The 20-kHz ultrasonic irradiation was applied to investigate bacterial inactivation and antibiotic susceptibility changes over time. Applied intensities of ultrasound power were varied at 27.7 W and 39.1 W by changing the amplitude 20 to 40 to three bacteria species (Escherichia coli, Enterococcus faecalis, and Staphylococcus aureus). By 15-min irradiation, E. coli, a gram-negative bacterium, showed 1.2- to 1.6-log removals, while the gram-positive bacteria, Enterococcus faecalis and Staphylococcus aureus, showed below 0.5-log removal efficiencies. Antibiotic susceptibility of penicillin-family showed a dramatic increase at E. coli, but for other antibiotic families showed no significant changes in susceptibility. Gram-positive bacteria showed no significant differences in their antibiotic susceptibilities after ultrasound irradiation. Bacterial re-survival and antibiotic susceptibility changes were measured by incubating the ultrasound-irradiated samples. After 24-hour incubation, it was found that all of three bacteria were repropagated to the 2- to 3-log greater than the initial points, and antibiotic inhibition zones were reduced compared to ones of the initial points, meaning that antibiotic resistances were also recovered. Pearson correlations between bacterial inactivation and antibiotic susceptibility showed negative relation for gram-negative bacteria, E. coli., and no significant relations between bacterial re-survival and its inhibition zone. As a preliminary study, further researches are necessary to find practical and effective conditions to achieve bacteria inactivation.

하폐수처리를 위한 분리막 생물반응기에서 막오염을 유발시키는 원인물질로 알려진 정족수감지 신호분자인 AHL과 AI-2를 동시 억제하는 연구를 하였다. AHL 분해 효소를 생산하는 BH4와 AI-2 불활성 물질을 분비하는 DKY-1를 각각 하이드로겔 담체에 고정시켜 분리막 생물반응기에 적용시킴으로써 막오염을 효과적으로 제어하고자 한다. 주기적으로 반응기의 신호분자 농도를 LC/MS/MS로 측정하였으며, 막투과 압력 증가 데이터를 바탕으로 막오염의 완화 정도를 살펴보았다. 이외에 총질소, 화학적 산소요구량, 미생물 플록 크기, 미생물 농도 등을 측정하여 반응기의 상태와 폐수의 처리효율을 비교 고찰하였다. 본 연구는 연구재단 이공분야 기초연구사업(NRF-2016R1A2B2013776)의 지원을 받아 수행되었습니다.

The lutropin/chorionicgonadotropin receptor (LHR) is a member of the rhodopsin-like subfamily of G protein-coupled receptors (GPCRs) that have been shown to mediate the internalization of its five (activation: three; inactivation: two) naturally occurring mutation. Gonadotropin receptors are members of the seven transmembrane (TM) receptor families. Several point mutations in TM II, III, V and VI have been identified in the luteinizing hormone receptor (LHR) gene, leading to constitutive activation and inactivation of the receptor. In eelLHR, we generated 3 types of constitutive activating mutations (M410T, L469R and D590Y) and 2 types of constitutive inactivating mutations (D383N and Y546F) to investigate how they work on hormone-receptor interaction. To assess the functional effects of 5 receptor mutations directly, wild-type (WT) and mutant receptors were transiently expressed in CHO-K1 cells. We evaluated the basal and cAMP stimulation by rec-LH hormone. The activity was shown to be a dose-dependent increase in cAMP production in LHR-WT expressing cells with an EC50 of 24.3 ng/ml and basal cAMP level of 2.6 nM. However, three activation mutants (D590Y, L469R and M410T) was most elevated the basal cAMP response at 12.8, 21.7 and 6.1 nM, respectively. In two inactivation mutants (D383N and Y546F) are very low in the basal cAMP activation. The EC50 was also considerably decreased to 42.3 ng/ml and 1181 ng/ml, respectively.

Black pepper (piper nigrium L.) is a spice commonly used but has a problem with microbial control, so it needs non-thermal decontamination method for product quality of dried foods. Intense pulsed light (IPL) technology is a non-thermal method for superficial decontamination of foods to inactivate pathogenic microorganisms by using high peak power and short duration pulses of a broad-spectrum (170-2600 nm) using a xenon lamp. The objective of this study was to reduce total number of bacteria in ground black pepper effectively by combined treatments of IPL and immobilized TiO2 photocatalyst. Self-designed cyclone type of pilot-scaled IPL device (> 5 kg/h) was used, which makes samples to flow cyclonically in a vacuum space longer time rather than moving vertically. Using this device alone, without TiO2 coated, 0.3-0.6 log reductions were achieved under a total energy fluence of 14.85 J/cm2 (DC voltage; 1200, 1800, and 2400 V, pulse duty; 0.5, 2.1, and 3.0 ms, treatment time; 60, 120, 180, 240, and 300 s, frequency; 2 Hz). Subsequently, TiO2-coated quartz plates with different layers between light source and samples were installed to observe the effect of photocatalyst and the efficiency of decontamination was improved slightly. However to increase the effect of the photocatalyst, several factors (TiO2 particle size, TiO2 film thickness and transparency, adhesiveness between quartz and photocatalyst, etc.) need to be concerned additionally. Nevertheless, the application of IPL treatment combined with TiO2 photocatalyst offers a potential of effective non-thermal decontamination method for dealing with powder foods in food industry.

