Salmonella spp. 11 strains에 대해 저온 열처리(50oC) 3, 6, 9분 후 MIC값을 측정하여 항생제 내성을 알아보았다. Chloramphenicol에 대해 대조군과 열처리한 strains 대부분 에서 감수성(S)이 있는 것으로 나타났고, 열처리한 strains 의 MIC값은 대조군과 비교하였을 때 유지되거나 감소하 였다. Ciprofloxacin에 대해 대조군과 열처리한 strains는 대 부분 감수성(S)이 있거나 중간(I)을 나타냈다. Tetracycline 은 모든 strains에서 감수성(S)이 있는 것으로 나타났으며, S. Gaminara BAA 711에 대해 열처리 후 MIC값이 증가 하였다. Gentamicin에 대해 대조군 strains들에서 감수성을 나타낸 strains가 3 strains, 중간을 나타낸 strains 2 strains, 내성을 가진 strains가 6 strains였으며, 이 중 S. Heidelberg ATCC 8326는 MIC값을 측정했을 때 대조군에서 MIC값 이 8 μg/mL로 MIC break point가 중간이었으나, 3분과 9 분 열처리 후 MIC값이 16 μg/mL로 증가하여 break point 가 내성을 나타냈다. 본 실험결과 Salmonella spp. 11 strains 에 대해서 저온 열처리 후 열내성 효과에 의한 항생제 내성을 알아봤을 때 ciprofloxacin에서 S. Montevideo BAA 710을 3, 6분 열처리한 경우, gentamicin에서 S. Enteritidis 109 D1을 3분 처리한 경우와 S. Heidelberg ATCC 8326 을 3, 9분 처리한 경우, tetracycline에서 S. Gaminara BAA 711을 6, 9분 처리한 경우 MIC값이 증가하였다. 후속 연 구를 통해 Salmonella spp. strains에 대해 열처리 후 열내 성 효과를 나타내는 병원성 유전자의 특성에 대한 지속적 인 연구가 필요하다.

Pharmaceutical products occurring in freshwater bodies create numerous problems for the water bodies owing to their bio-toxic nature. In order to remove such pharmaceutical pollutants, a novel Er-doped Bi4O5Br2/ g-C3N5 nanocomposite was prepared by one-pot synthesis and applied for the photocatalytic removal process. The Er ions doped on the surface of Bi4O5Br2/ g-C3N5 nanocomposite exhibited 97% degradation of tetracycline in 60 min under visible light irradiation, which is higher than pure g-C3N5 and Bi4O5Br2 photocatalysts. The improved photocatalytic properties are attributed to the outstanding visible light harvesting capacity and quick charge carrier separation efficiency which greatly reduced the recombination rate in the heterojunctions. Based on radical trapping experiments, the •O2 −, h+ and •OH radicals played a prominent role in the photodegradation reactions under visible light. Finally, the ternary Er-doped Bi4O5Br2/ g-C3N5 nanocomposite is effectively recyclable with quite a stable photocatalytic removal rate. This work enables a new perspective on the rational design of rare-earth-based nanocomposites for various pharmaceutical pollutants treatment processes.

비브리오패혈증균은 세계에서 치사율이 50%에 달하는 가장 치명적인 수인성식품매개 병원균으로 해수에서 흔히 있으며, 특히 따뜻한 계절에 발생한다. 본 연구는 제주도 의 해수, 유통 수산물, 수족관물에서 분리한 비브리오패혈 증균에 대해서 RT-PCR을 이용한 독소 유전자, Vitek을 이 용한 항생제 내성, PFGE를 이용한 유전적 특성을 조사하 였다. 총 487개의 시료를 조사한 결과 비브리오패혈증균 46주(중복 균주 포함)가 해수에서 44주, 유통수산물에서 1 주, 수족관물에서 1주 분리되었다. rtxA, viu와 같은 독소 유전자는 각각 8주(17.4%), 9주(19.6%) 검출되었고, vvhA 와 같은 독소 유전자는 모든 균주에서 검출되었다. 항생 제 내성 실험결과 cefoxitin 항상제에 대해서 100% 내성 이 나타났다. 비브리오패혈증균 46주에 대한 PFGE 분석결과 총 6유형이 100% 상동성을 보였고, 유사도는 81.3- 98.0%로 나타났다. 수산물과 수족관물에서 분리된 비브리 오패혈증균은 해수와의 상동성 결과 유사도는 불일치로 나타났고 지역과 시료 사이에는 유사성이 없었다. 독소 유 전자를 가진 비브리오패혈증균에 의한 식중독 환자가 제 주도에서 발생한 점을 고려해볼 때, 해수, 유통 수산물, 수 족관물에서 분리한 비브리오패혈증균에 대한 모니터링이 지속되어야 할 것으로 보인다.

