• Title/Summary/Keyword: Inoculation method

검색결과 342건 처리시간 0.034초

Detection of Clavibacter michiganensis subsp. michiganensis Assisted by Micro-Raman Spectroscopy under Laboratory Conditions

  • Perez, Moises Roberto Vallejo;Contreras, Hugo Ricardo Navarro;Herrera, Jesus A. Sosa;Avila, Jose Pablo Lara;Tobias, Hugo Magdaleno Ramirez;Martinez, Fernando Diaz-Barriga;Ramirez, Rogelio Flores;Vazquez, Angel Gabriel Rodriguez
    • The Plant Pathology Journal
    • /
    • 제34권5호
    • /
    • pp.381-392
    • /
    • 2018
  • Clavibacter michiganensis subsp. michiganesis (Cmm) is a quarantine-worthy pest in $M{\acute{e}}xico$. The implementation and validation of new technologies is necessary to reduce the time for bacterial detection in laboratory conditions and Raman spectroscopy is an ambitious technology that has all of the features needed to characterize and identify bacteria. Under controlled conditions a contagion process was induced with Cmm, the disease epidemiology was monitored. Micro-Raman spectroscopy ($532nm\;{\lambda}$ laser) technique was evaluated its performance at assisting on Cmm detection through its characteristic Raman spectrum fingerprint. Our experiment was conducted with tomato plants in a completely randomized block experimental design (13 plants ${\times}$ 4 rows). The Cmm infection was confirmed by 16S rDNA and plants showed symptoms from 48 to 72 h after inoculation, the evolution of the incidence and severity on plant population varied over time and it kept an aggregated spatial pattern. The contagion process reached 79% just 24 days after the epidemic was induced. Micro-Raman spectroscopy proved its speed, efficiency and usefulness as a non-destructive method for the preliminary detection of Cmm. Carotenoid specific bands with wavelengths at 1146 and $1510cm^{-1}$ were the distinguishable markers. Chemometric analyses showed the best performance by the implementation of PCA-LDA supervised classification algorithms applied over Raman spectrum data with 100% of performance in metrics of classifiers (sensitivity, specificity, accuracy, negative and positive predictive value) that allowed us to differentiate Cmm from other endophytic bacteria (Bacillus and Pantoea). The unsupervised KMeans algorithm showed good performance (100, 96, 98, 91 y 100%, respectively).

농촌지역 주민들의 B형 간염 예방접종 의도 및 행동 (Behavioral Intention and Behavior for Hepatitis B Vaccination in Rural Residents)

  • 오숙정;감신;한창현
    • 농촌의학ㆍ지역보건
    • /
    • 제27권1호
    • /
    • pp.125-142
    • /
    • 2002
  • This study was carried out to purpose drawing up the efficient method of rural residents' health management, analyzing factors having influences on the behavioral intention and the behavior of hepatitis B vaccination of rural residents, using the theory of planned behavior(TPB). Surveying the first questionnaire about TPB to 439 people of rural adults over 40-year old who participated to hepatitis B examination program was made from March 14 to April 23 in 2001 at two primary health care posts in a city in Gyeongsangbuk-do and the second survey for hepatitis B vaccination was made to the last subjects of 332 people(75.6%) that had been proved as subjects of inoculation against the hepatitis B as the result of examination after two months of notification. In the behavioral intention rate of hepatitis B vaccination of the subjects within a month, 100%(the top) was 45.2%, 80-90%(the middle) was 21.1% and under 70%(the low) was 33.7%. In simple analysis, vaccination intention was related with significantly health security type and economic status(p<0.05). And vaccination intention was related with significantly all variables of attitude toward behavior and subjective norm, and economic power of utilizing medical facilities and perceived power variable of perceived behavioral control(p<0.01). The vaccination rate of the subjects within a month was 31.3%. In simple analysis, the vaccination rate was higher significantly in the lower the age was, the higher the educational level was(p<0.01), the larger the family was, and subjects having transport(p<0.05). And the vaccination rate was higher in the higher the economic power was and the higher the perceived ability was. As the results of making generalized logit analysis for behavioral intention, the more positive the subjective norm was, the higher was the score of the perceived behavioral control, the intention was higher significantly in the top class on a basis of the low class of behavioral intention. In multiple logistic regression analysis for vaccination(behavior) using intention and perceived behavioral control as dependent variables, it showed that perceived behavioral control variable only influenced significantly. The higher was the score of perceived behavioral control, it showed that they got the more vaccination(p<0.01). And even in the result having analyzed, adding the significant general characteristics variables in the simple analysis to the independent variable, the higher was the score of perceived behavioral control, it showed that they got vaccination more.