Stream of afterglow of an atmospheric pressure plasma can conveniently be used for large scale decontamination operations. In the present study, an afterglow dielectric-barrier discharge air plasma (ADDAP) was used to inactivate Escherichia coli O157:H7 as a model microorganism for studying the plasma inactivation effect. The plasma was generated at current levels in the range of 0.4 - 0.8 A. The power consumption of ADDAP generation system ranged 169.5 - 221.9 W with respect to the current intensity range. At this current level, the temperature observed in the treatment chamber remained less than 30℃. Regarding chemical composition of ADDAP in the treatment chamber, NOx species were predominantly generated. The levels of NOx species increased as the current intensity increases and the maximum NO and NO2, concentrations noted were 6 and 4 ppm, respectively, but that of CO was less than 1 ppm level at 0.8 A. Upon treating with the ADDAP generated at 0.4 - 0.8 A for 180 min, E. coli O157:H7 showed 1.24 – 2.71 log reductions. The inactivation patterns exhibited better fit to Weibull-tail model. The comparison of delta values indicated that superior inactivation effects were observed as the current intensity increased.

This study is done to evaluate a dry sterilization method of plant pathogens on the citrus during storage. Cold plasma technique based on dielectric barrier discharge (DBD) technology was used for plasma actuating. Citrus was stored both cold (5℃, 90%RH) and ambient (18℃, 50%RH) store and analyzed for 3 weeks. Treatment (N=24) and control samples (N=48) were randomly divided for each group. The specification of the DBD plasma actuator is as follows: input voltage is 130 V, current is 0.3 A, flow rate is 15L/min, actuator operation time is 5 seconds (ON), and break time is 30 minutes (OFF). Concentration of ozone was 1.0~3.7 ppm and nitrogen dioxide was 0.5~2.5 ppm. Sterilized air was circularized by a diaphragm pump through a transparent Teflon pipe connected to the chamber and the actuator. Every 3~4 days, the selected samples were rinsed and then inoculated with potato dextrose agar (PDA) for 5 days. Cell number was counted by hemocytometer. Physical properties such as weight, color, soluble solid contents (SSC) and rigidity of peel were measured and compared during storage. Color and rigidity was measured four spots through equator of fruits.

X-chromosome inactivation is one of the most complex events observed in early embryo developments. The epigenetic changes occurred in female X-chromosome is essential to compensate dosages of X-linked genes between males and females. Because of the relevance of the epigenetic process to the normal embryo developments and stem cell studies, X-chromosome inactivation has been focused intensively for last 10 years. Initiation and regulation of the process is managed by diverse factors. Especially, proteins and non-coding RNAs encoded in X-chromosome inactivation center, and a couple of transcription factors have been reported to regulate the event. In this review, we introduce the reported factors, and how they regulate epigenetic inactivation of X-chromosomes.

There has been an accelerating increase in water reuse due to growing world population, rapid urbanization, and increasing scarcity of water resources. However, it is well recognized that water reuse practice is associated with many human health and ecological risks due to numerous chemicals and pathogenic microorganisms. Especially, the potential transmission of infectious disease by hundreds of pathogenic viruses in wastewater is one of the most serious human health risks associated with water reuse. In this study, we determined the response of different bacteriophages representing various bacteriophage groups to chlorination in real wastewater in order to identify a more reliable bacteriophage indicator system for chlorination in wastewater. Different bacteriophages were spiked into secondary effluents from wastewater plants from three different geographic areas, and then subjected to various doses of free chlorine and contact time at 5˚C in a bench-scale batch disinfection system. The inactivation of φX174 was relatively rapid and reached ∼4 log10 with a CT value of 5 mg/L*min. On the other hand, the inactivation of bacteriophage PRD1 and MS2 were much slower than the one for φX174 and only ~1 log10 inactivation was achieved by a CT value of 10 mg/L*min. Overall, the results of this study suggest that bacteriophage both MS2 and PRD1 could be a reliable indicator for human pathogenic viruses for chlorination in wastewater treatment processes and water reuse practice.