In this study platform, electrocatalytic detection of the antibiotic chloramphenicol (CAP) in phosphate buffer (pH 7) was easily achieved using a carbon paste electrode modified with NiO nanoparticles (note NiO-CPE). The peak reduction potential of chloramphenicol on the modified electrode is at (− 0.60 V/NiO-CPE vs. Ag/AgCl), its electrochemical behavior is completely irreversible, and controlled by adsorption phenomena. An excellent electrocatalytic activity has been demonstrated by the modified elaborated electrode towards the NO2 attracting group on the side chain of chloramphenicol. The structure and chemical composition of the NiO-CPE sensor were analyzed by SEM, and the X-ray diffraction results indicated that nickel oxide microcrystals were formed on the carbon sheets. The electrochemical characteristics of the NiO-CPE sensor were examined by cyclic voltammetry and electrochemical impedance spectroscopy in comparison with the unmodified carbon. Since the DPV technique allows plotting the linearity curve between the electrocatalytic current intensity of the Chloramphenicol peak and their concentration, the proposed sensor showed a remarkable detection limit of 1.08 × 10– 8 mol/L M (S/N = 3) and a wide determination range from 100 to 0.1 μM for Chloramphenicol. The developed sensor was successfully applied in the detection of Chloramphenicol in real samples.

To increase antioxidant and antibacterial activities of seasoned soy sauce, five kinds of oriental medicinal plant(Scutellaria baicalensis (P1), Coptis japonica makino (P2), Citriunshius pericarpium (P3), Zizyphi spinosi semen (P4), Crataegus pinnatifida (P5)) and four kinds of medicinal mushrooms(Inonotus obliquus (M1), Hericium erinaceus (M2), Phellinus linteus (M3), Lentinula edodes (M4)) were added to seasoned soy sauce. Soluble solid content, pH, salinity, total polyphenol & flavonoid contents were determined. DPPH & ABTS radical scavenging activities, SOD-like activity, and antibacterial activity were analyzed. Experimental sauces showed decreased pH but significant increases of soluble solid content and salinity. Total polyphenol content was 12.76 μg GAE/g in the control. However, M1 and P1 sauce had significantly higher polyphenol contents at 352.14 and 528.25 μg GAE/g, respectively. Total flavonoids content also showed the same pattern. DPPH free radical scavenging activity was the lowest in the control at 15.75%. It was the highest at 81.80% in M1 and 68.88% in P1. ABTS free radical scavenging activity and SOD-like activity showed the same tendencies. They were higher in the experimental groups than in the control. As for the antibacterial activity analyzed by the paper-disc method, the activity increased the most in P1 and P2. In particular, P2 had the strongest antibacterial activity. Its activity against different microorganisms was in the order of Staphylococcus aureus > Bacillus cereus > Escherichia coli > Salmonella typhimurium. In conclusion, these new sauces show increased antioxidative and antioxidant activities. Therefore, they are expected to be used in various ways as a functional soy sauce.

목적 : 온도감응성 나노구조체를 제조하고 이를 적용하여, 보관시에는 항생제가 용출되지 않고, 안구착용시에 만 온도감응성으로 항생제를 용출하는 스마트 콘택트렌즈를 제조하고자 한다. 방법 : 에멀젼중합하여 p(NIPAAm)-기반의 나노구조체를 합성하였고, 이를 샌드위치 공법을 통해 콘택트렌즈 에 도입하였다. Soaking 방법을 통해 항생제인 levofloxacin(LVF)를 콘택트렌즈에 탑재하여 온도에 따라 항생제 용출 특성을 분석하였다. 결과 : sodium n-dodecyl sulfate (SDS) 마이셀 템플레이트를 활용한 에멀젼중합을 통해 20-40 nm 크기의 온도감응형 p(NIPAAm)-기반의 나노구조체를 합성하였고 이는 TEM과 입도분석기를 통해 확인하였다. 샌드위치 공법을 통해 콘택트렌즈에 나노구조체를 도입하였고, soaking 방법을 통해 항생제를 렌즈안의 나노구조체에 탑재 하였다. 25 oC와 35 oC에서 각각 항생제의 방출 특성을 분석하였다. 상온에서는 항생제를 3 ug 이내로 방출하였 지만, 35 oC에서는 2시간이내에 대부분의 항생제를 방출하였고 10 ug까지 방출하였다. 결론 : 본 연구에서는 온도감응형 나노구조체를 합성하고, 이를 콘택트렌즈에 적용 및 항생제를 탑재하여, 온 도감응형 스마트 항생제용출 콘택트렌즈를 제조하였다. 온도감응형 나노구조체는 콘택트렌즈안에서 항생제를 성 공적으로 탑재할 수 있었고, 상온에서 상당량의 항생체를 보관하고, 온도증가시 10 ug까지의 많은 양의 항생제를 방출하였다. 본 연구결과는 약물전달용 스마트 안과의료기기 및 콘택트렌즈의 개발 및 상용화에 큰 역할을 할 것 으로 기대된다.