  • PDF

남조류 분해세균 HY0210-AK1의 분리와 특성 및 Anabaena cylindrica 분해 활성 (Isolation and Characterization of Alga-Lytic Bacterium HY0210-AK1 and Its Degradability of Anabaena cylindrica)

  • 장은희;김정동;한명수
    • 환경생물
    • /
    • 제21권2호
    • /
    • pp.194-202
    • /
    • 2003
  • 남조류의 대발생이 자주 발생하는 석촌호수와 팔당호로부터 시료를 채취하여 남조류 분해세균의 분리를 시도하였다. MDM배지 상에서 Anabaena cylindrica를 숙주로 하는 over-layer method를 이용하여 Anabaena cylindrica lawn에서 남조류 분해 세균을 분리하여 HY0210-AK1으로 명명하였다. HY0210-AK1의 형태적, 생화학적 특성은 Rhizobium속과 유사하였으며, 16S rDNA 염기서열을 분석한 결과 Sphingobium herbicido vorans IFO 16415$^{T}$ 와 99.1%의 높은 유연관계를 보여 Sphingobium herbicidovorans HY0210-AK1로 동정하였다. 잠복기, 대수성장기, 정체기에 있는 Sphingobium herbicidovorans HY0210-AK1를 Anabaena cylindrica NIES-19와 혼합 배양하였을 때, 접종 초기에는 정체기의 Sphingobium herbicidovorans HY0210-AK1를 접종한 처리구에서 가장 좋은 살조 효과를 나타냈으나, 접종 7일 이후에는 커다란 차이를 보이지 않았다 또한,Sphingobium herbicidovorans HY0210-AK1는 $1\times 10^{8}$ CFU $ml^{-1}$ 이상의 농도로 처리하였을 때, Anabaena cylindrica NIES-19가 완전히 분해되었다. Sphingo bium herbicidovorans HY0210-AK1와 Anabaena cylin dricaNIES-19를 혼합 배양한 상등액을 여과하여 접종 하였을때는 납조류 성장 저해 효과가 나타나지 않았다. 따라서 Sphingobium herbicidovorans HY0210-AK1는 Anabaena cylindrica NIES-19를 직접 공격하여 분해하는 것으로 사료된다. Sphingobium herbicidovorans HY0210-AK1는 Microcystis aeruginosa 분해하였다..

파프리카 과실에 괴사반점을 일으키는 Pepper mild mottle virus의 병원형 (A Pathotype of Pepper mild mottle virus Causing Necrotic Spot Symptoms in Paprika Fruit)

  • 최국선;최승국;조점덕;조인숙
    • 식물병연구
    • /
    • 제19권2호
    • /
    • pp.124-127
    • /
    • 2013
  • 시설재배단지에서 파프리카의 열매에 검은 괴저 반점 증상이 발생하였다. 이 증상주를 전자현미경, 지표식물 검정 및 RT-PCR 분석 결과, 원인 바이러스로 PMMoV로 동정되었다. 신엽에서 병징은 약한 모틀 증상, 과경과 과일에 검은 반점이 보였지만, 성숙한 잎에서는 증상이 나타나지 않았다. PMMoV의 분리주들에 대한 Tobamovirus pathotype(P)의 판별 고추계통에 접종한 결과, 모두 $P_{1.2.3}$으로 확인되었다. 분리한 PMMoV를 건전 파프리카 유묘에 기계적 접종을 통하여 이 바이러스의 병원성을 증명하였다. PMMoV 분리주들의 외피단백질 유전자의 상동성은 96-99%였다. $P_{1.2}$인 PMMoV-P2 분리주의 외피단백질 아미노산 배열 139번째가 Met인 반면, 파프리카 분리주 모두 이 위치가 Asn으로 확인되었다. 본 논문은 파프리카에서 PMMoV pathotype $P_{1.2.3}$의 동정에 관한 우리나라 최초 보고이다.

