Poultry play an important role in meeting the protein demand through the supply of chicken meat and eggs, and antimicrobial-resistant bacteria can be transmitted from poultry to humans through the food chain. In this study, 716 E. coli isolates were collected from poultry industry in Korea during the period from 2016-2018. Among the cephalosporin antimicrobial, the rate of resistance to first-cephalosporins (cefazolin and cephalexin) was more than 18.0% but secondcephalosporins (cefoxitin and cefuroxime) and four-cephalosporins (cefepime) was less than 10.0%, without any differences based on the poultry. In quinolone antimicrobial, the rate of resistance to nalidixic acid was more than 46.0% in all poultry but ciprofloxacin was more than 44.0% in broiler farm and chicken meat and less than 18.0% in layer farm and egg. In addition, the rate of resistance to ampicillin and tetracycline was more than 30.0% without any differences in poultry, but the rate of resistance to amoxicillin - clavulanate, trimethoprim - sulfamethoxazole, imipenem and gentamicin was higher in broiler farm and chicken meat than in layer farm and egg. MDR was detected in 120 (60.6%), 113 (79.6%), 80 (37.0%), and 75 (46.9%) isolates from broiler farm, chicken meat, layer farm, and egg, respectively, with an overall prevalence of 54.2% (388/716).

Royal jelly (RJ) is a well-known functional and medicinal food for human health promotion. Major royal jelly proteins (MRJPs), which are the major protein components in RJ, exhibit antimicrobial activities. However, the identities of the MRJPs of RJ responsible for its antioxidant effects have remained unclear. Here, we report that honeybee (Apis cerana) MRJP 2 (AcMRJP2) acts as an antimicrobial and antioxidant agent in RJ. Using recombinant AcMRJP2, which was produced in baculovirus-infected insect cells, we established the antimicrobial and antioxidant roles of MRJP 2. AcMRJP2 bound to the surfaces of bacteria, fungi, and yeast, which then induced structural damage in the microbial cell walls and led to a broad spectrum of antimicrobial activities. AcMRJP2 protected mammalian and insect cells via the direct shielding of the cell against oxidative stress, which led to reduced levels of caspase-3 activity and oxidative stress-induced cell apoptosis, followed by increased cell viability. Moreover, AcMRJP2 exhibited DNA protection activity against reactive oxygen species (ROS). Our data indicate that AcMRJP2 could play a crucial role as an antimicrobial agent and antioxidant in RJ, suggesting that MRJP 2 is a component responsible for the antimicrobial and antioxidant activities of RJ.

Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.

It has been well known that IKK-β, -ε and –γ play a pivotal role in IMD pathway. In this study, TmIKK-ε was identified and their functions in countering pathogenic infections were investigated. We identified TmIKK-ε gene which including 2,196 bp nucleotides (encoding 731 amino acid residues). Domain analysis of TmIKK-ε indicates that there is one Serine/Threonine protein kinases catalytic domain. TmIKK-ε gene was highly expressed in 2 day-old pupal stage and the expression was gradually decreased until 1 day-old adults. Then the expression was slightly increased until 4 day-old adult stage. Tissue specific expression of TmIKK-ε mRNA was high in the gut, integuments and hemocytes in last instar larvae, and fat body, Malpighian tubules and testis in 5-daysold adult. In hemocytes, TmIKK-ε was drastically induced by E. coli injection after 3 h and by S. aureus at 3 and 12 h-post injection. In gut, expression level of TmIKK-ε was high at 6 h-post injection of microbial injection. Expression of TmIKK-ε in fat body was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-ε, gene specific RNAi and mortality assay were performed. TmIKK-ε RNAi caused increased larval mortality against E. coli, not S. aureus and C. albicans. Finally, to investigate the induction patterns of Tenebrio fourteen AMP genes in response TmIKK-ε RNAi, three microorganisms were treated into TmIKK-ε-silenced T. molitor larvae. Nine out of fourteen AMP genes were not induced by microbial challenge in TmIKK-β dsRNA-injected group. Taken together, our results indicate that TmIKK-ε may regulates nine antimicrobial peptide genes in response to microbial challenge in T. molitor fat body.

IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.