The detection of the genome-based antibiotic resistance gene is an essential analysis process for the purpose of verifying the safety of probiotic strains, including lactic acid bacteria. In this study, 4 analysis platforms (AMRFinderPlus, staramr, rgi, ABRicate) were used for cross-comparison of 782 genomes corresponding to 19 kinds of probiotic species notified as functional foods. As a result of the analysis, the relatively fewest number of antibiotic resistance genes were detected in strains belonging to the order Lactobacillales, and antibiotic resistance genes were detected in 322 genomes used in the case of 2 types of Enterococcus genus. In addition, the presence and type of antibiotic resistance gene detection showed a lot of difference even for the same genome sequence depending on the database and analysis algorithm used by the analysis platform. These results can be confused in evaluating the potential for transmission of antibiotic resistance genes inherent in specific lactic acid bacteria and predicting potential risks that may occur in the future. Accordingly, it is judged that the antibiotic resistance gene-related analysis criteria need to be established more clearly and specifically in the safety evaluation of probiotic bacteria.

본 연구에서는 보육시설 실내공기에서 분리된 식중독 균 주의 독소 유전자 분포와 항생제 내성을 분석하여 보육시설 실내공기에 의한 식중독 발생을 사전 예방하고 식중독 발생 시 적절한 치료를 위한 기초자료를 제공하고자 하였다. 어린이집 실내공기에서 분리된 Staphylococcus aureus 16주, Bacillus cereus 37주를 실험대상으로 하였다. S. aureus와 B. cereus 독소 유전자는 PCR 방법으로 검출하였다. 항생제 감수성 실험은 Clinical and Laboratory Standard Institute의 디스크 확산법에 따라 실험하였다. S. aureus 16 균주 중 11 균주(68.6%)에서 seg와 sei 독소 유전자가 검출되었다. B. cereus 37 균주 모두에서 nheA와 nheB 독소 유전자가 검출되었다. B. cereus 독소 유전자 패턴은 총 12개로 나타났으며 nheA-nheB-nheC 독소 유전 자가 가장 중요한 패턴으로 나타났다. S. aureus 16 균주의 항생제 감수성실험 결과 ampicillin과 penicillin 항생제 에 93.8%, 87.5% 내성을 나타내었으나 methicillin resistance Staphylococcus aureus와 vacomycin resistance Staphylococcus aureus는 검출되지 않았다. B. cereus 37 균주의 항생제 감수성 실험 결과 ampicillin과 penicillin 항생제에 100% 내성을 나타냈었다. 이러한 결과를 종합하여 볼 때 보육시 설 실내공기에 오염된 S. aureus와 B. cereus에 의한 식중독을 발생을 예방하기 위하여 주기적인 환기와 공기 질 관리가 필요한 것으로 판단되었다.

본 연구는 다양한 항생제 내성을 갖는 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacininduced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM)에 대한 phage의 흡착 및 용균 특성을 평가하였다. PBST-10, PBST-13, PBST-32, PBST-35, P-22, P-22 B1 phages는 narrow host range를 보였다. 숙주인 STWT, STKCCM, STCIP에 대한 phage 의 흡착률은 각각 47-85%, 58-95%, 61-93%였지만, STCCARM 에 대한 phage의 흡착률은 14-36%의 낮은 수준을 보였다. STWT, STKCCM, STCIP, STCCARM에 대한 phage burst size는 각 각 43-350, 37-530, 66-500, 24-500 plaque-forming unit (PFU)으로 다양하게 관찰되었다. P-22 B1을 제외한 모든 phage는 배양 초기에 STCIP숙주를 효과적으로 저해하였다. 이러한 결과는 항생제 내성균을 저해하기 위해 phage control system 개발에 유용한 정보로 활용될 것이다.

