• 한국어병학회지
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• 제22권3호
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• pp.317-326
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• 2009

This study has shown the screening of anti-bacterial activity of three Indian medicinal plant choloroform : methanol (50:50) solvent leaf extracts (i.e. Azadirachta indica, Ocimum sanctum, and Curcuma longa) with different concentrations (10, 5, 2.5, 1.25, 0.625, 0.312, and 0.156 mg/ml) under in vitro conditions against fish pathogenic bacteria, Aeromonas hydrophila, Streptococcus iniae, Vibrio harveyi, V. anguillarum, and Edwardsiella tarda isolated from olive flounder farms, Jeju Island, South Korea. The anti-microbial activity of the A. indica and O. sanctum extracts yielded the zones of growth inhibition (ZI) was 3 and 1mm against A. hydrophila at concentration of 0.156 mg/ml when compared to that of tetracycline standard (3 mm). At highest concentration (10 mg/ml) of A. indica, O. sanctum, and C. longa, high inhibition was 9, 7, and 6 mm when compared to that of tetracycline (11 mm) against A. hydrophila. The minimum inhibitory concentration (MIC) of A. indica, O. sanctum, and C. longa at 0.156 mg/ml that yield 9, 10, and 13 CFU/ml for A. hydrophila, 16, 22, and 25 CFU/ml for S. iniae and 18, 22, and 23 CFU/ml for E. tarda compared to the tetracycline. At highest concentration (10 mg/ml) of the three extracts was better inhibiting the growth of A. hydrophila, S. iniae and E. tarda. A. indica, O. sanctum, and C. longa were determined to the potential antioxidant activityon the basis of their scavenging activity of the stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical. A. indica extract was 0.625 mg/ml which indicated that the strong anti-oxidant activity. However, O. sanctum and C. longa extracts showed weak anti-oxidant activity at this concentration. Hence, in vitro assay among the pathogens, A. hydropila is better inhibitory activity of the extracts. It is evident that the Indian medicinal plants extracts were subjected to its effectiveness against A. hydrophila, S. iniae, and E.tarda at low concentrations. The obtained results in the present study suggested that the Indian plant extracts is a prevention tools for Korean olive flounder aquaculture pathogens and its need further advance investigation.

• 한국식품조리과학회지
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• 제26권2호
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• pp.165-170
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• 2010

Although the demand of ready-to-eat (RTE) foods such as Kimbab is growing, large quantities and wide distribution of these foods is difficult due to their short shelf-life, exposed packaging with hygienic risk, and decreased quality at refrigerator temperatures. This study was undertaken to develop preservation and storage methods to extend the shelf-life of RTE rice products using microwave and packaging methods such as vacuum and modified atmosphere packages. RTE rice ball samples inoculated with Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, Staphylococcus aureus or Bacillus cereus were microwave treated for 0, 30, 60, 90 and 120 seconds. Populations of pathogens on the rice balls were significantly reduced with an increase in treatment time. There were more than 5 log reductions of all pathogens when the samples were microwave treated for 60 seconds. RTE rice balls inoculated with two pathogens (S. aureus and B. cereus) were packaged via air, vacuum, $N_2$ gas, and $CO_2$ gas following microwave treatment for 90 seconds. The initial S. aureus and B. cereus concentration before treatment was 7.60 and 6.59 log CFU/g, and these levels were reduced by 3.37 and 2.18 log CFU/g after microwave treatment. The levels of pathogens were significantly increased during storage time at room temperature. $CO_2$ packaging was the most effective at inhibiting microbial growth among the tested packaging methods. The levels of total mesophilic count, S. aureus and B. cereus after 5 days of storage were 7.7, 8.8 and 9.3 log CFU/g in air packaged samples and 2.4, 3.2 and 8.3 log CFU/g in $CO_2$ gas packaged samples, respectively. However, after 3 days of storage higher levels of B. cereus were observed in all samples, indicating that the samples were not safe to be consumed. Base on these results, microwave treatment and MAP packaging methods using $CO_2$ gas could be used as a potential method for extending the shelf-life of RTE foods.

