The growth characterization and enzyme productivity of halotolerant microorganisms isolated from the Nakdong River estuary wetland in Korea were investigated in this study. The isolation of halotolerant microorganisms was carried out through their cultivation at 37oC for several days using a marine agar medium. After a single-colony isolation, 16 pure colonies were obtained, and phylogenetic analysis was done using 16S-based ID service in the EzBioCloud database. The isolated strains were divided into 3 phyla, 5 families, 7 genera, and 13 species. Possible industrial applications of the strains were confirmed upon testing for amylolytic, lipolytic, and proteolytic activity to confirm the possible production of hydrolytic enzymes, growth at a pH range of 5 to 9, and various salt concentrations being tested. Thirteen strains possessed at least one enzyme activity, and 5 strains possessed 2 enzyme activities. Eight strains could tolerate up to 10% NaCl concentration. This result indicated that isolated strains had shown the possible application in food and cosmetic industries. Therefore, this study would contribute to securing domestic biological resources and improving hydrolytic enzyme activity using these strains.

The two spotted mite, Tetranychus urticae Koch, is a global pest, and has developed severe resistance to several types of acaricides. This study compared the development in susceptible (S) and acequinocyl, bifenazate, pyridaben, abamectin and etoxazole-resistant (AcR, BR, PR, AbR and ER) strains of T. urticae by X-ray irradiation. When eggs were irradiated with 100 Gy, the egg hatching was completely inhibited in all strains. When nymphs were irradiated with 200 Gy, their hatching was completely inhibited in all strains. When adults were irradiated, the fecundity and egg hatching was decreased at 150 Gy and above, and hatchability of F1 generation was completely inhibited at 300 Gy in all strains. Adult longevity was slightly increase at 150 Gy and above. In addition, we performed quantitative real-time PCR on several genes. To examine the difference of all strains on radiation stress-induced gene expression, we performed quantitative RT-PCR (qPCR) of several known stress-induced genes.

목 적: 콘택트렌즈의 부작용으로 감염성 각막염이 의심되는 환자에서 분리한 균주들을 동정하고, fluoroquinolone 항생제 내성균주를 감별하여 내성기전에 관여하는 gyrA 유전자를 분석해 내성의 원인을 알아보고자 한다. 방 법: 감염성 각막염이 의심되는 기질성 침입 환자 30명을 대상으로, 각막에서 분리한 균주를 Ofloxacin(5㎍/㎖), Ciprofloxacin(5㎍/㎖), Levofloxacin(5㎍/㎖) 항생제 배지에 배양하여 16S rRNA gene PCR 방법으로 동정하였다. 디스크 확산법으로 내성균주를 감별하고, genomic DNA를 추출하여 PCR를 통 해 증폭한 후 gyrA QRDR 부위의 유전자를 분석하였다. 결 과: Ofloxacin이 첨가된 배지에서 배양된 균주는 Achromobacter xylosoxidans (N101, N102), Staphylococcus epidermidis (N105)로 동정되었고, Ciprofloxacin이 첨가된 배지에서 배양된 균주는 Achromobacter xylosoxidans (N101, N102), Staphylococcus epidermidis (N103, N104, N105, N106), Staphylococcus warneri (N107), Staphylococcus hominis (N108), Staphylococcus saprophyticus (N109), Alcaligenes sp.(N110)로 동정되었다. 항생제 디스크 확산법으로 Staphylococcus epidermidis (N105)는 Ofloxacin에 내성, Ciprofloxacin과 Levofloxacin은 중간내성으로 나타났으며, gyrA 유전자의 분 석 결과 QRDR부위에서 Ser(TCT)84 →Phe(TTT)으로 아미노산 변이가 관찰되었다. 결 론: 감염성각막염 의심환자 각막에서 그람양성균 7균주, 그람음성균 3균주가 동정되었다. fluoroquinolone 항생제 감수성 검사에서 내성균주가 나타났으며, gyrA 유전자의 codon 84 부위에서 아미 노산 변이가 나타나 내성에 영향을 미치고 있다는 것을 확인할 수 있었다. 향후 항생제의 지속적인 사용으로 많은 내성균주들이 나타날 것이라 예측되며 fluoroquinolone 항생제를 사용할 때 내성에 대한 주의가 필요 할 것으로 생각된다.