As part of the research program “2018 Rapid screening and identification of freshwater microorganisms using MALDI-TOF/MS library” freshwater samples were collected from a branch of the Nakdong River. Almost 300 antibiotic-resistant bacterial strains were isolated from freshwater samples and subsequently identified by 16S rRNA gene sequencing. Seventeen strains among the isolates shared high 16S rRNA gene sequence similarity (>99.0%) with known species that were not previously recorded in Korea, and each of the isolates also formed a robust phylogenetic clade with the closest species. These species were phylogenetically diverse, belonging to four phyla, seven classes, 10 orders, and 13 genera. At the genus and class level, the previously unrecorded species belonged to Rhodovarius, Xanthobacter, and Shinella of the class Alphaproteobacteria; Ottowia, Simplicispira, and Zoogloea of Betaproteobacteria; Pseudomonas, Acinetobacter, and Shewanella of Gammaproteobacteria; Arcobacter of Epsilonproteobacteria; Sphingobacterium of Sphingobacteriia; Trichococcus of Bacilli; and Leucobacter of Actinobacteria. The previously unrecorded species were further characterized by examining their gram-staining, colony and cell morphology, biochemical properties, and phylogenetic position.

In this study, a safety evaluation was conducted to confirm if the Enterococcus faecium CKDB003 strain obtained by selection from a mixed fermentation of fruit and milk is suitable for use as a probiotic. The MIC value for the 10 antibiotics specified in the EFSA guidance was below the acceptable cut-off value. The antibiotic resistance genes aac(6')-li, eatAv, and msr(C) exist by whole genome sequencing, but are in the chromosome and not in the plasmid, thus confirming that there is no possibility of transmission to other microorganisms. It was confirmed that cytolysin (cylA, cylB, cylI, cylL-l, cylL-s, cylM, cylR1, cylR2), aggregation substance (asa1, asp1), collagen adhesion (ace), enterococcal surface protein (esp), endocarditis antigen (efaA), hyaluronidase (hyl) and gelatinase (gelE) were not present in the genome by examining the genes of factors related to virulence. Also, the biochemical analysis showed no toxic enzyme activities, and no virulence genes were detected by the PCR method. Thus, the E. faecium CKDB003 strain can be safely used as a health functional food probiotic, based on the results of the safety assessment.

본 연구는 항생제 내성 Salmonella Typhimurium CCARM 8009을 저해하기 위한 phage와 항생제 조합처리의 효과를 평가하였다. 디스크 확산법과 액체배지 희석법에 의해 phage와 항생제의 상승 저해효과를 측정하였고 배양을 통한 항생제 내성 유도를 평가하였다. Phage를 처리한 cefotaxime, chloramphenicol, ciprofloxacin, erythromycin의 디스크의 저해 구역은 각각 13.6%, 19.3%, 12.7%, 78.8% 로 증가되었다. Phage와 항생제 조합 처리에 의해 tetracycline, chloramphenicol, ciprofloxacin, erythromycin, streptomycin의 최소생육억제농도는 각각 64, 4, 0.0078, 64, 256 mg/mL으로 감소되었다. Phage와 항생제의 조합 처리는 항생제 내성 S. Typhimurium CCARM 8009을 효과적으로 저해하였다 (4 log reduction). 본 결과는 phage와 항생제의 조합처리는 항생제 내성균을 제어하기 위한 방법으로 충분히 응용가치가 높음을 보여주고 있다.

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.

Clostridium perfringens (C. perfringens) may cause diarrhea and enterotoxemia in adult and young livestock, leading to problems in the production and management of farms. Four hundred fecal samples were collected from 25 goat farms located in Gyeongsangbuk-do Province in the Republic of Korea. Sixteen C. perfringens strains were isolates from fecal samples, and the isolates were identified as type A (n=11) and type D (n=5). Additionally, α- and ε-toxin genes were detected in 16 and 5 strains by PCR, respectively, and the enterotoxin gene was presented in 2 strains. The antibiotic susceptibility test was performed using the disk diffusion method and E-test method. In the disk diffusion method, ampicillin (n=16) and chloramphenicol (n=15) were highly susceptible to 16 C. perfringens isolates. In the E-test method, ampicillin, amoxicillin, amoxicillin/clavulanic acid and meropenem were susceptible to more than 14 of 16 C. perfringens isolates. This study indicates that administration of antibiotics such as ampicillin, amoxicillin/clavulanic acid and meropenem can prevent and treat C. perfringens infections in goats.