목적 : Coagulase-negative Staphylococci 분리 균주에 대한 quinolone 계열 및 그람양성균에 효과적인 vancomycin과 erythromycin 항생제 감수성 정도를 알아보고 DNA gyrase 유전자를 분석하고자 한다. 방법 : 세균성 각막염이 의심되는 사람의 각막에서 검출한 Coagulase-negative Staphylococci 6 균주를 대상으로 ofloxacin, ciprofloxacin, levofloxacin, vancomycin, erythromycin 항생제 감수성을 실시한 후 각 균주들의 genomic DNA를 추출하여 PCR 증폭한 후 gyraseA 유전자의 DNA를 분석하였다. 결과 : 항생제 감수성 검사에서 S. epidermidis (SE-2, SE-3), S. hominis (SH-1) 그리고 S. saprophyticus (SS-1) 균주에서는 ofloxacin, ciprofloxacin, levofloxacin, vancomycin 감수성, erythromycin 내성으로 나타났으며, 나머지 균주에서는 모두 감수성으로 나타났다. gyraseA 유전자 분석에서는 S. epidermidis (SE-1) 균주에서 Met75→Thr, Met93→Val 으로 아미노산 이중 mutation이 관찰되었으며, S. warneri (SW-1)와 S. saprophyticus (SS-1) 균주에서는 다수의 silent mutation이 관찰되었다. 결론 : Coagulase-negative Staphylococci 분리균주에서 erythromycin 항생제의 내성이 있다는 것을 확인 할 수 있었으며, gyraseA 유전자 분석결과 아미노산 이중 mutation 및 silent mutation이 관찰되었다. 따라서 지금은 내성이 아니지만 silent mutation이 언제든지 내성을 일으킬 수 있을 것으로 생각된다.

Enterococcus faecalis is a major causative agent of endodontic treatment failure. The purpose of this study was to investigate bactericidal effects of ethanol extract of Garcinia mangostana L. (mangosteen extract) on five strains of E. faecalis that were isolated from human oral cavities. The bactericidal effects of mangosteen extract were assessed by measurement of minimum bactericidal concentration (MBC) value. The cytotoxicity of mangosteen extract on immortalized human gingival fibroblasts, hTERT-hNOF, was determined based on cell counting method. The data revealed the MBC value of mangosteen extract against the E. faecalis strains was 4 ㎍/ml. Additionally, the cell viability of mangosteen extract on hTERT-hNOF was 83.7-89.1% at the 1 to 16 ㎍/ml. These findings indicated that mangosteen extract could be used as a root canal cleaner during management of endodontic treatment failure caused by E. faecalis.

Polyphenon 60 refers to the mixture of catechins present in green tea. The aim of this study was to investigate the antimicrobial activities of polyphenon 60 against 4 strains of Streptococcus mutans and 2 strains of Streptococcus sorbrinus, which are the major causative bacteria of dental caries. The minimum bactericidal concentration (MBC) values of polyphenon 60 for S. mutans and S. sobrinus were determined and the effect of biofilm formation inhibition of that was evaluated. The MBC value of polyphenon 60 against the bacterial strains was 2.5 mg/ml except for one particular strain, S. mutans KCOM 1128 for which the value was 1.25 mg/ml. The results of biofilm formation inhibition assay revealed that polyphenon 60 inhibited biofilm formation more than 90% at a concentration of 2.5 mg/ml. It was apparent that polyphenon exhibited biofilm formation inhibition activity along with bactericidal effect against S. mutans and S. sobrinus. Therefore, it is proposed that polyphenon 60 as one of the components of bactericidal agents could be useful in developing oral hygiene products, toothpaste or gargling solution.

본 연구는 제주산 자생 로즈마리 에센셜 오일의 항염 및 피부상재균에 대한 항균 활성을 확인하기 위한 것이다. 로즈마리 에센셜 오일은 물 증류법으로 추출하였다. 로즈마리 에센셜 오일의 항염 효능을 확인하기 위하여 RAW 264.7 세포에서 LPS에 의해 유도된 NO와 PGE2의 생성을 농도 의존적으로 억제하는 것을 확인하였다. Western blot 실험을 통해 이들을 생합성하는 효소인 iNOS, COX-2 단백질의 발현이 농도 의존적으로 감소하는 것을 확인하였다. 또한 전염증성 cytokine인 TNF-⍺와 IL-6의 생성 억제 효능을 확인하였다. 항생제 내성균주 각 2종을 포함한 S. epidermidis 3종, P. acnes 3종에 대한 항균 활성을 실험한 결과, paper disc법에서 저해환이 관찰되었고, MIC, MBC 실험을 통하여 균생육 저해와 사멸를 확인하였다. 위의 실험 결과로부터 로즈마리 에센셜 오일의 항염증, 항균 효능을 확인 하였으며 향후 화장품 및 스킨케어 소재로서 활용 가능성을 확인하였다.