• 한국임상수의학회지
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• 제27권1호
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• pp.1-5
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• 2010

미포자충인 Enterocytozoon bieneusi는 최근에 AIDS 환자에 기회감염 되는 병원체로 부각되었다. 이 병원체는 여러 동물에서 발견되고 있으며 인수공통 기생충으로서 특별한 관심의 대상이 되고 있다. 충남, 대전, 전북 지역의 도축장으로부터 특별한 증상을 보이지 않은 건강한 한우 총 1,729 마리의 직장변을 취하여 E. bieneusi의 감염율을 조사하였다. 조사방법은 Trichrome 염색 변법과 중합효소연쇄반응(PCR)을 이용한 분자생물학적 기법을 적용하였다. Trichrome 염색변법을 적용한 광학현미경 관찰에서 194 (11.28%) 개체에서 미포자충이 관찰되었으며, 특히 3월에 가장 높은 감염율을 보였다. 한편 분자생물학적 기법을 적용한 시험에서는 79 (4.59%) 개체에서 양성 반응을 보였으며, 이 역시 3월에 가장 높은 감염율을 나타내었다.

• 한국식품위생안전성학회지
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• 제28권4호
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• pp.312-318
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• 2013

본 연구는 인삼의 재배단계에서의 생물학적 위해요소를 조사하고 그 결과를 인삼 GAP 실천 모델의 개발을 위한 기초자료로 제공하기 위해 수행하였다. 충남 금산에 소재한 인삼 경작지 3곳에서 재배환경, 작물, 개인위생 항목에 대해 총 96점의 시료를 수집하여 위생지표세균, 병원성미생물, 그리고 곰팡이에 대해 분석하였다. 일반세균과 대장균군, 곰팡이의 오염도는 각각 1.3~6.0, 0.1~5.0 및 0.4~4.9 log CFU/g (or mL, hand, and $100cm^2$)으로 확인되었고, 대장균의 경우 C 농장의 농업용수에서 검출되었다. 병원성 미생물은 모든 시료에서 B. cereus만 0.1~4.9 log/g (or mL, hand, and $100cm^2$)범위로 검출되었으며, L. monocytogenes, E. coli O157:H7, Salmonella spp. 및 S. aureus는 검출되지 않았다. 이상의 결과 인삼 경작지 3곳은 미생물학적 위해요소에 대해서는 안전한 것으로 나타났으나, 주변 환경이나 작업자들에 의해 교차오염이 발생할 가능성은 항상 존재하므로 보다 안전성이 확보된 인삼을 재배하기 위해서 미생물학적 위해요소의 관리과 포함된 GAP 모델의 적용이 필요하다.

• 대한미생물학회지
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• 제19권1호
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• pp.11-24
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• 1984

Shigella remains to be an important enteric pathogen in this country for the present. Moreover, most of the isolates have become multiple resistant to various antibiotics which used to be drugs of choice for shigellosis. This study was made as an attempt to assess the present stage of antibiotic resistance and the incidence and transferability of R factors of Shigella. A total of one hundred and seventeen strains of Shigella isolated from patients in Seoul and provincial area between 1982 and 1983 were tested for their resistant to antimicrobial agents and transmission of R-plasmid. Antibiotic susceptibilities were determined by an agar dilution method. Muller hinton agar were used for the assay of drug resistance and tryptic soy broth were used for propagating medium for conjugation. Shigella isolated found to be one or more antibiotics were considered potential donor of R-plasmid. The following results were obtained. 1. Among 117 strains of Shigella isolated, 111 strains(94.9%) were found to be resistant to one or more drugs tested and 97.3% of these resistant strains were multiply resistant, indicating the multiply resistant strains were more than the single resistant strains. Only six strains were susceptible to all drugs tested. 2. Among 117 strains of Shigella isolated, 107 strains(91.5%) were resistant to Tetracyclin(Tc), 106 strains(90.6%) to Chloramphenicol(Cp) and Streptomycin(Sm), 97 strains(82.9%) to Ampicillin(Ap), 68 strains(58.1%) to Cephaloridine(Cr), 10 strains(8.5%) to Nalidixic acid(Na), 5 strains(4.3%) to Kanamycin(Km) and 2 strains(1.7%) to Rifampicin. No strain was resisfant to Amikacin(Ak) and Gentamicin(Gm). 3. All drug-resistant Shigella strains, except three, were multiply resistant to two or more drugs. Fifty eight strains were resistant to five drugs, followed by 26 strains resistant to dour drugs, 12 strains resistant to three drugs and 11 strains resistant to six drugs. 4. The 73% of multiply drug-resistant Shigella transferred their resistance to E. coli by conjugation and the resistance was considered to be mediated by R-plasmid. Resistance to Nalidixic acid and Rifampicin were not transferred by conjugation to recipient. As for the transferability of resistance to each seperate drug, Ap resistance was transferred with 73.2% frequence and Cm and Tc resistance were transferred with approximately 50-60% frequence whereas Sm and Cr resistance were transferred in 19.1-21.4% The other four drugs resistant failed to transfer their resistance to recipient. 5. As for the incidence and transferability of resistance to each seperate drug, the strains resistant to Tc and Cm were encountered most frequently with the rate of 91-92%, whereas transfer of Tc and Cm were low, 51-52%. The incidence of Sm resistance was very high(90.6%) but transferability of drugs resistance was much lower(25.4%). Though the incidence of Km reristance was much lower(4.3%) transferability of Km resistance was considerably higher(60%). 6. The greater the multiplicity of resistance, the greater was the likelihood that part of all of the resistance markers would be transferable.