Disinfection of microorganisms using UV light is widely used in the field of water supply and wastewater treatment plant, In spite of high germicidal effect and relatively clean by-product, UV disinfection has fundamental defeat that is accumulation of fouling materials at the interface of water and lamp sleeve. Non-contact type of UV photoreactor which can avoid this fouling generation was developed and the experimental performance evaluation of the system was carried out in this study. Log inactivation rate of E. coli was selected as a disinfection index. The concentration of E. coli of second clarifier effluent was 8.2×101 - 8.2×103 colony per mL and was well inactivated by the non-contact type of UV photoreactor. Under the UV intensity condition of 2.1 - 2.5mW/cm2, E. coli removal rate was observed in the range of 54 - 95% when the HRT was increased from 10 to 52 seconds. Experimental results showed that log inactivation of E. coli was proportional to UV dosage and 200mJ/cm2 of UV dose is expected for the 2.0 log inactivation of E. coli from the second clarifier effluent. Between the two parameters of UV intensity and contact time which are consist of UV dose, UV intensity was 4 times more effective than contact time.s

비열 처리 기술인 유전체 장벽 방전 저온 플라즈마(dielectric barrier discharge cold plasma, DBD-CP)를 이용하여 양 파 분말에 접종된 Salmonella Enteritidis, Escherichia coli O157:H7, 그리고 Listeria monocytogenes의 저해 효과를 조사 하였다. DBD-CP 처리 조건으로서 DBD-CP 형성 가스는 헬륨이었고, 독립 변인은 처리 전압(4, 5, 6, 7, 8, 그리고 9 kV)과 처리 시간(5, 10, 12, 15, 그리고 20분)이었다. 미생물 저해율과 양파 분말의 표면 온도를 종속 변인으로 하여 DBD-CP 최적화 조건을 확립하였고, 이를 바탕으로 S. Enteritidis 저해율에 대한 예측 모델을 구축하였다. 또한 양파 분말의 입자 크기와 수분활성도(Aw)에 따른 S. Enteritidis 저해 효과를 알아보았다. 또다른 처리 조건으로서 헬륨-물 혼 합 가스를 형성 가스로 사용하여 9 kV에서 5분 동안 주파수(15, 25, 그리고 35 kHz)에 따른 미생물 저해율과 시료 표면 온도에 대한 영향을 관찰하였다. DBD-CP 처리에 의해 양파 분말에 접종된 S. Enteritidis, E. coli O157:H7, 그리고 L. monocytogenes는 각각 ≤2.3 ± 0.1, ≤1.4 ± 0.2, 그리고 ≤0.7 ± 1.2 log CFU/cm 2까지 저해되었고, 시료의 최고 온도는 38.5 ± 1.52℃이었다. 양파 분말에 접종된 S. Enteritidis의 저해율에 대한 DBD-CP 최적 조건은 9 kV와 20분이었고, S. Enteritidis 저해율을 가장 적절하게 예측한 모델은 Fermi’s model (R 2 =0.93)이었다. 수분활성도가 0.4, 0.8인 양파 분말의 S. Enteritidis 저해율은 각각 2.3 ± 0.1, 1.8 ± 0.1 log CFU/cm 2로 유의적인 차이가 없었고(p>0.05), 입자 크기가 0.25, 1.00 cm 2인 양파 분말의 S. Enteritidis 저해율은 각각 2.3 ± 0.1, 1.0 ± 0.5 log CFU/cm 2로 입자의 크기가 작을수록 저해율이 높았다. 헬륨-물 혼합 가스로 DBD-CP 처리시 모든 식중독균에서 주파수가 작을수록 저해율이 증가하였고, 헬륨의 단 독 처리에 비해 미생물 저해율이 유의적으로 증가하였다(p<0.05). 현 연구를 통해 DBD-CP 처리 기술은 분말 식품의 품질 보존 및 미생물 안전성을 향상시키기 위한 살균 방법으로서의 가능성을 보여주었다.

In the present study, the effects of non-thermal plasma, corona discharge plasma jet (CDPJ) against foodborne disease bacteria (Escherichia coli O157:H7 ATCC 43894, Salmonella typhimurium ATCC 13311, Staphylococcus aureus ATCC 25923) were determined, and the inactivation patterns were modeled using GinaFiT software in Microsoft Excel. Among inactivation models studied, Weibull-tail model was chosen as the best fit model based on SSE, RMSE, and r 2 values. The initial decimal reduction times (δ) of E. coli O157:H7, S. aureus, and Sal. typhimurium were 6.36, 8.31, and 12.11 s respectively. The inactivation effect increased with the current strength and was highest at span length of 25 mm. After 120 s treatment of the CDPJ at 1.50 A with span length of 25 mm, E. coli O157:H7 was reduced by 5.26 log, S. aureus by 4.21 log, Sal. typhimurium by 2.89 log.

