There are three kinds of Pleurotus eryngii : P. eryngii var. eryngii (P. eryngii), P. eryngii var. ferula (P. ferula : A-Wei-Mo) P. eryngii var. nebrodensis (P. nebrodensis : Bai-Ling-Gu). We bred P. ferula using Di-Mon mating and physiological assay and selected as follow (1) ‘Beesan No.1’ produced high yields, (2) ‘Beesan No.2’ was excellent morphological shape and anti-adipogenic Activit, (3) ‘Ergo’ included high ergothioneine such as anto-oxidant material, (4) ‘GW10-45’ included highest ergothioneine such as anto-oxidant material, (5) ‘Maeksong’ included high ergothioneine and effect of anti-inflammatory. We suggest that there are able to food-medicine materials.

In vivo assay of glucose detection was described using a skin tattoo film electrode (STF), and the probe was made from carbon nano tube paste modification film paper. Here in the square-wave stripping anodic working range obtained of 20-100 mgL-1 within an accumulation time of 0 seconds only in sea water electrolyte solutions of pH 7.0. The relative standard deviations of 50 mg glucose that were observed of 0.14 % (n=12), respectively, using optimum stripping accumulation of 30 sec, the low detection limit (S/N) was pegged at 15.8 mg/L. The developed results can be applied to the detect of in vivo skin sensing in real time. Which confirms the results are usable for in vitro or vivo diagnostic clinical analysis.

An investigation was conducted to evaluate the hygienic status of 53 high school foodservice systems in Gyeonggi province by using hygiene management guide checklist, ATP bioluminescence assay of food utensils were conducted during process. The 5 hygiene management guide checklist groups about personal hygiene, cooking facilities control, cross contamination control, cook and storage control, management control were checked by experts and had good grades but there were some inadequate behaviors on observation. Total cleaning levels were inadequate, including hand, rubber gloves, aprons, knives, food tray, machine and instruments. The possibility of cross contamination is also noted in handles for refrigerators, ovens, food dryers, hand washing. It was also noted that there were too much work on the nutritionist and cook, additional personnel need to be added. lack of space, deterioration of facilities were identified in some high school foodservice systems. ATP bioluminescence assay was conducted on surface of food facilities, ATP ranged 1,393±5,041.2 RLU on yellow gloves, 244±258.7 RLU on pink gloves, 3,780±11,418.6 RLU on apron, 49,056±62,831.4 RLU on refrigerator grip, 41,422±61,259.8 RLU in oven, 31,407±41,344.9 RLU on hand cleaning board.

서양종 꿀벌의 일벌의 독을 채취하여 정제한 정제봉독에 대한 세균에서의 돌연변이 유발성 검색을 위하여 S. typhimurium의 히스티딘 요구성 균주 TA100, TA1535, TA98, 그리고 TA1537의 균주와 대장균 E. coli의 트립토 판 요구성 균주인 WP2 uvrA를 이용해 복귀돌연변이시험 을 실시하였다. 정제봉독은 부형제로 사용한 멸균생리식 염수에 용해되었으며 모든 농도군의 조제물에서 침전은 관찰되지 않았으며, 정제봉독 조제물을 top agar와 혼합할 때 모든 농도군에서 혼탁이나 침전이 관찰되지 않았다. 대사활성계 적용 TA100, TA1535, TA98 균주에 대해 0, 1.5, 5, 15, 50, 150 및 500 μg/plate의 범위를 설정하고 미 적용시엔 0, 0.15, 0.5, 1.5, 5, 15 및 50 μg/plate의 범위로 설 정하였다. TA1537과 WP2 uvrA균주에 대한 농도 범위은 대사활성계 적용시엔 0, 5, 15, 50, 150, 500 및 1,500를 미 적용시엔 0, 1.5, 5, 15, 50, 150 및 500 μg/plate 범위로 설정하여 시험을 수행하였다. 그 결과 모든 시험균주에서 대사활성계 적용 여부에 상관없이 정제봉독 처리군 의 평균 집락 수는 증가를 나타내지 않았으며, 양성판정 기준을 만족시키지 못하였다. 따라서, 정제봉독은 본 시험 조건 하에 사용한 시험 균주에 복귀돌연변이를 유발하지 않는 것으로 사료되었다.