Isolation and Characterization of Dextrans Produced by Leuconostoc sp. strain JYY4 from Fermented Kimchi

  • Gu, Ji-Joong;Ha, Yoo-Jin;Yoo, Sun-Kyun
    • 한국응용과학기술학회지
    • /
    • 제32권4호
    • /
    • pp.758-766
    • /
    • 2015
  • Dextran is a generic term for a bacterial exopolysaccharide synthesized from sucrose and composed of chains of D-glucose units connected by ${\alpha}$-1,6-linkages by using dextransucrases. Dextran could be used as vicosifying, stabilizing, emulsifying, gelling, bulking, dietary fiber, prebiotics, and water holding agents. We isolated new strain capable of producing dextran from Korean traditional kimchi and identified as Leuconostoc sp. strain JYY4. Batch fermentation was conducted in bioreactor with a working volume of 3 L. The media was MMY and 15% (w/v) sucrose. Mineral medium consisted of $3.0g\;KH_2PO_4$, $0.01g\;FeSO_4$, $H_2O$, $0.01g\;MnSO_4$, $4H_2O$, $0.2g\;MgSO_4\;7H_2O$, 0.01 g NaCl, $0.05g\;CaCl_2$ per 1 liter deionized water. The pH of media was initially adjusted to 6.0. The inoculation rate was 1.0% (v/v) of the working volume. Temperature was maintained at $28^{\circ}C$. The agitation rate was 100 rpm. The production pattern of dextran was associated with the cell growth. After 24 hr dextran reached its highest concentration of 59.4 g/L. The sucrose was consumed completely after 40 hr. Growth reached stationery phase when sucrose became limiting, regardless of the presence of fructose or mannitol. When the specific growth rate was 0.54 hr-1, utilization averaged 5.8 g/L-hr. The yield and productivity of dextran were 80% and 2.0 g/L-hr, respectively. Dextrans produced by were separated to two different size by an alcohol fraction method. The size of high molecular weight dextran (45% alcohol, v/v), less soluble dextran, was between MW 500,000 and 2,000,000. Soluble dextran (55% alcohol, v/v) was between 70,000 and 150,000. The molecular weight average of total dextran (70% alcohol, v/v) was between 150,000 to 500,000. The enzymatic hydrolyzates of total dextran of ATCC 13146 showed branched dextrans by Penicillium dextranase contained of glucose, isomaltose, isomaltotriose, and isomaltooligosaccharides greater than DP4 (degree of polymerization) that had branch points. Compounds greater than DP4 were branched isomaltooligosaacharides. Hydrolysates by the Lipomyces dextranase produced the same composition of oligosaccharides as those by Penicillin dextranase.

Comparison of in vitro digestibility and chemical composition among four crop straws treated by Pleurotus ostreatus