주스의 품질악화를 초래하는 효소를 불활성화시키기 위해서 감귤 주스와 사과 주스 추출물에 초고압 처리와 열처리를 하여 각 시료의 PPO와 POD의 불활성화 정도를 확인하고 동역학 모델에 적용하여 이들의 불활성화 특성을 연구하였다. 감귤 주스와 사과 주스에서 모두 PPO가 POD보다 열과 압력에 대해서 상대적으로 내성을 갖는 것을 알 수 있었으며, 초고압 처리 과정을 거쳤을 때 보다는 열처리 과정을 거쳤을 때 PPO와 POD가 효과적으로 불활성화되는 것을 확인할 수 있었다. 초고압 기술은 처리 후에도 제품의 영양분과 색의 변화에 영향을 미치지 않기에, 최상의 상태와 품질로 제품을 제공 할 수 있는 장점을 가지고 있지만, 효소를 불활성시키기에는 부족하다는 단점을 가지고 있다. 이를 보완하여 산업적으로 이용되어지기 위해서는 영양분이 파괴되지 않는 열 처리 조건을 잡고, 효소를 효과적으로 불활성시킬 수 있는 초고압 처리 조건과 결합하여 최상의 결과를 얻을 수 있는 조건을 찾는 연구가 필요하다고 판단된다.

본 연구에서는 FCV 현탁액에 물리, 화학적 위생처리 후 복합효소처리라는 전처리과정을 적용한 뒤 real-time RTPCR법을 이용하여 살균효능을 분석하였다. RT-PCR 이전에 37oC에서 30분 동안 PK와 RNase A를 처리함으로써 UV, 열, 염소, 에탄올, 과초산계열 제품에 의해 불활성화 된 바이러스들은 음성 결과를 나타내었고, real-time RTPCR법을 통해 살균 효능을 정량분석한 결과, 복합효소처리를 했을 경우 무처리구보다 더 높은 살균 효능을 보이는 것을 확인할 수 있었다. 이로써 Nuanualsuwan S. 등11,18,29)의 선행연구에서와 같이 PK와 RNase A로 전처리하는 단계를 통하여 물리, 화학적 위생처리에 의해 손상되지 않은 바이러스가 RT-PCR 법에 의해 증폭되는 것을 방지함으로써 Real-time PCR법 에 대한 검출 감도를 높일 수 있음을 확인하였다. 또한, FCV를 검출하기 위해 사용된 RT-PCR과 real-time RT-PCR 두 방법 중에서도 real-time RT-PCR법이 가장 신속하면서도 민감도 높은 결과로 도출되었다. 따라서, 유전자 분석 이전에 복합효소처리는 물리, 화학적 위생처리에 의해 불활성화 된 바이러스의 RNA가 transcription 또는 증폭되는 것을 방지하기 위한 수단으로 real-time RT-PCR법과 결합 됨으로써 노로바이러스를 비롯한 식중독 바이러스를 검출 하는데 효과적으로 적용될 것으로 판단된다. 또한 식품현 장에서 전기영동 과정없이 신속하게 살아있는 바이러스만을 수치적으로 정량화함으로써 식품안전에도 기여할 것으 로 사료된다.

Norovirus causes acute gastroenteritis in all age groups and its food poisoning outbreaks are rapidly increasing in Korea. Reverse transcription-polymerase chain reaction (RT-PCR) is most widely used for the rapid detection of foodborne viruses due to high sensitivity. However, the false positive results of RT-PCR obtained against already inactivated viruses could be a serious drawbacks in food safety area. In this study, we investigated a method to yield true positive RT-PCR results only with alive viruses. To decompose the RNA genes from dead viruses, the enzymatic treatments composed of proteinse K and Ribonuclease A were applied to the sanitized and inactivated virus particles. Another aim of this study was to quantify the efficiencies of several major sanitizing treatments using realtime RT-PCR. Feline calicivirus (FCV) that belongs to the same Caliciviridae family with norovirus was used as a surrogate model for norovirus. The initial level of virus in control suspension was approximately 104 PFU/mL. Most of inactivated viruses treated with the enzymatic treatment for 30 min at 37oC were not detected in RT-PCR, Quantification results to verify the inactivation efficiencies of sanitizing treatments using real-time RT-PCR showed no false positive in most cases. We could successfully develope a numerical quantification process for the inactivated viruses after major sanitizing treatments using real-time RT-PCR. The results obtained in this study could provide a novel basis of rapid virus quantification in food safety area.