sequence data analysis of the internal transcribed spacer region of rDNA as Aspergillus oryzae (strain BPPTCC 6036), Emericella nidulans (strains BPPTCC 6035 and BPPTCC 6038), A. tamarii (strain BPPTCC 6037), and A. versicolor (strain BPPTCC 6039). The mean percentage of S. litura larval mortality following topical application of the five extracts ranged from 16.7% to 43.3%. In the AChE inhibition assay, the inhibition rates of the five extracts ranged from 40.7% to 48.9%, while eserine (positive control) had an inhibition rate of 96.8%, at a concentration of 100 ppm. The extracts used were crude extracts, so their potential as sources of AChE inhibition compounds makes them likely candidates as neurotoxins. The high-performance liquid chromatography profiles of the five extracts differed, indicating variations in their chemical constituents. This study highlights the potential of culture filtrate ethyl acetate extracts of mangrove fungal endophytes as a source of new potential bioactive compounds for bioinsecticide applications.

• 한국수산과학회지
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• 제54권6호
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• pp.954-964
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• 2021

Mirror carp Cyprinus carpio nudus (weight 42.0±3.8 g, length 14.3±0.4 cm) were exposed to different concentrations of waterborne copper (0, 100, 200, 400, 800, and 1,600 ㎍ Cu²⁺·L^-1) at 20.3℃ for 96 h. The lethal concentration 50 of waterborne copper was 1,176.45 ㎍ Cu²⁺·L^-1. Among hematological parameters, red blood cell count was significantly decreased, whereas there were no significant changes in the hemoglobin concentration and hematocrit value. Among the inorganic plasma components, calcium was significantly decreased following copper exposure. Conversely, organic plasma components such as glucose and total protein were significantly increased. Similarly, enzymatic components, such as aspartate transaminase, alanine transaminase, and alkaline phosphatase, were also significantly increased. These findings suggest that the copper exposure is detrimental to the survival rates and physiology of C. carpio nudus.

• 분석과학
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• 제24권6호
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• pp.544-555
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• 2011

본 연구는 농약류에 대하여 구조-활성의 정량적 관계(QSAR)를 이용하여 무지개 송어(학명: Oncorhynchus mykiss)의 급성 독성을 예측-분석하는 과정을 수행하였다. 모델 구현을 위해 사용된 275종의 농약류에 대한 수중 독성(96h $LC_{50}$) 값은 DEMETRA프로젝트의 데이터를 사용하였다. 예측 모델에 사용된 2차원 분자 표현자는 PreADMET프로그램으로부터 계산을 하였고, 선형 (다중 선형 회귀 방법)모델과 비선형(서포트 벡터 머신, 인공 신경망) 학습 방법들은 실험값과 예측값의 적합도를 고려하여 최적화 되었다. 데이터 전처리 과정을 거친 뒤에, 5묶음 교차 검증과정을 포함한 모집단 기반 전진 선택법을 통해서 각 학습 방법의 최적의 표현자 집합을 결정하였다. 가장 좋은 결과는 SVM 방법 ($R^2_{CV}$=0.677, RMSECV=0.887, MSECV=0.674) 이었고, EU의 규제 기준에 따른 분류에서는 87%의 정확도를 나타내었다. MLR방법을 통해서는 무지개 송어의 급성 독성에 대하여 독성을 나타내는 농약류의 구조적 특징과 지질 층과의 상호작용을 설명할 수 있었다. 개발된 모든 모델들은 5묶음 교차 검증과 Y-scrambling test을 통해 검증되었다.

• Toxicological Research
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• 제26권4호
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• pp.293-300
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• 2010

tert-Butyl acetate (TBAc) is an organic solvent, which is commonly used in architectural coatings and industrial solvents. It has recently been exempted from the definition of a volatile organic compound (VOC) by the Air Resources Board (ARB). Since the use of TBAc as a substitute for other VOCs has increased, thus its potential risk in humans has also increased. However, its inhalation toxicity data in the literature are very limited. Hence, inhalation exposure to TBAc was carried out to investigate its toxic effects in this study. Adult male rats were exposed to TBAc for 4 h for 1 day by using a nose-only inhalation exposure chamber (low dose, $2370\;mg/m^3$ (500 ppm); high dose, $9482\;mg/m^3$ (2000 ppm)). Shamtreated control rats were exposed to clean air in the inhalation chamber for the same period. The animals were killed at 2, 7, and 15 days after exposure. At each time point, body weight measurement, bronchoalveolar lavage fluid (BALF) analysis, histopathological examination, and biochemical assay were performed. No treatment-related abnormal effects were observed in any group according to time course. Based on those findings, the median lethal concentration ($LC_{50}$) of TBAc was over $9482\;mg/m^3$ in this study. According to the MSDS, the 4 h $LC_{50}$ for TBAc for rats is over $2230\;mg/m^3$. We suggested that this value is changed and these findings may be applied in the risk assessment of TBAc which could be beneficial in a sub-acute study.

• 해양환경안전학회지
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• 제17권3호
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• pp.191-196
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• 2011

넙치 치어에 대한 나프탈렌의 급성독성을 조사하기 위하여 대조구($0{\mu}g/L$), 1000, 1800, 3200, 5600, $10000{\mu}g/L$의 6개의 나프탈렌 농도구를 설정하여 24시간 동안 노출실험을 실시하고 혈액학적 성상을 분석하였다. 넙치 치어에 대한 나프탈렌의 24h-$LC_{50}$은 $3600{\mu}g/L$를 나타냈다. 넙치 치어의 핼액학적 반응에서 헤마토크리트값은 5600, $10000{\mu}g/L$의 농도구에서 대조구와 비교하여 유의하게 증가하였고, 글루코스는 $10000{\mu}g/L$의 농도 구에서 유의하게 증가 하였다(P<0.05). 삼투질 농도는 3200, 5600, $10000{\mu}g/L$의 농도구에서 유의하게 증가하였다. 반면, 이온분석 결과 [$Na^+$], [$K^+$]은 5600 및 $10000{\mu}g/L$ 나프탈렌 농도구에서 유의하게 증가하였으나 [$Cl^-$]는 큰 차이를 보이지 않았다.