• 한방재활의학과학회지
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• 제25권2호
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• pp.1-13
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• 2015

Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (Tris) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 추계학술대회 및 정기총회
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• pp.41-41
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• 2015

Oomycetes belong to the kingdom Straminipila, a remarkably diverse group which includes brown algae and planktonic diatoms, although they have previously been classified under the kingdom Fungi. These organisms have evolved both saprophytic and pathogenic lifestyles, and more than 60% of the known species are pathogens on plants, the majority of which are classified into the order Peronosporales (includes downy mildews, Phytophthora, and Pythium). Recent phylogenetic investigations based on DNA sequences have revealed that the diversity of oomycetes has been largely underestimated. Although morphology is the most valuable criterion for their identification and diversity, morphological species identification is time-consuming and in some groups very difficult, especially for non-taxonomists. DNA barcoding is a fast and reliable tool for identification of species, enabling us to unravel the diversity and distribution of oomycetes. Accurate species determination of plant pathogens is a prerequisite for their control and quarantine, and further for assessing their potential threat to crops. The mitochondrial cox2 gene has been widely used for identification, taxonomy and phylogeny of various oomycete groups. However, recently the cox1 gene was proposed as a DNA barcode marker instead, together with ITS rDNA. To determine which out of cox1 or cox2 is best suited as universal oomycete barcode, we compared these two genes in terms of (1) PCR efficiency for 31 representative genera, as well as for historic herbarium specimens, and (2) in terms of sequence polymorphism, intra- and interspecific divergence. The primer sets for cox2 successfully amplified all oomycete genera tested, while cox1 failed to amplify three genera. In addition, cox2 exhibited higher PCR efficiency for historic herbarium specimens, providing easier access to barcoding type material. In addition, cox2 yielded higher species identification success, with higher interspecific and lower intraspecific divergences than cox1. Therefore, cox2 is suggested as a partner DNA barcode along with ITS rDNA instead of cox1. Including the two barcoding markers, ITS rDNA and cox2 mtDNA, the multi-locus phylogenetic analyses were performed to resolve two complex clades, Bremia lactucae (lettuce downy mildew) and Peronospora effuse (spinach downy mildew) at the species level and to infer evolutionary relationships within them. The approaches discriminated all currently accepted species and revealed several previously unrecognized lineages, which are specific to a host genus or species. The sequence polymorphisms were useful to develop a real-time quantitative PCR (qPCR) assay for detection of airborne inoculum of B. lactucae and P. effusa. Specificity tests revealed that the qPCR assay is specific for detection of each species. This assay is sensitive, enabling detection of very low levels of inoculum that may be present in the field. Early detection of the pathogen, coupled with knowledge of other factors that favor downy mildew outbreaks, may enable disease forecasting for judicious timing of fungicide applications.