  • Nie, Haitao;Wang, Ziyu;You, Jihao;Zhu, Gang;Wang, Hengchang;Wang, Feng
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제33권1호
    • /
    • pp.24-34
    • /
    • 2020
  • Objective: The effects of Pleurotus ostreatus on the feed utilization of broad bean stalks (BBS), rape straw (RS), paddy straw (PS), and corn stalk (CS) was examined. Methods: The four roughages were co-cultured with Pleurotus ostreatus. The chemical composition; enzyme activities of laccase, carboxymethylcellulase (CMCase) and xylanase; carbohydrate and protein fractions (based on The Cornell Net Carbohydrate and Protein System [CNCPS]) were assessed at different days after inoculation (7, 14, 21, 28 d) and un-inoculated roughages (control, 0 d). The digestibility of nutrient components and the gas production of roughage with various incubation times were monitored at 0, 2, 4, 6, 9, 12, 24, 36, 48, 60, and 72 h using an in vitro ruminal fermentation method. Results: A higher CMCase activity (0.1039 U/mL) and earlier time to peak (14 d) were detected in Pleurotus ostreatus cultured with CS (p<0.05). Significantly, the incubation length-dependent responses of cumulative gas production were observed from 24 to 72 hours post fermentation (p<0.05), and these incubation length-dependent effects on cumulative gas production of PS and CS appeared earlier (24 h) for PS and CS than those (48 h) for BBS and RS (p<0.05). The fast-degradable carbohydrate (CA) content for all four roughages significantly increased over time (p<0.05). Nonetheless, increased degradation efficiency for CA treated with Pleurotus ostreatus was detected at both 21 and 28 days of incubation (p<0.05). With the exception of PS (p<0.05), there were no significant difference among the roughages (p>0.05) in slowly-degradable carbohydrate (CB2) at different incubation times (p<0.05). Conclusion: Assessment of the alterations in chemical composition, CNCPS system fractions, and the fermentation kinetics after biological pretreatment may yield a valuable database for evaluating the biological pretreatment of Pleurotus ostreatus in ruminant feed.

Correlation between Changes in Microbial/Physicochemical Properties and Persistence of Human Norovirus during Cabbage Kimchi Fermentation

  • Lee, Hee-Min;Lee, Ji-Hyun;Kim, Sung Hyun;Yoon, So-Ra;Lee, Jae Yong;Ha, Ji-Hyoung
    • Journal of Microbiology and Biotechnology
    • /
    • 제27권11호
    • /
    • pp.2019-2027
    • /
    • 2017
  • Recently, cabbage kimchi has occasionally been associated with the foodborne diseases of enteric viruses such as human norovirus (HuNoV). This study aimed to evaluate the correlation between microbial/physicochemical properties and persistence of HuNoV in experimentally contaminated cabbage kimchi fermented and stored at $4^{\circ}C$ or $10^{\circ}C$ for 28 days. Changes in organic acid content, lactic acid bacteria (LAB), acidity, pH, and salinity were analyzed. The recovery of structurally intact HuNoV was examined for up to 28 days post-inoculation, using a NoV GII.4 monoclonal antibody-conjugated immuno-magnetic separation method combined with quantitative real-time reverse transcription polymerase chain reaction. On day 0, LAB loads were $4.70log_{10}$ colony forming units/g and HuNoV GII.4 titers were $2.57log_{10}\;genomic\;copies/{\mu}l$, at both temperatures. After 28 days, intact HuNoV titers decreased to 1.58 ($4^{\circ}C$) and $1.04(10^{\circ}C)log_{10}\;genomic\;copies/{\mu}l$, whereas the LAB density increased. This correlated with a gradual increase in lactic acid and acetic acid at both temperatures. Our findings support a statistical correlation between changes in physicochemical properties and the recovery of structurally intact HuNoV GII.4. Moreover, we determined that the production of organic acid and low pH could affect HuNoV GII.4 titers in cabbage kimchi during fermentation. However, HuNoV GII.4 was not completely eliminated by microbial/physicochemical factors during fermentation, although HuNoV GII.4 was reduced. Based on this, we speculate that the persistence of HuNoV GII.4 may be affected by the continually changing conditions during kimchi fermentation.

Optimization of Indole-3-Acetic production by phosphate solubilization bacteria isolated from waste mushroom bed of Agaricus bisporus

  • Walpola, Buddhi Charana;Noh, Jae-Geun;Kim, Chan Kyem;Kyung, Ki-Cheon;Kong, Won-Sik;Yoon, Min-Ho
    • 한국버섯학회지
    • /
    • 제11권2호
    • /
    • pp.53-62
    • /
    • 2013
  • A total of 35 phosphate solubilizing bacterial strains were isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam and screened for the production of indole acetic acid (IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 ${\mu}g$ $ml^{-1}$ after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 ${\mu}g$ $ml^{-1}$) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of $30^{\circ}C$ followed by $25^{\circ}C$. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains' ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same $R_f$ value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.