• Journal of Microbiology and Biotechnology
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• 제21권6호
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• pp.604-612
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• 2011

Beauveria bassiana SFB-205 supernatant can effectively control cotton aphid populations, which is closely associated with its chitinase activity. The present work extends to optimizing a culture medium to produce more efficacious supernatant in flask conditions, followed by scale-up in 7 L, 300 L and 1.2 KL fermentors with the parameter of chitinase. In flask conditions, a combination of soluble starch and yeast extract produced the greatest amount of chitinase (5.1 units/ml) and its supernatant had the highest aphicidal activity. An optimal quantitative combination of the two substrates, estimated by a response surface method, enabled the supernatant to have 15.7 units/ml of chitinase activity and 3.7 ml/l of median lethal concentration ($LC_{50}$) of toxicity against cotton aphid adults in laboratory conditions. In the scale-up conditions, overall supernatant had 25-28 units/ml of chitinase activity. Decrease in pH and limitation of dissolved oxygen (DO) during cultures were significantly related to the yield of chitinase. These results suggest that the substrate-dependent chitinase production can be background information for optimizing a culture medium, and pH and DO are critical factors in maximizing the production in scale-up conditions.

• Environmental Analysis Health and Toxicology
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• 제16권4호
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• pp.211-221
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• 2001

The purpose of this study was to investigate the acute (4 hours) and repeated-dose (6 hours a day, 5 days a week, 4 weeks) toxic effects of 1-hexene on Sprague-Dawley (SD) rats which were treated by inhalation. The results were as follows; 1. The median lethal concentration(LC$_{50}$) was estimated 52,694 ppm (confidence limit 95％; 49,494~55,447 ppm) in acute inhalation. Abnormal clinical signs related to the 1-Hexene were not observed with the acute inhalation dose. Cross findings of necropsy revealed on evidence of specific toxicity related to the 1-hexene. II. By repeated inhalation exposure the body weight of male were more or less reduced by the dose of 2,500 ppm and 5,000 ppm compared with control group. However there were no significant variation hematology and blood biochemistry for the exposed rats compared with the control rats. Abnormal clinical signs and gross findings of necropsy related to the 1-hexene were not shown. In conclusion when we exposed 1-hexene to SD rats for 4 weeks, 5 days per week, 6 hours per day, the Lowest observed effect level (LOEL) was over 2,500 ppm and Non observed effect level (NOEL) was below 500 ppm.

• Fisheries and Aquatic Sciences
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• 제14권3호
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• pp.205-209
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• 2011

Organic pollution causes eutrophication and dystrophication, which occur when excessive amounts of organic matter enters seawater. Eutrophication can contaminate sediment and harm aquaculture. Polychaeta species have been shown to restore eutrophic sediment. In this study, we used peptone to simulate a eutrophic environment and detect the levels at which eutrophication became toxic to the polychaete Perinereis aibuhitensis. Peptone concentrations were 0, 100, 200, and 500 mg/L. The median lethal concentrations were 950.35 mg/L at 48 h, 340.34 mg/L at 72 h, and 120.22 mg/L at 96 h, which are much higher than those of other aquatic species. Polychaeta species are highly tolerant of eutrophication. During the 15-day long-term experiment, sediment loss on ignition, as well as seawater total organic carbon and total nitrogen all decreased significantly (P<0.05). However, $NH_4^+$ concentration increased with time. Perinereis aibuhitensis slowed the increment of $NH_4^+$ but could not prevent its increase. Our results indicate that this polychaete is helpful in the recovery of seawater and sediment from eutrophication.

• Ocean and Polar Research
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• 제28권1호
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• pp.45-56
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• 2006

This study investigated primarily the toxic effects of bis(tributyltin)oxide (TBT) and DDT (Dichlorodiphenyltrichloroethane) on the mortality of adult Acartia omorii and barnacle nauplii as well as the hatching rate of A. omorii. Subsequently, compound effects of TBT and DDT on the mortality of immature copepods were tested in order to assess whether or not synergistic influence existed in the mixture of sublethal concentration of two pollutants. Mortality of adult A. omorii increased as exposure concentration of DDT increased in the range of from 0.0001 to 1ppm. Egg hatching rate of the copepod showed no distinctive difference in the range between 0.1 and 10ppm, while barnacle nauplii showed abnormal motility of their appendages in the range of 0.0001 to 1 ppm. Mortality of adult A. omorii increased as TBT concentration increased within the range of 1 and 10 ppb, whereas egg hatching rate of the copepod showed no linear response to the same exposure range. Moreover, copepod nauplii were almost motionless even though copepod eggs hatched under the exposure condition of TBT $(0{\sim}10 ppb)$ and DDT $(0{\sim}10 ppm)$, respectively, suggesting that the nauplii are hard to develop into adult stage. On the basis of the sublethal concentration less than the 24-h $LC_{50}$, 0.001 ppm (DDT) and 2 ppb (TBT) were selected to confirm the compound effects of two pollutants on the mortality of immature copepods. Mortality of immature copepods under the condition of mixture of the two pollutants was higher than that in the single exposure condition. This result seems to indicate that synergistic effects of sublethal toxicants can make a more hazardous effect on the survival of immature copepods even though the concentration of single toxicant is not lethal to copepods in the marine environment.