The melon and cotton aphid Aphis gossypii Glover (Hemiptera; Aphididae) is one of the most serious pests worldwide. We surveyed insecticide susceptibility in A. gossypii field populations to 12 insecticides to examine resistance ratios. The levels of insecticide resistance were extremely high, especially to neonicotinoids. One point mutation was found in the beta1 subunit loop D region of the nicotinic acetylcholine receptor (nAChR) of the imidacloprid-resistant strain. Feeding behavior analysis using an electrical penetration graph showed that sublethal doses of imidacloprid had significant effects on the duration of phloem ingestion. In addition, higher doses of imidacloprid induced contact toxicity rather than inhibition of feeding behavior. Temperature and insecticide are two important factors that affect survival, reproduction and other physiological processes of insects. To determine interactions of temperature and insecticide treatment on susceptible and imidacloprid-resistant strains of A. gossypii, adults were exposed to three temperatures (17, 22, and 28℃) or combinations of three temperatures and imidacloprid (LC20), and the expression of several genes (heat shock protein 70, cuticle protein, cytochrome P450, and elongation factor) were analyzed. Additionally, the effect of electron beam irradiation on development, reproduction, and several gene expression of imidacloprid-resistant and -susceptible A. gossypii were compared.

Temperature and insecticide are two important factors that affect survival, reproduction and other physiological processes of insects. To determine interactions of temperature and insecticide treatment on susceptible (S) and imidacloprid-resistant strains (IMI-R) of Aphis gossypii, adults were exposed to three temperatures (17, 22, and 28℃) or combinations of three temperatures (17, 22 and 28℃) and imidacloprid (LC20), and the expression of several genes (heat shock protein 70, cuticle protein, cytochrome P450, and elongation factor) were analyzed. The expression level at 17℃ of heat shock protein 70, cuticle protein, and elongation factor in S strain were up-regulated with increased time and higher than those of IMI-R strain. However, expression of cytochrome P450 was not affected by elevated temperature both S and IMI-R strain. Combined treatment of elevated temperature and imidacloprid were significantly up-regulated only cuticle protein in S strain and higher than those of IMI-R strain.

The detoxification enzymes activities were investigated to Tetranychus urticae Koch using five acaricide-resistant strains. Activities of detoxification enzymes which are glutathione S-transferase (GST), general esterases (α-naphthyl acetate and β-naphthyl acetate), and cytochrome C oxidase were determined to each resistant strain mite. Acequinocyl-resistant strain and bifenazate-resistant strain of T. urticae were showed 2.1 folds and 1.6 folds higher relative activity (RA) level of GST than susceptible strain. Other three resistant strain mites were not significant different to susceptible strain mite. General esterases and cytochrome C oxidase were not significant to all the strain of T. urticae. Acaricidal activities of acequinocylresistant strain and bifenazate-resistant strain of T. urticae showed cross-resistant both acequinocyl and bifenazate. However, other strain mites were showed susceptible acaricidal activities to two acaricides. In this result suggests that resistance of two acaricides (acequinocyl and bifenazate) might be influenced by glutathione S-tansferase activity.

The effect of electron beam irradiation on development and reproduction of imidacloprid-resistant (R strain) and -susceptible (S strain) Aphis gossypii were compared. Nymphs and adults of R and S strain were irradiated at target doses of 50, 100, 150, and 200 Gy. When nymphs were irradiated, emergence was not affected at all target dose, but number of F1 nymph was perfectly inhibited at 150 Gy in both strains. When irradiated to the adults, longevity slightly decrease at 150 Gy and above. Fecundity was strongly decreased at 100 Gy, but was not completely inhibited even at 200 Gy. Emergence of F1 nymph was decreased at 100 Gy and completely inhibited at 200 Gy. However, there were no significant differences on development and reproduction of R and S strain. We also conducted the comet assay immediately after irradiation and over the following 10 day period. Severe DNA fragmentation in A. gossypii cells was observed just after irradiation and the damage was repaired during the post-irradiation period in a time-dependent manner. These results suggest that electron beam irradiation induced abnormal development, reproduction, and DNA damage in A. gossypii, but there were no significant differences between R and S strain.