랫드 발바닥 염증부위에서 관찰된 zinc함유 비만세포의 미세구조: 조직화학적 염색을 중심으로 (Ultrastructures of Zinc-containing Mast Cells Found in the Rat Hindpaw after an Inflammatory Stimuli: Zinc Selenium Autometallography)

  • 이보예;김이석;이법이;이현숙;탁계래;이영일;이정열;조승묵
    • Applied Microscopy
    • /
    • 제36권4호
    • /
    • pp.271-277
    • /
    • 2006
  • 본 연구에서는 흰쥐 발바닥에 complete Freund's adjuvant(CFA)를 투여한 후 염증부위에서 관찰되는 비만세포의 분포양상 및 미세구조를 zinc selenium autometal-lography$(AMG^{ZnSe})$로 염색한 후 광학 및 전자현미경으로 관찰하였다. 광학현미경으로 관찰된 CFA투여에 의한 염증성 반응은 피하층에 국한되어 있었으며, zinc함유 비만세포는 주로 피하조직의 혈관 주변에 분포하고 있었다. AMG 염색에 의한 흑갈색을 띠는 과립이 세포질을 채우고 있었으며, 많은 비만세포의 경우 세포막이 파괴되어 과립이 주위조직으로 퍼져 관찰되었다. 전자현미경으로 관찰된 zinc함유 비만세포는 불규칙한 외형을 띠었으며, 표면에는 작은 돌기들이 관찰되었다. AMG염색에 따른 은입자(silver grains)가 세포질내 과립에 국한되었으며, 높은 전자 밀도의 동질성 기질로 구성되어 있었으며, 경계막으로 싸여 있었다. 세포질내 세포소기관 중에서 특히 골지 복합체가 잘 발달되어 있었으며, 형질내세망과 사립체는 비교적 적게 가지고 있었다. 이들 세포의 핵은 비교적 작고 둥글고, 세포의 중심부에 위치하였으며, 이질염색질로 이루어져 있었다.

삼요소시비량과 수도생육상태가 백엽고병(벼, 흰빛잎마름병) 발병에 미치는 영향 (Effects of N,P,K fertilizer levels and growth condition on the development of Bacterial leaf blight in rice plants)

  • 김정화;조용섭
    • 한국응용곤충학회지
    • /
    • 제9권1호
    • /
    • pp.7-13
    • /
    • 1970
  • 1. 본 병에 강한 감수성품종인 금남풍을 사용하였고, Xanthomonas oyzae 6526 균주를 반합성감자배양기에 이식, $28^{\circ}C$에 3일간 사면배양한 뒤 균부유액을 만들어 상엽 상단부에 단침접종하고 14일 뒤에 확대된 병반길이를 조사하였다. 시험구설정은 3요소를 각각 무비$\cdot$표준비$\cdot$배비의 3개 수준으로 시비하여 총 27개구를 설정하고 3반복 임의배치하였다. 2. 질소질비료는 인산$\cdot$가리의 시용량에 크게 좌우되지 않았고 증시할 수록 발병이 심하였다. 3. 인산은 표준량시용하였을 때만 다소 발병억제효과의 경향이 있었으나 배비하게 되면 오히려 무비구보다도 터 발병이 촉진되었다. 4. 가리를 표준양시용하였을에 발병이 현저히 줄어들으나 시용량과 비례하지 않았고 배비하면 인산에서 처럼 다시 발병이 촉진되었다. 5. 출수시기가 늦어질 수록, 그리고 수수가 많을수록 발병이 심하였다. 6. 삼요소를 다같이 표준량시용하였을 때가 가장 발병이 적었는데, 작물의 영양조성은 비료의 종류, 시용량, 시용방법에 따라 좌우되며 발병도 기주식물의 조성과 밀접한 관계가 있으므로 시비법을 적절히 조절하면 본 병의 피해를 많이 감소시킬 수 있을 것으로 기대된다.

  • PDF