인수공통 감염증의 하나인 톡소포자충의 검출을 위해서는 대부분은 ELISA 법이 사용 되고 있으나, 충체가 사멸 된 후에도 양성반응이 나타나는 등 사용에 제한이 있다. 반면 유전자 검출법은 현재 감염상태를 확인 할 수 있기 때문에 식중독 원인조사 등에 적합하다고 판단되어 이를 활용하여 본 연구를 진행하였다. 톡소포자충의 유전정보를 통해 529 repeat region의 염기서열을 얻고, 프라이머 및 TaqMan 프로브를 설계하여 real-time PCR을 이용한 검 출법을 개발하였다. 검출한계(lower limit of detection) 및 적정곡선을 확인한 결과 10 genomic DNA copy가 검출한 계로 확인되었고, 정량을 위한 곡선은 101~106 DNA copies 까지 0.999의 R2값을 나타내었다. 개발된 검출법의 증폭효 율을 비교하기 위해 B1 gene 타겟 프라이머 세트와 타입 별 검출한계를 비교한 결과, type 1, 2, 3 톡소포자충에서 같거나 더 나은 검출한계를 보였다. 또한 식품에서 주로 분리되는 식중독 세균 14종 및 원충 3종에 대해 특이도를 비교한 결과, 모두 음성으로 나타났다. 개발된 검출법을 식육검체에 적용하였을 때 type 1, 2, 3에서 모두 원활한 검출결과를 보여 증폭방해물질이 존재하지 않은 것으로 확인되었다. 본 연구를 통해 개발된 유전자검출법은 국내 유통 중인 식육에서 인수 공통감염 원충의 하나인 톡소포 자충의 감염 여부를 확인하는 사전적 모니터링의 방법으로 활용될 예정이다.

Many turfgrass pests, Ectinohoplia rufipes, Exomala orientalis and Popillia quadriguttata, cause serious damage to grass fields in Korea. In this study, control of turfgrass pests using entomopathogenic fungi were investigated using Protaetia brevitarsis seulensis as an alternative insect. In a preliminary experiment, some Metathizium sp. showed ca. 60% virulence after 7 days exposure to the entomopathogenic fungi. Compared to the contact-exposure, overall a spray treatment showed lower control efficacy against P. brevitarsis seulensis. Furthermore, mortality of various developmental stages of P. brevitarsis seulensis was investigated under laboratory conditions. This work suggests that when treated with fungi mortality of P. brevitarsis seulensis was closely related with developmental stage. These results provide a basic information for the control of P. brevitarsis seulensis using entomopathogenic fungi

A voltammetric analysis of doxycycline was developed using DNA immobilized onto a carbon nanotube paste electrode (PE). An anodic peak current was indicated at 0.2 V (versus Ag/AgCl) in a 0.1M NH4H2PO4 electrolyte solution. The linear working range of the cyclic and square wave stripping voltammetry was obtained to 1-27 ngL-1 with an accumulation time of 800 s. Final analytical parameters were optimized to be as follows: amplitude, 0.35 V; frequency, 500 Hz; and pH, 5.43. Here detection limit was found to be 0.45 ngL-1, this result can be applied in foods systems and in the biological diagnostics

Fast, cheap and sufficient serodiagnostic tools needs to be developed for the early detectionof brucellosis. Currently the tools cannot differentiate an active infection from vaccinated, norcan it differentiate other bacterial infections with lipopolysaccharides, especially Yersiniainfections. In this study, we purified recombinant outer membrane protein 10 and 28(rOmp10,rOmp28), and a dipstick assay(indirect or sandwich) was constructed with single(rOmp10 orrOmp28) and combined rOmps(rOmp10 and rOmp28) from Brucella(B.) abortus 544 to evaluatebovine Brucella positive serum collected during the beginning of the Korean outbreak from2006 to 2015. In application with single rOmp, rOmp10(70%; indirect, 92.11%; sandwichdipstick) and rOmp28(72.5%; indirect, 86.84%; sandwich dipstick) had comparable results. Inaddition, results indicated that dipstick with combined rOmps(rOmps10 and rOmp28) weresuperior in detecting positive serum samples, at 85% indirect and 100% sandwich dipstick. Surprisingly, the results were the same in detecting negative results at 97.78% for both singleand combined indirect dipsticks. The dipstick tools with rOmp10 and rOmp28 would be usefulfor a rapid screen method for bovine brucellosis.

