• Parasites, Hosts and Diseases
• /
• 제48권4호
• /
• pp.297-301
• /
• 2010

Recently, emerging waterbome protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from $10^1$ to $10^2$ oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium paNum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

• The Plant Pathology Journal
• /
• 제18권2호
• /
• pp.85-92
• /
• 2002

A new isolate of Cucumber mosaic virus (CMV), identified as Li-CMV was isolated from a diseased Korean native lily (Lithium tsingtauense Gilg). Biological and serological properties of Li-CMV were characterized, and reverse transcription-polymerase chain reaction (RT-PCR) analysis, restriction enzyme profiling of RT-PCR products, and nucleotide sequence analysis of RNA3 of the virus were performed in this study. Remarkable differences in symptoms between Li-CMV and ordinary CMV strains were found in tobacco plants and Datura stramonium. Li-CMV-infected tobacco plants (cv. Xanthi-nc and cv. Samsun) induced chlorotic ringspots on uninoculated upper leaves, and the symptom expression was delayed or faint whereas, ordinary CMV strains induced green mosaic symptoms on the plant. Systemic infections were observed on Nicotiana benthamiana with severe mosaic symptom. Restriction mapping analysis of RT-PCR products using MspI showed that Li-CMV belonged to CMV subgroup I. A full-length CDNA copy of RNA3 for the virus was amplified by RT-PCR, cloned, and its complete nucleotide sequence was determined. The RNA3 of Li-CMV was 2, 232 nucleotides long, and consisted of two open reading frames of 843 and 657 bases encoding 3a protein (movement protein) and coat protein, respectively. Results of this study indicate that Li-CMV is a novel strain and belongs to subgroup I of CMV in the genus Cucumovirus.

• The Plant Pathology Journal
• /
• 제22권1호
• /
• pp.36-45
• /
• 2006

Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

• 한국물환경학회지
• /
• 제36권3호
• /
• pp.220-228
• /
• 2020

Anaerobic ammonium oxidation (AMX) is a cost-efficient biological nitrogen removal process. The coexistence of ammonia-oxidizing bacteria (AOB) in an AMX reactor is an interesting research topic as a nitrogen-related bacterial consortium. In this study, a sequencing batch reactor for AMX (AMX-SBR) was operated with a conventional activated sludge. The AOB in an AMX bioreactor were identified and quantified using terminal restriction fragment length polymorphism (T-RFLP) and real-time qPCR. A T-RFLP assay based on the ammonia monooxygenase subunit A (amoA) gene sequences showed the presence of Nitrosomonas europaea-like AOB in the AMX-SBR. A phylogenetic tree based on the sequenced amoA gene showed that AOB were affiliated with the Nitrosomonas europaea/mobilis cluster. Throughout the enrichment period, the AOB population was stable with predominant Nitrosomonas europaea-like AOB. Two OTUs of amoA_SBR_JJY_20 (FJ577843) and amoA_SBR_JJY_9 (FJ577849) are similar to the clones from AMX-related environments. Real-time qPCR was used to quantify AOB populations over time. Interestingly, the exponential growth of AOB populations was observed during the substrate inhibition of the AMX bacteria. The specific growth rate of AOB under anaerobic conditions was only 0.111 d^-1. The growth property of Nitrosomonas europaea-like AOB may provide fundamental information about the metabolic relationship between the AMX bacteria and AOB.

• 식물병연구
• /
• 제14권3호
• /
• pp.229-232
• /
• 2008

모자이크 증상의 과꽃(Callistephus chinensis L.)으로부터 Cucumber mosaic virus (CMV)의 한 계통(Cas-CMV)를 분리하고, Fny-CMV와 As-CMV를 대조로 기주반응, dsRNA, RT-PCR 및 RFLP분석을 통하여 바이러스를 동정하였다. Cas-CMV의 특징적인 기주반응의 차이는 박과 식물에서 발현되는 강한 병정이었으며, 특히 쥬키니호박에 접종하였을 때에는 접종 15-20일 후에 심한 모자이크 증상과 함께 어린 식물이 고사되는 괴저현상을 나타냈다. DsRNA분석과 RT-PCR실험의 결과는 Cas-CMV가 서브그룹 I의 CMV에 속하는 것으로 나타났으며, 더욱이 HindIII를 이용한 RFLP 분석은 Cas-CMV가 서브그룹 IA 구분되었다.

• Journal of Preventive Medicine and Public Health
• /
• 제34권1호
• /
• pp.61-72
• /
• 2001

Objectives : To evaluate the hypothesis that increasing ambient levels of ozone or particulate matter are associated with increased emergency room visits for asthma and to quantify the strength of association, if any, between these. Methods : Daily counts of emergency room visits for asthma, air quality, and weather data were collected from hospitals with over 200 beds and from monitoring Stations in Seoul, Korea from 1994 through 1997. Daily counts of emergency mom visits for asthma attack were analyzed using a general additive Poisson model, with adjustment for the effects of secular trend, seasonal variation, Sunday and holiday, temperature, and humidly, according to levels of ozone and particulate matter. Results : The association between daily counts of emergency room visits for asthma attack and ozone levels was statistically significant in summer(from June to August), and the RR by unit inclement of 100 ppb ozone was 1.30(95% CI = $1.11\sim1.52$) without lag time. With restriction of the period from April to September in 1996, the RR was 1.37(95% CI = $1.06\sim1.76$), and from June to August in 1995, the RR was 1.62(95% CI = $1.12\sim2.35$). In the data for children$(5\sim14yr)$, the RR was 2.57(95% CI = $1.31\sim5.05$) with restriction of the period from April to September in 1997. There was no Significant association between TSP levels and asthma attacks, but a slight association was seen between PM10 levels and asthma attacks in a very restricted period. Conclusion : There was a statistically significant association between ambient levels of ozone and daily counts of emergency room visits for asthma attack. Therefore, we must make efforts to effectively minimize air pollution, in order to protect public health.