표고골목 해균인 주홍꼬리버섯을 방제하기 위해 저항성 표고균주 선발을 시도하였다. 저항성 표고균주 선발 시험에서, 사용된 77개 균주 중 67개 균주가 저항력을 가지는 것으로 확인되었다. 이들 67개 균주 중 KFRI 5를 포함한 13개 균주가 주홍꼬리버섯균의 접근을 막는데 가장 큰 영향을 미쳤고 KFRI 180을 포함한 7개 균주가 주홍꼬리버섯의 영역으로 가장 많이 침범했다. 교잡을 통해 만든 31개 균주 중 KFRI 537을 포함한 8개 균주가 주홍꼬리버섯균의 접근을 막는데 가장 큰 영향을 미쳤고 KFRI 545를 포함한 4개 균주가 주홍꼬리버섯의 영역으로 가장 많이 침범했다. 온도와 접종순서의 변화에 따른 각 균주의 저항력차이를 확인하기 위해 PDA 평판배지와 상수리나무톱밥을 넣은 시험관배지를 사용했다. PDA 평판배지 시험결과, 대부분의 처리구에서 KFRI 137은 주홍꼬리버섯에 대해 뛰어난 접근 억제력을 보였고 KFRI 180은 뛰어난 침범력을 보였다. 교잡균인 KFRI 545, 546, 547은 모균주들인KFRI 488과 405에 비해 높은 저항력을 보였다. 시험관배지 시험결과, 표고균과 주홍꼬리버섯균이 대치를 하는 시점부터 25℃에 두었을 때 모든 균주가 높은 저항력을 나타냈다. KFRI 393은 25℃에서 지속 배양할 때 다른 균주에 비해 높은 저항력을 나타냈다. 또한 모든 균주들에서, 25℃ 배양 후 두 균이 대치를 할 시기에 14℃로 옮겨 배양한 처리구는 다른 처리구들에 비해 낮은 저항력을 보였다. 이때의 저항력 정도는 각 균주 최고치의 7-20%에 불과했다. 또한 KFRI 259는 PDA 평판배지에서와 달리, 주홍꼬리버섯의 영역을 침범했다.

대부분의 식중독은 단체 급식으로부터 발생하는 경우가 많으며, 특히 위생상태와 연관되어 식중독을 야기 시키는 병인 물질 중 포도상 구균은 많은 부분을 차지 하고 있다. 따라서 본 연구에서는 서부경남지역의 5개 초등학교 급식시설에서 총 98개의 샘플 중 A급식소의 식수, D급식소의 손, E급식소의 냉장고와 앞치마에서 4개의 포도상 구균을 분리하였다. 분리된 균주들은 1개의 메티실린 저항성 혈장응고 효소 음성 황색포도상구균(Methicilline Resistant Coagulase Negative Staphylococcus aureus: MRCNS땃 깨의 메치실린 민감성 혈장응고효소 양성 황색포도상구균 (Methicilline sensitive Coagulase positive Staphylococcus aureus· MSCPS)으로 구분되었다. 한편 포도상 구균은 내열성 내독소로서, 이 중 가장 문제가 되는 내독소 B(enterotoxin B)를 검색하기 위한 PCR을 실시한 결과, A장소의 식수, D장소의 손, E 장소의 냉장고와 앞치마에서 분리된 균주로부터 477b의 생성물을 갖는 sob gene을 확인할 수 있었다. 이들 분리된 황색포도상구균에 대한 항생제 민감성 실험에서는 ampicillin과 penicillin에 대하여 전체적으로 저항성을 가졌으며, 특히 A 식수에서 분리된 균주는 옥사실린 저항성(oxacilline resistant)균주로 나타나 MRSA(methicilline resistant staphylococcus aureus)균주로 확인되었다.