Loop-mediated isothermal amplification (LAMP) is a rapid, specific, cost-effective detection method by amplifying nucleic acid under isothermal conditions. In this study, we used LAMP for detection of Hamiltonella defensa that lives as a facultive endosymbiont of whitefly ‘Bemisia tabaci’. We designed the Hamiltonella-specific primers by targeting 16S ribosomal RNA gene and validated the specificity of one primer set. To find the optimum temperature for our primer set, the LAMP reaction was held at the temperature, 60℃, 62℃ and 65℃. As a result, 62℃ was the optimum reation temperature for LAMP reaction. Specificity of primer set was tested by the reaction to both Trialeurodes vaporariorum and B. tabaci. After the whole procedure, the amplicons by LAMP were visualized by adding SYBR Green to the reaction tube.

This study was conducted to identify suitable experimental conditions for the Bovine Corneal Opacity and Permeability (BCOP) assay, after which the assay was employed using 11 substances listed in the OECD test guideline (TG) 437. The opacity of the corneas was checked on days 1, 2, 3, 4, and 10 after extraction. The only day 1 showed the opacity of below 7 (limit value), which indicated the corneas have to be used in a day after extraction. The treatment time of test substances was evaluated at 3, 5, 10, 15, 20, and 30 min to determine the opacity, permeability and IVIS values. Suitable IVIS values were observed at 5, 10, and 15 min. Two culture temperature conditions, 25°C and 32°C, had similar IVIS values. The washing method suggested in the OECD TG 437 resulted in the most suitable IVIS value. Based on the established conditions, the BCOP assay was conducted using the 11 substances listed in OECD TG 437, and the sensitivity, specificity and accuracy matched those in the guidelines for all test substances. Taken together, the results of this study indicate that suitable domestic conditions for BCOP assay as an alternative eye irritation method were established. The results presented herein will be useful to future studies of other Korean alternative researches.

Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat β-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

본 연구에서는 국내 대표 식육인 소, 돼지, 닭, 오리의4종 식육과 염소, 양, 말, 칠면조의 4종 식육을 동시에 신속하게 감별할 수 있는 2 set의 multiplex PCR법을 개발하고자 미토콘드리아 16S RNA에서 종 특이부위를 선발하고 각 종에 대한 특이도를 높이기 위하여 인위적인 미스매치를 주어 프라이머를 제작한 후 8종 식육의 274개시료를 대상으로 특이도와 민감도를 조사하였다. 그 결과소, 돼지, 닭, 오리 모든 시료에서 각각 279, 94, 192, 477 bp의 증폭산물이, 말, 양, 염소, 칠면조의 모든 시료에서 각각 152 bp, 271 bp, 670 bp, 469 bp에서 뚜렷한 PCR 유전자 산물이 확인되어 모든 축종에서 100%의 특이도를 나타내어 축종별 감별력이 우수한 것으로 나타났다. 8종의 축종별로 DNA를 10 ng/μl으로 정량한 후 혼합물을 10배씩 단계 희석하여 반응여부를 조사한 결과, 소, 돼지, 오리에서는 100 fg까지, 닭에서는 1 pg까지 검출됨을 확인할수 있었다. 소, 돼지, 닭, 오리고기를 99.9%, 99%, 90%,70%, 50%, 30%, 10%, 1%, 0.1%의 비율로 혼합한 식육과 83℃ 20분, 100℃ 30분, 121℃ 10분에서 각각 열처리한 가열 혼합육에 대하여 검출한계를 조사한 결과 마지막단계의 희석 비율인 모든 혼합육의 0.1%에서 검출이 가능하였으며, 열처리 혼합육에서는 닭에서는 1% 농도에서소와 돼지의 혼합육에서 0.1% 농도에서 검출되어 민감도가 높음을 확인할 수 있었다. 본 연구에서 개발된 multiplex PCR법은 특이도 및 민감도에 있어서 국내 대표 식육을 감별하는데 있어서 유용한 것으로 평가된다.