• 재활치료과학
• /
• 제7권4호
• /
• pp.19-30
• /
• 2018

목적 : 본 연구의 목적은 지역사회 거주 척수장애인의 참여 제약과 관련이 있는 요인간의 상관관계 및 그 영향을 종합적으로 분석하는 것이다. 연구방법 : 연구 대상은 지역사회에 거주하는 미국 척수 손상 협회(American Spinal Injury Assocation; ASIA) 분류법 상 손상정도(Frankel type)가 A,B,C에 해당하는 만 18세 이상 성인 척수장애인 82명이었다. 대상자들을 통하여 참여, 환경적 요인, 기능적 요인, 손상관련 요인, 심리사회적 요인, 인구사회학적 요인을 조사하였다. 참여 수준은 한국형 단축형 크레이그 핸디캡 평가(CHART-K-SF)로, 환경적 요인은 CHIEF-25 한국형으로, 기능적 요인은 척수손상환자 독립성 지수SCIM-III을 사용하였다. 손상관련 요인의 통증 및 통증관련 심리적 고통은 Pain and Distress Scale (PAD)로, 심리사회적 요인 중 자존감은 로젠버그 자아존중감 척도(RSES), 자기효능감은 일반적 자기효능감 척도(GSS)로 평가하였다. 수집된 정보는 SPSS Version 21.0을 통하여 상관관계분석, 로지스틱 회귀분석, 다중회귀분석에 사용되었다. 결과 : 참여 제약의 위험요인은 환경적 요인과 기능적 요인이었으며, 본 회귀모형의 설명력은 약 73.7%였다(R2N=.737). 환경적 장벽이 있는 것이 없는 것에 비해 약 40배 정도 참여 제약을 겪을 위험도가 높았으며, 기능적 수준이 4사분위수(상위 25%)인 것에 비해, 2사분위수일 경우 약 21배, 1사분위수(하위25%)일 경우 약 236배 정도 참여 제약을 겪을 위험도가 높아졌다. 결론 : 본 연구는 지역사회 거주 척수장애인의 참여 제약의 예측요인을 다각적인 측면으로 확인한 것에 의의가 있으며, 이를 통하여 지역사회에서 척수장애인에 대한 작업치료사의 역할을 규명한 것에 중요한 의미를 갖는다.

• Journal of Microbiology and Biotechnology
• /
• 제15권2호
• /
• pp.287-295
• /
• 2005

In order to compare bacteria] community structure and diversity in activated sludges, terminal restriction fragment length polymorphism (T-RFLP) of PCR-amplified 16s rDNAs was analyzed for 31 domestic and industrial wastewater treatment plants (WTPs). Regardless of the characteristics of the wastewaters, the bacteria] community structures of activated sludges appeared diverse and complex. In particular, activated sludges in domestic WTPs contained higher bacterial diversity than those in industrial WTPs. It was also found that terminal restriction fragment (T-RF) profiles derived from domestic WTPs were very similar with each other, although activated sludges were collected from different plants at different locations. Interestingly, activated sludges of a WTP where restaurant and toilet sewages of a company were managed showed a bacterial community structure similar to that of domestic WTPs. Activated sludges in leather industria] WTPs also showed a high similarity. However, other wastewaters possessed different bacterial communities, so that overall similarity was as low as about $30\%$. Since activated sludges from WTPs for domestic wastewaters and a company sewage appeared to hold similar bacterial communities, it was necessary to confirm if similar wastewaters induce a similar bacterial community. To answer this question, analysis of T-RFs for activated sludges, taken from another 12 domestic WTPs, was conducted by using a 6­FAM$^{TM}$-Iabeled primer and an automated DNA sequencer for higher sensitivity. Among 12 samples, it was again found that T-RF profiles of activated sludges from Yongin, Sungnam, Suwon, and Tancheon domestic WTPs in Kyonggi-do were very similar with each other. On the other hand, T-RF profiles of activated sludges from Shihwa and Ansan WTPs were quite different from each other. It was thought that this deviation was caused by wastewaters, since Ansan and Shihwa WTPs receive both domestic and industrial wastewaters. From these results, it was tentatively concluded that similar bacterial communities might be developed in activated sludges, if WTPs treat similar wastewaters.

• Asian-Australasian Journal of Animal Sciences
• /
• 제14권3호
• /
• pp.384-394
• /
• 2001

In order to prevent pollution from animal waste, P, N and pharmacological level minerals should be properly managed. Microbial phytase has been used successfully to control P excretion. Activity of natural phytase in certain plant feedstuffs is high enough to be considered in feed formulation. Nitrogen control can be achieved through amino acid supplementation and protein restriction in the diet. Supplementation with carbohydrases reduces output of excreta as well as N. Ammonia release from the manure could be reduced by using a low crude protein diet along with the supplementation with probiotics products. Excretion of minerals used at pharmacological level can be reduced by using chelated forms. Cu and Zn in the form of methionine chelate have been successfully used in the broiler and pig diets.

• 한국환경성돌연변이발암원학회지
• /
• 제16권2호
• /
• pp.77-82
• /
• 1996

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.