• 대한수의학회지
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• 제41권2호
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• pp.177-188
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• 2001

Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

• 농약과학회지
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• 제9권1호
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• pp.81-87
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• 2005

Ethaboxam[(RS)-N-(a-cyano-2-thenyl)-4-ethyl-2-(ethylamino)-1,3-thiazole-5-carboximide]은 난균강 미생물에 대하여 우수한 살균활성을 보이는 신규 살균제이다. Ethaboxam 원제와 몇 가지 ethaboxam 제형의 배추 뿌리혹병에 대한 방제효과를 조사하였다. Ethaboxam을 관주처리 하였을 때, 토양 1 L 당 8.33 mg 농도 처리구는 배추 뿌리의 혹 형성이 완전히 억제되었으며, $EC_{50}$ 값은 2.65 mg/L 토양이었다. Ethaboxam 5개 제형(10% 액상수화제, 15% 액상수화제, 2% 입제, 5% 입제, 25% 수화제) 그리고 ethaboxam과 metalaxyl 합제 제형(3%+1% 입제)의 배추 뿌리혹병 방제효과는 우수하였다. 이들 중 토양 관주처리 한 ethaboxam의 10% 및 15% 액상수화제, 토양 혼화처리 한 ethaboxam 2% 입제 제형은 다른 제형보다 배추 뿌리혹병에 대하여 더 우수한 방제효과를 보였다. 이들의 배추 뿌리혹병에 대한 $EC_{50}$ 값은 원제기준으로 토양 L 당 각각 3.72 mg(10% 액상수화제), 1.10 mg (15% 액상수화제), 4.95 mg (2% 입제) 이었다. 그러므로 실험한 ethaboxam 6개 제형 중에 15% 액상수화제가 배추 뿌리혹병에 대하여 가장 우수한 방제효과를 나타냄을 알 수 있었다. 이들 결과로부터 ethaboxam은 포장에서도 배추 뿌리혹병을 효과적으로 방제할 수 있으리라 생각되었다.

• 생명과학회지
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• 제26권6호
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• pp.737-748
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• 2016

현재까지 곤충 항균 펩티드는 1980년에 세크로피아나방(Hyalophora cecropia) 번데기의 혈림프로부터 세크로핀(cecropin)이 처음으로 정제된 이후로 150개 이상의 펩티드가 분리되어 특성들이 보고되어 왔다. 그러므로 곤충은 항균 펩티드 선발을 위한 좋은 재료로서 고려되어 왔다. 곤충 항균 펩티드는 분자량이 작으며 양전하를 띠고 다양한 길이와 서열 및 구조를 갖는 양친매성의 특징을 갖는다. 곤충 항균 펩티드는 박테리아, 진균, 기생충, 그리고 바이러스와 같은 병원체들의 침입에 대항하여 곤충의 선천성 면역체계에서 중요한 역할을 수행한다. 대부분의 곤충 항균 펩티드들은 상처가 나거나 면역화 시 지방체와 다른 특정 조직들에서 유도 합성된다. 이어서 그 항균 펩티드들은 미생물들에 대항하여 작용하기 위해 혈림프로 분비되어 나온다. 이들 펩티드들은 항암활성을 포함하여 다양한 미생물들에 대해 광범위한 항균활성을 나타낸다. 곤충 항균 펩티드는 구조 및 서열상의 특징들에 기초하여 크게 4개의 패밀리로 나누어질 수 있다. 다시 말해서 α-나선형 펩티드, 시스테인-풍부 펩티드, 프롤린-풍부 펩티드, 그리고 글리신-풍부 펩티드/단백질이 그것이다. 예를 들면, 세크로핀, 곤충 디펜신(defensin), 프롤린-풍부 펩티드, 그리고 아타신(attacin)이 일반적인 곤충 항균 펩티드들인데, 글로베린(gloverin)과 모리신(moricin)은 나비목 종들에서만 확인되어 왔다. 본 총설에서는 곤충의 항균 펩티드들에 초점을 맞추어 곤충 항균 펩티드들의 적용 가능성 및 방향과 함께 현재의 지식들과 최근의 진전된 사항들에 대하여 논의하고자 한다.