For ex-vivo diabetic control, the voltammetric diagnosis of glucose (GU) was conducted with a modified carbon nanotube paste electrode, using handheld analytical circuits. The optimum analytical conditions were attained within the 0.5-4.0 ug/L working range and at the 0.06 ug/L detection limit, which system was interfaced to the feedback circuits and was applied to human urine for diabetic-patient diagnosis. It can be used for ex-vivo flow control analysis, vascular flow detection and other medicinal assays. The equations of the patients’ urine are y=36.65x+12.13 and R²=0.987, those of the healthy person of y= 2.5x+10.9 and R²=0.928 (patients: 118 ug/L; healthy person: 12.34 ug/L).

An investigation was conducted to evaluate the hygienic status of 33 high school foodservice systems in Yongin city by using hygiene management guide checklist, ATP bioluminescence assay and microbe inspection petrifilm (APC, coliform group, Staphylococcus aureus) of food utensils during use. The 22 hygiene management guide checklist items about facilities, personal hygiene, food control, distribution, washing and disinfection had good grade but there were some inadequate behaviors on observation. The inspection results showed their sanitary condition met the level B of the recommendation of Korea method, it means sanitary management system get settled but more practical CCP system was needed. ATP bioluminescence assay was conducted on surface of food facilities, ATP ranged 425~2,552 RLU on gloves, 541~70,251 RLU on apron, 1,596~88,490 RLU on working desk, 1,177~263,813 RLU on sterilizer grip, 715~32,814 RLU on sterilizer shelf, 114~619,725 RLU on refrigerator grip, 677~319,007 RLU on refrigerator shelf, 71~196,725 RLU on freezer grip, 1,535~233,375 RLU on freezer shelf. APC ranged 66.7±29.0 CFU on freezer grip, 102.1±35.9 CFU on refrigerator grip, 45.4±28.2 CFU on heating cabinet grip, 58.8±40.4 CFU on sterilizer grip, the number of coliform group ranged 5.6±4.9 CFU on freezer grip, 9.1±8.7 CFU on refrigerator grip, 1.2±1.1 CFU on heating cabinet grip, 4.5±4.4 CFU on sterilizer grip. S. aureus ranged 8.0±5.6, CFU on freezer grip, 12.2±9.6 CFU on refrigerator grip, 2.1±1.6 CFU on heating cabinet grip, 11.6±6.4 CFU on sterilizer grip.

The population of managed honey bees has been dramatically declining the recent past in worldwide. The one of most common disease of bees is nosemosis, the nosemosis is caused by microsporidia in the genus Nosema. Nosema apis and N. ceranae have been described as honeybee pathogens. These microsporidia are highly evoloved fungi with an obligately intracellular parasitic lifstyle. The disease causes significant detriment to honey production and results in economic losses. In our knowledge, Fumagillin is the only antibiotic approved for control of nosemosis in honey bees, however this antibiotic may have unintended effects on the honey bee host, ultimately contributing to increased prevalence and pathogenicity of Nosema. Therefore, we screened anti-Nosema substances from entomopathogenic fungal culture filtrates using in vitro polar tube germination assay. These fungal metabolites are employed as antibiotic agents. As results, Total 3 samples (23% of 13 total samples) showing the germinating inhibition against N. ceranae. This screening method may be useful for the detection of anti-Nosema substances from various samples and selected samples in this study may be a good feature to be used in the development of a new biocontrol method of nosemosis.

Entomopathogenic fungi are facultative microorganisms, dwelling in soil or infecting host insects, and some of the genera have been used as biological control agents worldwide. Collection of fungal isolates should be a platform for the development of highly effective resources, thus in this work we constructed a fungal library using a mealworm pathogenecity-based fungal collection method and further characterized some isolates with high virulence. A phylogenetic three was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, spectrum of virulence, cultural characteristics, thermo-stability of fungi, production of biologically active materials, such as enzymes. This work reports an attractive entomopathogenic fungal library including the information of effective isolates in pest management.

Entomopathogenic fungi are facultative microorganisms, dwelling in soil or infecting host insects, and some of the genera have been used as biological control agents worldwide. Collection of fungal isolates should be a platform for the development of highly effective resources, thus in this work we constructed a fungal library using a mealworm pathogenecity-based fungal collection method and further characterized some isolates with high virulence. A phylogenetic three was generated, and of the isolates 17 isolates’ biological features were characterized, such as morphology, spectrum of virulence, cultural characteristics, thermo-stability of fungi, production of biologically active materials, such as enzymes. This work reports an attractive entomopathogenic fungal library including the information of effective isolates in pest management.