온도가 살충제의 저항성과 감수성계통, 복숭아혹진덧물의 발육에 미치는 영향을 조사하기 위하여 핵형이 정상인 감수성 계통(URY-O)과 저항성 계통(ABURABI : 유일) 및 핵형이상(AI, 3 전좌)으로 저항성인 O-RY 계통을 사용하였다. 에서의 약충 기간은 저항성과 감수성 계통 간에 차이가 없었으나, 에서는 감수성 인 URY-O 계통은 8.3일간에 성층이 되었었음에 비하여 저항성인 O-RY 계통은 조사기간인 20일까지 약충상태로서 성충으로 발육하지 못하고 사망하였다. 감수성인 URY-O 계통과 저항성인 ABURABI 계통의 산자수는 에서 서로 차이가 없었으나, 저항성인 O-RY 계통의 산자 수는 에서보다 1/10밖에 자충을 낳지 못하였다. 또한 충체중은 에서 URY-O와 유일은 각각 0.22와 0.27 mg/♀이었으나, O-RY는 0.16 mg/♀로서 차이가 컸다. 가수분해활성(n mol/IS min/female)이 19인 O-RY(-)9t 88인 O-RY(+)는 , sub-strain의 산자수는 각각 3.4마리와 0.8마리로서 에스테라제 활성이 높은 O-RY(+)계통이 에스테라제 활성이 낮은 O-RY(-)계통보다 새끼수를 적게 낳았다.

본 연구에서는 선행연구를 통해 병원성 박테리아의 억제효과가 입증된 kefir 유래 유산균 L. pentosus BMSE-K006과 L. pentosus BMSE-K009의 다제내성 균주를 포함하는 Staphylococcus spp., Escherichia spp. 등 총 7주에 대한 억제효과를 확인하고자 하였 다. L. pentosus BMSE-K006과 L. pentosus BMSE-K009는 대 체로 다제내성 균주 중 그람 음성균에 대한 억제효과가 더 큰 것으 로 확인되었으며, E. coli 1507 CCARM 0236을 제외한 모든 지 시 균주에서 L. pentosus BMSE-K009보다 L. pentosus BMSEK006의 항균효과가 더 우수한 것으로 확인되었다. 본 연구는 kefir 유래 유산균의 다제내성 균주에 대한 우수한 항균효과를 입 증할 수 있는 기초자료가 될 수 있으며, 추후 probiotics 특성 및 안전성에 대한 추가적인 연구가 필요할 것으로 판단된다.

Barley Yellow Dwarf Virus (BYDV) has been a major disease causing a severe loss of yield in winter cereals worldwide. It has been recently reported that BYDV occurs frequently in wheat field and also causes serious yield reduction in Korea. This study was performed to investigate the regional distributions of BYDV strains in Korea and to identify the resistant cultivars or lines of wheat to the predominant BYDV strains, providing basic information for the breeding of BYDV-resistant wheat varieties. Using RT-PCR and EcoRI digestion methods, the regional distribution of BYDV strains in Korea from 1999 to 2000 showed that PAV strain was mainly detected about 65% (Vic-PAV 52.6% ; CN-PAV 47.4%) and MAV strain about 3%. Using ELISA test for the examination of BYDV resistance with 17 cultivars and 4 lines among Korean wheat, three cultivars, Gurumil, Topdongmil, and Olgurumil, were susceptible to BYDV and the others were resistant. In plant growth and yield component responses to BYDV infection, Gurumil showed significant difference between the uninfected and the infected, suggesting the most susceptible to BYDV among Korean wheat, but Eun-pamil and Seohae118 did no difference, an indication that they have the highest resistance.